chapter ii review of literature -...

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CHAPTER II

REVIEW OF LITERATURE

In recent years, considerable attention has been focused on evaluation of the effects plants on

fertility enhancing, anti-fertility or any other reproductive problems throughout the world. Plants

have been always regarded as safe medicine for any kind of diseases all over the world.

Problems of reproductive system and fertility are a big issue today. Data suggested that the

fertility rate in all the countries has been declining in a progressive rate. The reason for decline of

fertility has been suspected due to many causes which has been discussed in the Chapter-I.

Scientists now trying to invent drugs from plant origin to overcome fertility problems as these

drugs are believed to more safe than any other drugs. Many workers reported different plants to

enhance fertility in recent years in both male and female individuals.

2.1: EVALUATION OF FERTILITY ENHANCING PROPERTIES OF MEDICINAL

PLANTS:

Plants having fertility enhancing property may act as an aphrodisiac, may enhance reproductive

performances like mating behavior, mounting behavior etc., or they may help in the treatment of

different reproductive diseases. Besides these plants having fertility enhancing nature are

reported to increase sperm quality such as number, motility along with enhanced functioning of

the reproductive organs in male. It has also been reported that plants which enhance fertility in

male increase serum sex hormones in male. In female fertility enhancing property of a plant has

been established on the parameters like estrogenicity, implantation numbers, litter size, post

implantation loss, serum sex hormonal levels etc.

Aphrodisiac property of the plant, Argyreia nervosa was reported by A. Subramoniam and his

colleagues in 2007. They used root, flower and the stem of the plant to evaluate the aphrodisiac

nature of the plant on male mice. They found that the treated mice showed increased mounting

behavior and mating performance compared to the control mice. Aphrodisiac property of

Eurycoma longifolia Jack was studied by Ang et al., 2000. Butanol, methanol, water and

chloroform extracts of the roots of E. longifolia Jack was orally administered to male rats for 10

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days. Results of their study showed a dose dependent increase in the sexual performance in

mounting, intromission and ejaculation was observed in the treated groups when compared to the

control. Fertility enhancing effects of Lophira lanceolata was tested on rats for testicular and

epididymal weights, epididymal sperm number, sperm motility and morphology by Etuk and

Muhammad, 2009. They observed that the aqueous stem bark extract increased testicular and

epididymal weights as well as sperm number. But it was reported that, there was no alteration in

sperm motility and morphology in the treated animals when compared to the control animals.

Mucuna pruriens L. is a frequently recorded herb for enhancing fertility in male. It increases

sperm number, sperm quality, sperm morphology and enhances male fertility (Abraham, 2011

and Amin et al., 1996). It was reported to regulate steroidogenesis and improve semen quality in

infertile men (Shukla et al., 2009). Hexane extract of leaves of Moringa oleifera Lam. was used

by Cajuday and Pocsidio (2010) to investigate the fertility enhancing effect on male. In this

study, mice were fed with different doses of M. oleifera Lam. with oral gavage for 21 days. The

study showed that the weights of testis (at medium and high doses); epididymis (at all doses);

seminal vesicle (at the high dose) increased and also increased seminiferous tubule diameter (at

all doses); increased thickness of epididymal wall (at medium and high doses); higher score for

lumen formation (at the high dose) and epididymal maturity (at all doses) in the treated mice

when compared to the control. Serum level of LH and FSH were estimated which revealed no

significant difference with control (Cajuday and Pocsidio, 2010). In another study, sexual

function improving effect of Myristica fragrans Houtt. was reported in male rats. In this study,

rats were fed with different doses of 50% methanol extract of M. fragrans Houtt. for seven days

to study the effect of the plant on mating behavior, mounting frequency, penile reflexes and other

sexual performances. Results of the investigation showed significant increase in mounting

frequency, intromission frequency & intromission latency and caused significant reduction in the

mounting latency and post ejaculatory interval for high dose of the extract. It was also

documented to increase significantly mounting frequency with penile anaesthetisation as well as

erections, quick flips, long flips and the aggregate of penile reflexes with penile stimulation

(Tajuddin et al., 2005). Aphrodisiac and fertility enhancing effect of aerial parts Cynodon

dactylon was evaluated on male albino mice by Chidrawar et al., 2011. In this study,

experimental rats were subjected to stress induced sexual dysfunction and then treatment was

done with methanolic extract of C. dactylon. It was shown in the results that the extract had

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promising effect in overcoming stress-induced sexual dysfunction, sexual performance, fructose

content, sperm concentration and increased accessory sexual organs and body weight. Thus they

concluded that active constituents of C. dactylon present in methanolic extract have a potent

aphrodisiac and male fertility activity. In another study, role of alcoholic extract of Nigella sativa

seeds on fertility potential, pituitary-testicular axis hormones and testosterone in male rats was

investigated. In this study, male rats were fed with alcoholic seed extract for 60 days and fertility

parameters such as body and reproductive organs weight, sperm motility, viability and count,

epididymal sperm reserve, daily sperm production, blood testosterone concentration,

gonadotropins levels and fertility index were measured. Result of the study showed increased

fertility parameters along with the serum level of hormones indicating the fertility enhancing

property of the seeds of Nigella sativa (Parandin et al., 2012). Abelmoschus manihot L. seeds

were administered to male rats for 7 days to study the aphrodisiac activity by Rewatkar et al.,

2010. They have reported aphrodisiac property of the plant on the basis of increased sperm count

and penile erection index and on the basis of improved sexual behavior by increased mount and

intromission frequency.

An herbal combination of Withania somnifera, Tribulus terrestris, Mucuna pruriens and

Argyreia speciosa was reported to enhance reproductive capability in both sexes. Oral

administration of the herbal drug to both sexes of mice resulted in increase in reproduction rate

up to two generations i.e. F0 and F1 (Riaz et al., 2010).

Reproductive effects of Ficus asperifolia was studied in female rats by Watcho et al., 2009. In

this investigation, they used two types of extracts of the fruits of the plant, aqueous and

methanol. Reproductive effects were investigated on implantation study, fertility study and

uterotropic study. The outcome of the study revealed significant increase in the implantation sites

and litter size of animals receiving 100mg/kg of the aqueous extract of F. asperifolia, but there

was no significant difference in other treatment groups. In the estrogenic assay, it was shown that

normal immature rats were sensitive to the treatment with F. asperifolia than the ovariectomized

ones. Their results gave added scientific support to the popular use of F. asperifolia in the

treatment of some cases of women’s sterility/infertility related problems (Watcho et al., 2009).

Another species of Ficus was evaluated for fertility inducing effect on female rats. Seed, bark

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and leaves of the plant Ficus platyphylla Delile is traditionally used to promote fertility in the

Northern part of Nigeria. Aqueous extracts of stem bark, leaves and seeds of the plant were fed

to the experimental rats in three different doses for 20 days. The study revealed that there was an

increase in the number of litters in the experimental groups compared to the control. Thus it was

established that the plant is having fertility inducing effect on female (Chinenye et al., 2011).

Another plant Lepidium meyenii was found to increase litter size in mice (Ruiz-Luna et al.,

2005). Fertility inducing effect of aerial parts of Coccinia cordifolia L. in female rats was

evaluated by inducing infertility in the experimental animals. Infertility was achieved by three

ways, endometriosis induced infertility, testosterone induced infertility and hyperprolactinemia

induced infertility. In this investigation all the infertile animals were treated with aqueous extract

of aerial parts of the plant. Results showed the numbers of uterine implants were almost 10 times

more in the extract treated groups as compared to control in hyperprolactinemia induced

infertility model. The extract used in the study failed to induce fertility in rats with endometriosis

and there was no significant result in the testosterone induced infertility group. However, it was

reported that the extract produced a significant increase in serum estrogen levels and number of

corpus luteum in healthy female rats. Thus the aqueous extract of Coccinia cordifolia L. induces

fertility in hyperprolactinemia induced infertility model in female rats (Jha et al., 2010).

Analysis of fertility enhancing effects is studied not only in mammals but there are reports of

investigations of fertility enhancing effects of some plants in other animals also. Ethanol extracts

of Garcinia kola was tested as fertility enhancer in female catfish Clarias gariepinus brood

stock. There were significant increase in the weight, fecundity and egg size of the extract treated

fishes (Dada and Ajilore, 2009). The investigators thus concluded the study that the ethanol

extract of G. kola seeds possesses promising pro-fertility property which can be exploited in fish

seeds production. In another experiment dried Kigelia africana fruits meal was used to enhance

fertility in female Clarias gariepinus. It was observed that the fishes fed with Kigelia africana

fruits meal decreased % of deformity in hatchlings, increased % of survival and increase in the

egg size (Dada et al., 2010). Kigelia africana was also evaluated for male fish also. The male

brood fish fed with K. africana fruits significantly increased sperm counts, increased % in sperm

motility, increased fertilization ability, lower milt volume and increased sperm motility duration

(Adeparusi et al., 2010). Thus there are enough reports on plants having fertility enhancing

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properties. Table-2.1: represents some of the fertility enhancing plants scientifically evaluated as

well as traditionally used in different parts of the world.

Table-2.1: List of fertility enhancing plants used worldwide:

Sl

no.

Name of the plant. Family. Parts

used.

Purpose of use/

Disorders.

References.

1. A. kerstingii, Carpolobia

alba G. Don. and C.

lutea G. Don.

Leguminaceae Polygalaceae Polygalaceae

Roots.

Aphrodisiac.

Muanya and Odukoya, 2008.

2. Abelmoschus manihot. Malvaceae. Seeds. Male Aphrodisiac Rewatkar et al., 2010.

3.

Abroma augusta L.F.

Malvaceae Leaves, Stems, Bark of roots

Menstrual disorders, Uterine diseases, Leucorrhoea

Hossan et al., 2010.

4. Acanthus ilicifolius L. Acanthaceae Roots Leucorrhoea Hossan et al., 2010.

5. Acanthus montanus Acnthaceae Leaf. Enhances female fertility

Focho et al., 2009.

6.

Achyranthes aspera L. Amaranthaceae.

Leaf.

Induce labour Leucorrhoea, menstrual problems

Sharma and Sharma, 2010. Hossan et al., 2010.

7. Aerva javanica (Burmf.) Schultes

Amaranthaceae Whole plant.

Aphrodisiac Elkhalifa et al., 2006.

8. Aframomum danielli K. Schum

Zingeberaceae Roots Gonorrhea, Male infertility.

Focho et al., 2009.

9. Aframomum melegueta Zingiberaceae Seed Male and Female infertility.

Okoli et al., 2007; Focho et al., 2009.

10. Aloe barbadensis Linn. Liliaceae Leaves Female infertility. Focho et al., 2009.

11. Aloe buettneri. A Berger & Eng

Liliaceae Leaves Female infertility. Focho et al., 2009.

12. Amaranthus spinosus L. Amaranthaceae. Leaf. Increase Lactation.

Sharma and Sharma, 2010.

13. Ambrosia maritime Linn. Asteraceae Leaf.

Female infertility. Focho et al., 2009.

14. Anthocleista djalonensis A. Chev.

Leguminaceae Roots. Aphrodisiac. Muanya and Odukoya, 2008.

15.

Argemine Mexicana Linn.

Papaverraceae

Seeds and plant latex.

Impotence

Ghosh, 2008.

16.

Argyreia nervosa

Convolvulaceae

Leaf, roots & flowers

Male Aphrodisiac.

Subramoniam et al., 2007.

17. Basella alba Linn Basellaceae Whole plant


18. Bidens pilosa Linn Asteraceae Whole plant

Male and Female infertility.

Focho et al., 2009.

19. Boerhaavia diffusa Nyctaginaceae. Leaf Increases fertility. Okoli et al., 2007.

20. Cannabis sativa L. Cannabaceae Leaf, seed

Sex stimulant. Nawaz et al., 2009.

21. Cassia sieberiana DC Leguminaceae Roots. Aphrodisiac. Muanya and Odukoya,

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2008.

22. Centella asiatica (L.) Urban

Apiaceae Whole plant

Leucorrhoea. Female infertility.

Hossan et al., 2010. Focho et al., 2009.

23. Chasmanthera

dependens Hochst. Menispermaceae Roots. Aphrodisiac. Muanya and Odukoya,

2008.

24. Cissus populnea Guill and Perr.

Vitaceae Roots. Aphrodisiac. Muanya and Odukoya, 2008.

25 Cissus quadrangularis Linn

Vitaceae Whole plant


Focho et al., 2009.

26. Citrus sinensis (L). Obs Rutaceae Roots Impotence. Focho et al., 2009.

27. Cleome viscose Capparaceae Leaf Female infertility. Okoli et al., 2007.

28. Clitoria ternetea L. Fabaceae Roots Impotency Infertility. Aphrodisiac.

Das et al., 2008: Ghosh, 2008: Fantz, 1991.

29. Cnestis ferruginea DC Connaraceae Roots. Aphrodisiac. Muanya and Odukoya, 2008.

30. Coccinia cordifolia L. Cucurbitaceae Aerial parts Roots

Female infertility. Leucorrhoea, menstruation problems

Jha et al., 2010. Hossan et al., 2010.

31. Cola nitida Schott and Bandl

Stercliaceae Fruit Male and Female Infertility

Focho et al., 2009.

32. Costus speciosus (Koen.) Sm.

Costaceae Roots Leucorrhoea Hossan et al., 2010.

33. Crassocephalum biafrae. S. Moore.

Asteraceae Whole plant

Male and Female Infertility

Focho et al., 2009.

34. Crateva adansoni Capparidaceae Root. Male sexual stimulant.

Okoli et al., 2007.

35. Cyathea manni Hook Cyatheaceae Bark Oligospermia Impotence

Focho et al., 2009.

36. Cynodon dactylon Poaceae Aerial part

Male infertility. Chidrawar et al., 2011.

37. Cynodon dactylon (L.) Pers. Var. glabratus (Steud.) Chiov.

Poaceae Leaf, stem

Menstrual problems.

Nawaz et al., 2009.

38. Dehaasia kurzii King ex Hook. f.

Lauraceae Leaves, roots

Leucorrhoea Hossan et al., 2010.

39. Discorea cayenensis Lam.

Discoreaceae Roots. Aphrodisiac. Muanya and Odukoya, 2008.

40. Dychoriste perrotteti (Nees) O. Ktze

Acanthaceae Leaves Female infertility. Focho et al., 2009.

41. Eclipta alba (L.) Hassk Asteraceae Whole plant


42. Emblica officinalis

Gaertn. Euphorbiaceae Leaves,

fruit Leucorrhoea Hossan et al., 2010.

43. Emilia coccinea (Sims) G. Don

Asteraceae Whole plant


Focho et al., 2009.

44. Entada africana Guill &Perr

Mimosaceae Bark Female infertility. Focho et al., 2009.

45. Eremomastrax speciosa (Hochst) Cufod.

Acanthaceae Leaves Male and Female infertility.

Focho et al., 2009.

46. Eugenia jambolana Lamk.

Myrtaceae. Seeds Leucorrhoea. Sharma and Sharma, 2010.

47. Euphorbia hirta Linn Euporbiaceae Leaf twigs

Female infertility Focho et al., 2009.

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48. Eurycoma longifolia Jack.

Simaroubaceae. Roots. Male sexual enhancer.

Ang et al., 2000.

49.

Ferula assa-foetida L. Apiaceae

Seeds and roots

Enhances male fertility, Impotence and Aphrodisiac.

Kassis et al., 2009.

50. Ficus asperifolia. Moraceae. Fruits. Female infertility Watcho et al., 2009.

51. Ficus exasperata Vahl Moraceae Buds, bark


52.

Ficus platyphylla

Moraceae

stem bark, leaves and seeds

Enhance female fertility.

Chinenye et al, 2011.

53. Foeniculum vulgare Mill.

Apiaceae. Fruits Increases Folliculogenesis Increases Libido

Khazaei et al., 2011; Onyiapat et al., 2011.

54. Garcinia kola. Guttiferae Seeds. Fertility enhancer in female catfish.

Dada and Ajilore, 2009.

55. Ginkgo biloba Ginkgoaceae Leaves Aphrodisiac; Male fertility enhancer.

Huat Chye, 2006; Yeh et al., 2008.

56. Glycosmis pentaphylla (Retz.) DC.

Rutaceae Whole plant


57. Herbal Combination (Withania somnifera,

Tribulus terrestris,

Mucuna pruriens and Argyreia speciosa).

Solanaceae, Zygophyllaceae, Fabaceae and Convolvulaceae.

Fertility enhancer in male and female mice.

Riaz et al., 2010.

58. Hibiscus asper Hook. F Malvaceae Leaves Male and Female infertility.

Focho et al., 2009.

59.

Hibiscus rosachinensis L.

Malvaceae

Flowers

Increases sperm number. Menstrual Pain Relief.

Sharma and Sharma, 2010. Acharyya and Sharma, 2004.

60. Holarrhena pubescens Buch.-Ham. Wall. ex G. Don

Apocynaceae

Bark

Leucorrhoea


61. Hygrophila spinosa T. Anders.

Acanthaceae Whole plant


62. Ipomoea paniculata Burm.f.

Convolvulaceae

Whole plant

Leucorrhoea, menstrual problems.


63. Kaempferia parviflora Zingiberaceae Rhizomes

Male sexual enhancer.

Chaturapanich et al., 2008.

64.

Kigelia Africana

Bignoniaceae

Fruits.

Fertility enhancer in Male and Female Catfish. Infertility.

Dada et al., 2010. Focho et al., 2009.

65. Lawsinia alba Lamb. Lythraceae. Leaves Dysmenorrhoea. Sharma and Sharma, 2010

66. Lecaniodiscus

cupanioides Planch Sapindaceae Roots. Aphrodisiac. Muanya and Odukoya,

2008.

67. Leea guineensis G. Don. Leeaceae Bark Male and Female infertility.

Focho et al., 2009.

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68. Leonotis nepetifolia (Linn) Ait.f.

Lamiaceae Whole plant

Female infertility Focho et al., 2009.

69.

Lepidium meyenii

Brassicaceae

Roots.

Male and female fertility enhancer.

Gonzales et al., 2001a, 2001b and 2003. Cicero et al., 2001 & 2002. Ruiz-Luna et

al., 2005.

70. Litsea liyuyingi H. Liu Lauraceae Leaves, bark


71. Lophira lanceolata. Ochnaceae Stem Bark

Male fertility enhancer

Etuk and Muhammad, 2009.

72. Malachra capitata Linn. Malvaceae Fruits Infertility Ghosh, 2008.

73. Markhamia tomentosa (Ben.) K. Sch. Ex. Engl

Bignoniaceae Bark Male and Female infertility.

Focho et al., 2009.

74. Microdermis keayana J. Leonard.

Pandaceae Roots. Aphrodisiac. Muanya and Odukoya, 2008.

75. Mimosa pudica Linn. Minosaceae Roots Infertility Ghosh, 2008.

76. Momordica charantia Cucurbitaceae Leaf, fruits.

Fertility enhancer. Okoli et al., 2007.

77. Mondia whitei Skeels Periplocaceae Roots Male infertility Focho et al., 2009.

78. Moringa oleifera Lam. Moringaceae Leaves Male Fertility Enhancer

Cajuday and Pocsidio, 2010.

79.

Mucuna pruriens L. Fabaceae

Seeds

Male infertility. Shukla et al., 2009; Abraham, 2011; Amin et al., 1996.

80. Myristica fragrans Houtt.

Myristicaceae Kernel Male Aphrodisiac Tajuddin et al., 2005.

81.

Nigella sativa L. Ranunculaceae Seeds Enhances male fertility.

Parandin et al., 2012; Al-Saaidi et al., 2009.

82. Ocimum sanctum L. Lamiaceae Seed Gonorrhea Nawaz et al., 2009.

83.

Odontotermes

formosanus (Termite)

Termitidae

Whole body.

Male and Female fertility Enhancer. Increases Lactation.

Solavan et al., 2006 and 2004.

84. Oroxylum indicum (L.) Kurz

Bignoniaceae Bark, fruit

Leucorrhoea, urinary problems


85. Panax ginseng Araliaceae - Aphrodisiac Kim et al. 1976. Nocerino et al., 2000.

86. Plectranthus glandulosus Hook. F

Lamiaceae Leaves Female infertility. Dysmenorrhoea

Focho et al., 2009.

87. Pseudospondias

microcarpa Anacadiaceae Bark Female infertility Focho et al., 2009.

88. Psidium guajava Linn Myrtaceae Roots Impotence Focho et al., 2009.

89. Pteridium aquilinum (Linn). Kohn.

Dennstaedteaceae

Buds Male infertility Focho et al., 2009.

90. Raphia hookeri Arecaceae Sap Male and Female infertility

Focho et al., 2009.

91. Ruellia praetermissa Acanthaceae Leaves Maintenance of pregnancy.

Salah and Wagner, 2009.

92. Senna alata (Linn). Roxb

Caesalpinaceae Leaves Female infertility Focho et al., 2009.

93. Sida cordifolia L. Malvaceae Leaf, bark of roots

Increases sperm number.

Nawaz et al., 2009.

94. Sida veronicifolia Lam. Malvaceae Whole Male infertility. Focho et al., 2009.

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plant Dysmenorrhoea.

95. Solanum aculeastrum Solanaceae Fruit Female infertility. Focho et al., 2009.

96. Solanum nigram L. Solanaceae Seeds Aphrodisiac Elkhalifa et al., 2006.

97. Spathodea campanulata P. Beauv

Bignoniaceae Bark Male and Female infertility

Focho et al., 2009.

98. Sphenocetrum jolyanum Menispermaceae Roots Impotence Okoli et al., 2007.

99. Spilanthes acmella Asteraceae Leaves, flowers


100 Spilanthes filicaulis (Schum & Thonn) C.D. Adams

Asteraceae

Leaves

Impotence.

Focho et al., 2009.

101. Sterculia tragacantha Lindl

Sterculiaceae Bark Male and Female infertility

Focho et al., 2009.

102 Tabernaemontana C.F ventricosa Hochst.ex. A.D.C.

Apocynaceae Bark


Focho et al., 2009.

103 Telferia occidentalis Curcubitaceae Seeds Increases sperm number.

Okoli et al., 2007.

104 Tribulus terrestris L. Zygopyllaceae. Leaves. Aphrodisiac. Sharma & Sharma, 2010;Huat Chye, 2006

105 Triumfetta rhomoboidea Malvaceae. Leaf Induce fertility. Okoli et al., 2007.

106 Vernonia ambigua Kotschy & Peyr.

Asteraceae Leaves Male and Female infertility.

Focho et al., 2009.

107 Vernonia amygdalina Del.

Asteraceae Roots Impotence. Focho et al., 2009.

108 Vernonia cinerea (L.) Less.

Asteraceae Leaves Leucorrhoea Das et al., 2008.

109 Vernonia guineensis Benth

Asteraceae Roots Male and Female infertility.

Focho et al., 2009.

110 Withania somnifera Dunal.

Solanaceae. Leaf. Enhances female fertility.

Saritha et al., 2011.

111 Xanthosoma

sagittaefolium Schott. Araceae Tubers. Female infertility. Focho et al., 2009.

2.2: PLANT COMPOUNDS RESPONSIBLE FOR ENHANCING FERTILITY:

Plants can be the remedies for infertility in men and women (aphrodisiac), for menstruation

problems including irregular menstruation, for promotion of menstruation, for painful

menstruation, for excessive or prolonged uterine bleeding, for absence of menstruation and for

pregnancy related problems. All plants contain number of compounds such as nutrients, minerals

and food sources etc. which contribute to improvement of fertility. Good nutrition and a healthy

lifestyle can have a positive effect on fertility and childbearing. How nutritional factors might

influence fertility is largely an unexplored field. Very little is known about how dietary

composition influences fertility. Vitamins, minerals, food staffs and specific cofactors play a

major role in fertility function (Westphal et al., 2004). Vitamin B6 has been shown to improve

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conception rates while Vitamin B12 (Bennett, 2001), Vitamin E (Bayer, 1960) and multivitamins

(Czeizel et al., 1996) have been shown to improve female fertility. Human and animal data

suggest that low vitamin D status is associated with impaired fertility, endometriosis and

polycystic ovary syndrome. Evidence from observational studies shows higher rates of

preeclampsia, preterm birth, bacterial vaginosis and gestational diabetes in women with low

vitamin D levels (Grundmann and von Versen-Hoynck, 2011). Experiments documented that

vitamin-D deficient female rats had reduced fertility rates, decreased litter sizes and

compromised mating behavior (Halloran and DeLuca, 1980). Vitamin A has been reported to

have positive effect on both male and female reproduction (Thompson et al., 1964). It was also

reported to have effect on embryonic development (Clagett-Dame and DeLuca, 2002).

Minerals of plant origin have been reported to increase fertility. Minerals such as folic acid

(Dawson and Sawers, 1982), magnesium, selenium (Howard et al., 1994), iron (Rushton et al.,

1991), copper and zinc (Bedwal and Bahuguna, 1994) have been documented to improve female

fertility. It has been reported that an adequate intake of essential nutrients such as folic acid, in

the periconceptual period can lower the incidence of neural tube defects (Westphal et al., 2004).

Copper is needed by a variety of key systems in the body. Numerous enzymes necessary for

reproduction, immunity and growth need copper. In addition, copper is necessary for proper

metabolism of iron, maintenance of connective tissue, pigmentation of skin and hair, maturation

of hoof tissue, and many other functions. It is widely known that copper deficiency results in

reduced reproductive efficiency and performance. It is theorized that low copper levels alter

enzyme systems involved in reproduction. Typical copper deficiency symptoms include

decreased conception rates, decreased libido and semen quality. Zinc has been reported to use in

the treatment of male infertility for increasing sperm number and motility. It was also stated that

zinc is necessary during pregnancy and lactation (Bhowmik et al., 2010). Manganese stimulates

secretion of LH necessary for ovulation. It was reported that manganese chloride when

administered acutely into the third ventricle of the brain acts dose-dependently to stimulate LH

release in prepubertal female rats (Pine et al., 2005). Supplementation with copper, zinc and

manganese improved pregnancy rate in grazing cattle (Ahola et al., 2004). Magnesium

supplementation significantly increased androgenic enzymes along with enhanced functioning of

testis (Chandra et al., 2013). It has been reported that magnesium plays a role in ovule

35

maturation, sperm viability and fecundation in Invertebrates and Fishes. In Birds and Mammals,

it appears to be necessary for fecundation. In rat, the pregnancy cannot be normal, unless the

food contains an adequate supply of magnesium. Severe or mild deficiencies affect the site of

fetal implantation and, if they are prolonged, lead to abortion in the first instance and

pathological disorders in the latter (Stolkowski, 1977).

The three major food staffs protein, carbohydrate and lipid influence fertility directly or

indirectly. All the three components of food help to enhance fertility or to overcome some of the

fertility disorders. Amino acid like L-arginine helps improving circulation to the reproductive

organs that may enhance oocyte development and implantation of the embryo. Insulin sensitivity,

which can be affected by diet, is an important determinant of ovulatory function and fertility

(Azziz et al., 2001). The amount and sources of protein in diet have been found to influence

insulin sensitivity (Gannon et al., 2003). Increasing vegetable (soy) protein intake has been

found to increase ovulation rates in pigs (Mejia-Guadarrama et al., 2002). In another study it

was stated that, replacing animal sources of protein with vegetable sources of protein may reduce

ovulatory infertility risk (Chavarro et al., 2008). Like proteins, carbohydrates are also necessary

for successful fertility as it gives required energy. A cascade of cell–cell interactions is required

to achieve fertilization in mammals. Carbohydrates are required mainly for the biochemical

events in fertilization. Initial sperm–egg binding in mammals involves recognition of

glycosylated proteins of egg zonae by glycosylated proteins on sperm surfaces (Benoff, 1997).

Lipids include cholesterol, phospholipids and triacylglycerols. These and their derivatives

provide energy and are also essential components in a variety of endocrine and cell signaling

pathways (Mattos et al. 2000; Wathes et al. 2007b). Research shows if there is a reproductive

benefit to supplementing fat it is probably prepartum, targeting those animals in most need, such

as the young growing females that are carrying a child for the first time (Haag, 2008). Adequate

intake of monounsaturated fatty acids, derived mainly from vegetable fats, as well as avoidance

of trans-isomers of unsaturated fatty acids which are present in industrially produced cakes and

sweets, crisps, fast-foods, powdered soups and hard margarines, may be effective in the

prevention of infertility in females (Szostak-Węgierek, 2011).

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A number of plant constituents are chemically or structurally similar to natural hormones and can

be used as substitute in case of hormone deficiency. Different phytochemical groups such as

flavonoids, saponins, alkaloids and phenols have been documented for their effects on

reproduction of animals. Plant components often have an indirect action on the secretion of

certain hormones by stimulating or inhibiting the hypothalamus and pituitary gland and in this

way controlling the functions of most other glands. Alkaloids are a group of naturally occurring

chemical compounds that contain mostly basic nitrogen atoms and produced by a large variety of

organisms, including bacteria, fungi, plants, and animals. Role of alkaloids have been

documented for both fertility enhancing and anti- fertility properties. Many alkylating agents

have been found to produce infertility, both in man and in animals (Miller, 1971; Fairley et al.,

1972). Quinolizidine alkaloids were isolated from the roots of Cnestis ferruginea and found

significant reduction in sperm counts, motility, viability, morphology and plasma levels of

testosterone, luteinizing hormone and follicle stimulating hormone, indicating the anti-fertility

activity (Olayemi and Raji, 2011). Anti-fertility effects of alkylating agents and vinca alkaloids

in male rats were reported by Cooke et al., 1978. Some alkaloids have been documented to have

positive effects on fertility. Dr. Gloria Chacon de Popivici, a biologist trained at the University of

San Marcos, in Lima, Peru isolated four alkaloids from the maca root and carried out animal

studies with male and female rats. They were given either powdered maca root or alkaloids

isolated from the roots. In comparison with the animal control groups, those receiving either root

powder or alkaloids showed multiple egg follicle maturation in females and, in males,

significantly higher sperm production and motility rates than control groups. Meaning, the

alkaloids in the maca root (not its plant hormones) produced fertility effects on the ovaries and

testes of the rats. The alkaloid yohimbine is found in the bark of a tree Pausinystalia johimbe (K.

Schum) Beille that grows in Central Africa and which has been used for centuries as an

aphrodisiac. Studies revealed that yohimnine , in combination with drugs that facilitate the action

of nitric oxide, is effective in the treatment of male erectile dysfunction and when combined with

L-arginine glutamate, increase sexual arousal in women. This alkaloid slightly raises serum

testosterone level in male (Maggi et al., 2000). Plants used to treat dysmenorrhoea, such as

Alchornea cordifolia Mull and some members of the Solanaceae family contain atropine,

papavarine and yohimbine. Some forms of dysmenorrhoea are treated with plants such as parsley

which contain essential oils that produce contractions of uterus and are abortive in high doses

37

(Oliver-Bever, 1986). Thus it can be seen that alkaloids have both positive and negative effect on

reproduction. Polyphenols are naturally occurring compounds found largely in the fruits,

vegetables, cereals and beverages. Polyphenols are classified on the basis of the number of

phenol rings that they contain and of the structural elements that bind these rings to one another

(Fig: 2.1). They are broadly divided in four classes; Phenolic acids, flavonoids, stilbenes and

lignans (Pandey and Rizvi, 2009). Favonoids comprise the most studied group of polyphenols.

This group has a common basic structure consisting of two aromatic rings bound together by

three carbon atoms that form an oxygenated heterocycle (Fig: 2.2). Flavonoids have diverse

effects on both male and female fertility. Coumestans and isoflavones, which differ in chemical

structure from estrogenic hormones, have been shown to have activity similar to these hormones

(Ducan et al., 2003). It is because of this activity that they may have beneficial as well as adverse

effects. Many plants have been reported to contain fertility enhancing effects and all the plants

have flavonoid in their phytochemical composition. Such plants which have been reported to

enhance fertility in male include Cissus populnea and Panax ginseng (Oremosu et al., 2013);

Argemine mexicana Linn. (Ghosh, 2008); Crateva adansoni (Okoli et al., 2007); Ginkgo biloba

(Huat Chye, 2006); Lepidium meyenii (Gonzales et al., 2001a, 2001b and 2003) etc. Plants which

have been reported to enhance female fertility include Lepidium meyenii (Ruiz-Luna et al.,

2005); Vernonia guineensis Benth (Focho et al., 2009); Withania somnifera Dunal. (Saritha et

al., 2011); Ficus asperifolia (Watcho et al., 2009); Discorea cayenensis Lam. (Muanya and

Odukoya, 2008) etc. From this observation it is evident that flavonoids have positive role in

fertility. Saponins are chemical structures consisting of triterpenoidal or steroidal aglycones with

various carbohydrate moieties that are found in many plants. Saponins have been reported to

improve fertility in both, male and female. Korean red ginseng total saponins have been reported

to improve fertility in females with PCOS (Pak et al., 2005). Panax ginseng C.A. Meyer

saponins have also been reported to enhance male fertility (Salvati et al., 1996). All the

phytochemicals mentioned above have fertility enhancing property and some of them have anti-

fertility property also. Plants which have been documented by different scientists contain all or

some of the phytochemicals mentioned here in different concentrations. Therefore, it is necessary

to estimate these phytochemicals before undertaking any animal experiments.

38

Fig. 2.1: Chemical structures of the different classes of polyphenols (from Pandey and Rizvi, 2009).

Fig. 2.2: Chemical structures of the flavonoid group (from Pandey and Rizvi, 2009).

39

2.3: ABOUT THE PLANT USED IN THE PRESENT STUDY:

The present study was designed to evaluate fertility enhancing effect of a selected plant

documented in the survey (Plate-1.2-A). The plant belongs to the genus Clitoria of family

Fabaceae (Plate-1.2-B), which comprises 60 species distributed mostly within the tropical belt

with a few species found in temperate areas (Gupta et al., 2010). Clitoria ternatea Linn is

commonly known as Aparajita (Assamese), Koyala (Hindi), Sangu pushpam (Malayalam),

Butterfly pea (English) etc. Aparajita is one of the herbs mentioned in all ancient scriptures of

Ayurveda. It has several synonyms in Ayurvedic scriptures like Gokarnika, Ardrakarni,

Vishnukranta, Girikarnika, Supuspi, Mohanasini, Sveta etc.

C. ternatea L. is a vigorous, strongly persistent, herbaceous perennial legume having twining

fine stems, 0.5-3 m long. The leaves are pinnate, with 5-7 elliptic to lanceolate leaflets, 3-5 cm

long and shortly pubescent underneath. Flowers are axillary, single or paired, colour ranges from

white, mauve, light blue to dark blue (Plate-2.1-C) with very short pedicellate and 4-5 cm long.

Pods are flat, linear, beaked, 6-12 cm long, 0.7-1.2 mm wide and slightly pubescent with up to

10 seeds. The seeds are olive, brown or black in colour, often mottled, 4.5-7 mm long and 3-4

mm wide (Hall, 1985). The root system of Clitoria ternatea L. consists of a fairly stout taproot

with few branches and many slender lateral roots. Multicellular trichomes, with two basal cells

smaller than the terminal cells are present. In Transverse section leaf shows a dorso-ventral

structure. All along the veins prismatic crystals of calcium oxalate are present. The vein–islet

number is 7.5 and palisade ratio is 6.0. The pods are (5-10 cm) long, flat and 6-11 seeded

(Mukherjee et al., 2008; Karandikar and Satakopan, 1959). Cortex is composed of 10- 12 layers

of thin-walled almost polygonal or tangenially elongated cells, packed with mostly compound

starch grains (Mukherjee et al., 2008; Shah and Bole, 1961). All the ray cells are fully packed

with starch grains and few contain calcium oxalate crystals (Mukherjee et al., 2008; Kalamani

and Michael, 2001 & 2003). Almost all parts of this plant are reported to have medicinal

properties. Flowers of this plant has been using in a number of Hindu religious purposes since

the ancient times.

A. Whole Plant of Clitoria ternatea

C. Different Flower Varieties of

D. Plant part used in the present study.

Plate-2.1: Whole plant (A), Systematic position (B),

part used (D)of the plant C. ternatea

40

ternatea L. B. Systematic position of the plant.

C. Different Flower Varieties of C. ternatea L. available in Assam.

D. Plant part used in the present study.

(A), Systematic position (B), Different flower varieties (C)

C. ternatea L. selected in the study.

Root of the Plant

Root powder

B. Systematic position of the plant.

Different flower varieties (C) and plant

41

C. ternatea L. is distributed almost throughout India, wild or cultivated, Western - north India,

Egypt, Syria, Mesopotamia, Iraq, Persia, Arabia, Afghanistan etc. It can be grown as a forage

legume either alone or with perennial fodder grasses in Punjab, Rajasthan, Uttar Pradesh,

Gujarat, Maharashtra, Madhya-Pradesh, Andhra-Pradesh and Karnataka. The plant is also

suitable as a green manure and cover crop (Gupta et al., 2010).

2.3.1: Traditional uses of C. ternatea L.:

Clitoria ternatea L is a very well-known Ayurvedic medicine used for different ailments, which

has been investigated scientifically in considerable detail. The disease preventive and health

promoting approach of Ayurveda, takes into consideration of the whole body, mind and spirit

while dealing with the maintenance of health, promotion of health and treating ailments is

holistic way and finds increasing acceptability in many regions of the world (Mukherjee et al.,

2007-b). From ancient times “Shankhpushpi” is known as reputed drug of Ayurveda and

reported as a brain tonic, nerve tonic and laxative. It is considered as a “Medhya-Rasayana” in

Ayurvedic texts that comprises with herbs like Convolvulus pluricaulis (Convolvulaceae),

Evolvulus alsinoides (Convolvulaceae), Clitoria ternatea (Fabaceae) and Conscora decusata

(Gentianaceae). It is an Ayurvedic drug used for its action on the central nervous system,

especially for boosting memory and improving intellect (Sethiya et al., 2009; Kumar et al.,

2008). The flowers of the plant C. ternatea L. resemble a conch shell; therefore it is commonly

called “Shankpushpi” in the Sanskrit language where it is reported to be a good “Medhya” (brain

tonic) drug and, therefore, used in the treatment of “Masasika Roga”, which means mental illness

(Daisy and Rajathi, 2009). Traditionally the leaves and roots are used in the treatment of a

number of ailments including body aches, especially infections, urino-genital disorders, and as

an anthelmintic and antidote to animal stings. The root is used in the treatment of various

diseases, like indigestion, constipation, fever, arthritis and eye ailments (Mukherjee et al., 2008).

It is also employed in cases of ascetics, enlargement of the abdominal viscera, sore throat and

skin diseases (Anonymous, 1995). They are also demulcent and given in chronic bronchitis.

Though they are purgative, they cause griping and tenderness, and hence are not recommended.

They are, however, administered with honey and ghee as a general tonic to children for

improving mental faculties, muscular strength and complexion tonics and in epilepsy and

insanity (Anonymous, 1976). The root-juice of the white-flowered variety is blown up the

42

nostrils as a remedy for hemicrania. The decoction or powder of root is given in rheumatism, and

ear-diseases. Powdered seeds are mixed with ginger and given as laxative, the action, however, is

accompanied by griping in lower abdomen. The seeds are considered for colic, dropsy and

enlargement of abdominal viscera; they are also used in swollen joints (Morris, 1999 and

Anonymous, 2001). The root, stem and flower are recommended for the treatment of snakebite

and scorpion sting in India (Kirtikar and Basu, 1935). In Cuba decoction of roots alone or roots

and flowers are considered emmenagogue. This mixture is made by placing a handful of cleaned

and macerated roots in a bottle of water. A glass taken in the evening is said to promote

menstruation and induce uterine contractions. A stronger dose of the same liquid is used as a

vaginal douche (Mukherjee et al., 2008). The root juice is given in cold milk to remove the

phlegm in chronic bronchitis. It causes nausea and vomiting. The roots and seeds prescribed

internally in cough, disease of liver, spleen and rheumatic affections (Patil and Patil, 2011). The

roots are used to treat impotency & infertility and said to have aphrodisiac property in both male

and female individuals (Das et al., 2008; Ghosh, 2008; Fantz, 1991).

2.3.2: Pharmacological Activities of C. ternatea L.:

Scientific evaluation of the plant C. ternatea L. on its medicinal properties have confirmed that it

exhibits a broad range of biological effects, some of which are very interesting for promising

future development. Medicinal properties of different parts of C. ternatea L. evaluated so far are

as follows:

A. Effect of C. ternatea L. on CNS:

Memory enhancement activity: Different parts of the plant C. ternatea L. have been tested for

memory enhancement activity by different workers. The oral treatment of C. ternatea L. roots

extract was reported to increase significantly memory in rats (Rai et al., 2001). The alcoholic

extracts of aerial parts and root of C. ternatea L. was reported to attenuate electroshock induced

amnesia. The acetylcholine (AcH) content of the whole brain and acetyl cholinesterase activity at

different regions of the rat brain viz. cerebral cortex, mid-brain, medulla oblongata and

cerebellum was evaluated (Taranalli and Cheeramkuzhy, 2000). It was suggested that an increase

in AcH in rat hippocampus may be the neurochemical basis for improved learning and memory

(Rai et al., 2002 & Mukherjee et al., 2007). Rai et al., 2000 have also shown that the aqueous

43

root extract of C. ternatea L. enhances memory in rats. In another reported study the effect of

aqueous root extract on the dendritic cytoarchitecture of neurons of the amygdale was studied.

This improved dendritic arborisation of amygdaloidal neurons correlates with the increase

passive avoidance learning and memory in the C. ternatea L. treated rats (Rai et al., 2005).

Nootropic activity: The methanolic extract of aerial parts of C. ternatea L. was studied for

nootropic activity. The nootropic activity of C. ternatea L. was screened by using elevated plus

maze and object recognition test. The transfer latency was expressed as inflexion ratio. C.

ternatea L. increased the inflexion ratio significantly on ninth day only. The nootropic drugs

facilitate intellectual performance, learning and memory. C. ternatea L. and piracetam treated

mice required significantly less time to explore the familiar object as compared with the new

object. Both C. ternatea L. and piracetam (standard drug) significantly reduced the

discrimination index (Itoh et al., 1990).

Anticonvulsant activity study: The anticonvulsant activity of C. ternatea L. was assessed using

maximum electroshock induced seizures in mice. The C. ternatea L. and diazepam significantly

delayed the onset of convulsions. The animals treated with C. ternatea L. and diazepam

exhibited reduction in the duration of tonic hind limb extension. The extensor phase was reduced

after treatment of C. ternatea L. which showed anticonvulsant property (Swinyard and

Woodhead, 1982).

Anxiolytic activity: Anxiolytic activity was assessed by using elevated plus maze and the

light/dark exploration test. The oral administration of C. ternatea L. dose dependently increased

the time spent in the open arm (Lister, 1987).

Antidepressant activity: Antidepressant activity was studied by tail suspension test. Mice were

divided in groups of five each. They were suspended by tying a thread to their tail from a height

of 50cm above the table top. Duration of immobility was recorded for 6 min. Oral administration

of C. ternatea L. significantly reduced the duration of immobility (Steru et al., 1985).

Anti-stress activity or anti-histaminic activity: Anti-stress activity of C. ternatea L. was

evaluated by using cold restraint stress-induced ulcers and lithium-induced head twitches

methods by Alphine and Word (1969). In their study, rats were divided into groups of five each

and fasted for 18 h. Rats were treated with vehicle, different doses of C. ternatea L. 60 min

before and diazepam, 30 min before the test. Each rat subjected to CRS and stress was observed

through gastric ulcer index. The treatment with C. ternatea L. and diazepam significantly

44

reduced the ulcer index (Alphine and Word, 1969). Antihistaminic activity of the root of the

plant was also evaluated using clonidine and haloperidol induced catalepsy in mice. Finding of

investigation showed that chlorpheniramine maleate and ethanol extract of C. ternatea L. root

inhibit clonidine induced catalepsy significantly when compare to control group, thus proved the

presence of antihistaminic activity in the roots of C. ternatea L. (Taur and Patil, 2011).

B. Antipyretic and Analgesic Activities: Ethanolic extract of C. ternatea L. was evaluated for

analgesic activity in mice with the acetic acid-induced writhing response and mechanical

stimulus by tail clip method (Parimaladevi et al., 2003). In another study, the methanol extract of

C. ternatea L. was evaluated for its anti-pyretic potential in albino rats and the anti-pyretic effect

of the extract was comparable to that of paracetamol a standard anti-pyretic agent (Parimaladevi

et al., 2004).

C. Anthelmintic Properties: There are number of studies which have been reported on

antihelmintic activity of C. ternatea L. Alcoholic extract of roots of C. ternatea L. at various

concentrations and its various fractions were tested for anthelmintic activity against Indian

earthworm pheretima postuma. In this study Piperazine citrate was used as a standard reference

and distilled water as control. The result of the study indicated that the crude alcoholic extract

and its ethyl acetate as well as methanol fractions significantly demonstrated paralysis and also

caused death of worms especially at higher concentration as compared to standard reference

piperazine citrate (Khadatkar et al., 2008). Inhibitory effect of C. ternatea L. leaves on free-

living nematodes was evaluated using aqueous and methanol extract (Das et al., 2006; Nahar et

al., 2010). In another study, flowers, leaves, stems and roots of C. ternatea L. were evaluated for

anthelmintic activity on adult Indian earthworms Pheretima posthuma. Methanol extract of root

was found to be most potent and required very less time to paralysis and death of worms as

compared to other extracts. The potency increases from flowers, leaves, stems to roots (Nirmal et

al., 2008). Anthelmintic property of the leaf was investigated with both aqueous and ethanol

extracts (Salhan et al., 2011).

D. Anti-microbial activities: The methanolic extracts of the leaves and root of C. ternatea L.

were tested for their antibacterial activity against different pathogenic drug resistant Gram-

45

positive and Gram-negative clinical isolates. Minimum inhibitory concentration was determined

by agar dilution technique followed by estimation of zone of inhibition against the selected

strains by disc diffusion technique and comparison was done with reference to the standard

antibiotic ciprofloxacin. The leaf was found to possess powerful antibacterial activity against

Escherichia coli and Vibrio cholerae known for causing dysentery and Staphylococcus aureus

causative agent of fever. The leaf extract showed stronger antibacterial activity than root extract.

Both extracts were shown to be bactericidal in their mode of action. Quercetin may contribute to

the activity of leaf extract (Mazumder et al., 2007). In another study, it was reported that crude

extract from seeds of C. ternatea L. showed maximum zone of inhibition against E. coli and

minimum with M. flavus and the callus extract showed maximum zones of inhibition against

Salmonella typhi while the lowest with E. coli and S. aureus respectively (Kelemu et al., 2004).

Alcoholic and aqueous extracts from in vitro raised calli were tested for antibacterial activity by

agar well diffusion method against Gram-negative bacteria. Antibacterial activity was shown

against Salmonella spp. and Shigella dysenteriae organisms causing enteric fever (Mhaskar et

al., 2010). In addition, the methanol crude extracts showed anti-bacterial activity against

Klebsiella pneumonia and Pseudomonas aeruginosa (Shekawat and Vijayvergia, 2010). The

crude extract from seeds of C. ternatea L. showed strong anti-fungal activity on the test fungus

Aspergillus niger and Aspergillus ochraceous followed by other organisms (Mhaskar et al.,

2010).The presence of small molecular weight, cystein rich protein, finotin obtained from seeds

of the plant C. ternatea L. has been demonstrated for its anti-fungal property (Segenet et al.,

2004). The extract which was prepared from C. ternatea L. leaves, was assessed for anti-fungal

activity against selected fungi (Aspergillus niger). The extract showed a favorable anti-fungal

activity (Kamilla et al., 2009). The crude extract from seeds of C. ternatea L. showed strong

anti-fungal activity on the test fungus Rhizoctonia solani (Segenet et al., 2004). Anti-microbial

activity of C. ternatea L. against the fish pathogens viz., Pseudomonas aeruginosa, Escherichia

coli, Klebsiella pneumonia, Bacillus subtilis, Aeromonas formican, Aeromonas hydrophila and

Streptococcus agalactiae isolated from diseased Tilapia (Oreochromis niloticus) was

investigated. Different extracts used in the study showed inhibitory effects against the fish

pathogens (Ponnusamy et al., 2010).

46

E. Proteolytic activities: Proteiolytic activities of endo-peptidases, carboxypeptidase benzyloxy

carbonyl and arylamidases lysophosphatidic acid and N-benzoyl-L-arginine P-nitro-analide of

the plant, C. ternatea L. cotyledons and axis of resting and germinating seeds was evaluated.

The study documented that the activities of carboxypeptidase and the arylamidase increased in

cotyledons reaching a maximum at the day 9, while the endopeptidases showed an increase at the

day 3 followed by adecrease. In the axial tissue the endopeptidases and carboxypeptidase

activities showed an increase until the day 9 followed by a decrease and arylamidase were low.

The increase of acidic endopeptidases and carboxypeptidase activities in germinating cotyledons

is an indication of their participation in the degradation of the storage proteins (Gupta et al.,

2010).

F. Diuretic activity: The powdered form of dried whole root and ethanol extract were evaluated

in dogs for diuretic activity and it was reported that only single intra-venous dose of extract

produced moderate increase in urinary excretion of Na, K and decrease in Cl but no change in

urine volume (Piala et al., 1962).

G. Antioxidant Activity: The DPPH free radical scavenging assay of aqueous and ethanol

extracts of C. ternatea L. was studied. The potential antioxidant activity of C. ternatea L.

extracts and an extract containing eye gel formulation was investigated. Aqueous extracts were

shown to have stronger antioxidant activity (as measured by DPPH scavenging activity) than

ethanol extracts (Kamkaen & Wilkinson, 2009). The nephro protective & antioxidant activities

of the ethanol extract of aerial parts of C. ternatea L. on acetaminophen induced toxicity in rats

was studied. Biochemical studies showed that there was an increase in the levels of serum urea

and creatinine along with an increase in the body weight and reduction in the levels of uric acid

in acetaminophen induced groups. These values retrieved significantly by treatment with C.

ternatea L. extracts at two different doses. The antioxidant studies revealed that the levels of

renal SOD, CAT, GSH and GPx in the APAP treated animals were increased significantly along

with a reduced MDA content in ethanol extract of C. ternatea L. treated groups. These data

suggested that the ethanol extract of C. ternatea L. prevented renal damage from APAP induced

nephrotoxicity in rats and it was likely to be mediated through active phyto constituents and its

antioxidant activities (Sarumathy et al., 2011). Methanol and water extracts of the leaves and

47

flowers of C. ternatea L. showed DPPH scavenging activity in a dose dependent manner (Rabeta

and Nabil, 2013)

H. Larvicidal Activities: Screening of natural products for mosquito larvicidal activity against

three major mosquito vectors Aedes aegypti, Culex quinquefasciatus and Anopheles stephensi

was done. The study resulted in the identification of three potential plant extracts viz., Saraca

asoca, Nyctanthes arbortristis, and C. ternatea L. for mosquito larval control. Among the three

plant species studied for mosquito larvicidal activity, C. ternatea L. was showing the most

promising mosquito larvicidal activity (Mathew et al., 2009).

I. Anti-diabetic Activities: Oral administration of aqueous extract of C. ternatea L. leaves and

flowers for 84 days showed significant reduction in serum glucose, glycosylated hemoglobin,

total cholesterol, triglycerides, urea, creatinine and the activity of gluconeogenic enzyme

glucose-6-phosphatase, but increase in serum insulin, HDL-cholesterol, protein, liver and

skeletal muscle glycogen content and the activity of glycolytic enzyme glucokinase (Daisy &

Rajathi, 2009). Oral administration of alcoholic extract of root of C. ternatea L. significantly

reduced blood glucose level in experimental rats (Daisy et al., 2009; Ravishankar & Jevoor,

2013).

J. Immunomodulatory Activities: This study evaluated the immune stimulatory activities of

aqueous extracts of C. ternatea L. leaf and flower. The studies were conducted on oral

administration of aqueous extract of C. ternatea L. to alloxan-induced diabetic rats for a duration

of 60 days which significantly decreased the in serum glucose and cholesterol levels. The total

white blood cells, red blood cells, T-lymphocytes and B-lymphocytes were significantly

increased in treated animals, while monocytes and eosinophils showed an opposite trend. These

results further indicate that these plant extracts have immunomodulatory effects that strengthen

the immune system (Daisy et al., 2004). Immunomodulatory activity of the plant was also

studied on male albino rats (Solanki & Jain, 2010).

K. Effect of C. ternatea L. on general behavior: Ethanol extract of the root of C. ternatea L.

was evaluated for different neuro-pharmacological actions in rats and mice, such as general

48

behavior, exploratory behavior, muscle relaxant activity and phenobarbitone induced sleeping

time. The ethanol extract caused reduction in spontaneous activity, decrease in exploratory

behavioral pattern, reduction in the muscle relaxant activity indicating significant

neuropharmacological activity (Boominathan et al., 2003).

L. Hepatoprotective Activities: Hepatoprotective effects of C. ternatea L. leaves were reported

in some investigations. In one study, this property was evaluated in rats with carbon tetrachloride

induced hepatotoxicity. It was reported that there was decreased level of serum marker enzymes

and increased total protein, total conjugated and unconjugated bilirubins indicating the

hepatoprotective effect of the plant (Shanmugasundaram et al., 2010). In another study,

hepatoprotective effect of the leaves was established against paracetamol induced damage in

mice (Nithianantham et al., 2011). In another study, hepatoprotective activity was studied using

the seeds of the plant. The rats used in the study showed decreased level of liver enzymes

proving the hepatoprotective nature (Solanki and Jain, 2011). Hypoglycemic effect was tested in

alloxan-induced diabetes in rats using aqueous extracts of C. ternatea L. leaves and flowers.

Results showed significant reduction in serum glucose, glycosylated hemoglobin and the

activities of gluconeogenic enzyme, glucose-6- phosphatase, but increase in serum insulin, liver

and skeletal muscle glycogen and the activity of the glycolytic enzyme, glucokinase (Daisy &

Rajathi, 2009). The hepatoprotective effects of the roots of white and blue colour flower variety

was studied and reported that the white colour flower variety showed more significant

hepatoprotective activity than the blue one (Patil and Patil, 2011).

M. Anti-cancer Activities: Anti-cancer activity of the plant C. ternatea L. was evaluated by a

number of workers. Anti-cancer activity was evaluated from the leaves (Ramaswamy et al.,

2011), aerial parts (Sarumathy et al., 2011) and seeds (Jacob and Latha, 2013) of the plant.

Cyclotides were isolated from C. ternatea L. flower and seeds and was evaluated for the anti-

cancer activity. It was found that the cyclotides had significant anticancer and chemosensitizing

abilities; such cyclotides were capable of causing multi-fold decreases in the half maximal

inhibitory concentration value in the presence of paclitaxel (Sen et al., 2013).

49

N. Other Activities: The plant of interest was found to be active as nitrogen supplements to

Napies grass basal diet in relation to the performance of lactating Jersey crows (Juma et al., 2006

& Marin et al., 2003). C. ternatea L. seeds were evaluated on milk induced leucocytosis and

milk induced eosinophilia in mice and found significant inhibition. The ethanol and benzene

extracts showed milk induced leucocytosis in dose dependent manner. But in milk induced

eosinophilia ethanol extracts showed inhibition eosinophilia in dose dependent manner while

benzene extract does not showed dose dependent inhibition. This inhibition of leucocytosis and

eosinophilia indicates the anti-allergic potential of C. ternatea L. (Taur & Patil, 2009).

2.3.3: Phytochemical Constituents of C. ternatea L.:

Phytochemical investigation of different parts of the plant has been done by many workers.

Phytochemical screening of the roots shows the presence of ternatins, alkaloids, flavonoids,

saponins, carbohydrates, proteins, resins, starch, taraxerol and taraxerone (Uma et al., 2009). The

major phytoconstituents found in C. ternatea L. are the pentacyclic triterpenoids such as

taraxerol and taraxerone (Banerjee & Chakravarti, 1963; 1964) (Fig-2.3). A new simple,

sensitive, selective and precise HPLC method was developed for the determination of taraxerol

in C. ternatea L., which was being performed on TLC aluminum plates (Kumar et al., 2008). A

wide range of secondary metabolites including triterpenoids, flavonol glycosides, anthocyanins

and steroids has been isolated from C. ternatea L. (Mukherjee et al., 2008). Four kaempferol

glycosides I II, III and IV were isolated from the leaves of C. ternatea L. Kaempferol-3-

glucoside (I), kaempferol-3 -rutinoside (II) and kaempferol-3-neohesperidoside (III) were

identified by ultra violet, protein magnetic resonance and mass spectrometry and Kaempferol-3-

orhamnosyl glucoside (IV) from spectral data and was named Clitorin (Morita et al., 1977). The

seeds contain nucleoprotein with its amino-acid sequence similar to insulin, delphinidin-3,3,5-

triglucoside, essential amino-acids, pentosan, watersoluble mucilage, adenosine, an anthoxanthin

glucoside, greenish yellow fixed oil (Joshi et al., 1981), a phenol glycoside, 3,5,7,4-tetrahydroxy

-flavone-3-rhamoglycoside, an alkaloid , ethyl D-galactopyranoside, p-hydroxycinnamic acid

polypeptide, a highly basic protein-finotin, a bitter acid resin, tannic acid, 6% ash and a toxic

alkaloid (Potsangbam et al., 2008). According to Yoganarasimhan, seeds contain g-sitosterol, ß-

sitosterol, and hexacosanol and anthocyanin glucoside (Yoganarasimhan, 2000 and Sinha, 1960-

b). It has been reported that a lectin present in the seeds of C. ternatea L. agglutinated trypsin-

50

treated human B erythrocytes (Naeem et al., 2007). Since the purified lectin was found to be

potential tool for cancer studies so an attempt was made for the alternate high yielding

purification method for C. ternatea L. lectin designated CTL, present in the seeds of this member

of leguminosae family (Naeem et al., 2007). Another study demonstrated that minor delphinidin

glycosides, eight anthocyanins (ternatins C1, C2, C3, C4, C5 and D3 and preternatins A3 and

C4) were isolated from the young C. ternatea L. flowers (Terahara, 1998). Another study

showed that malonylated flavonol glycosides were isolated from the petals of C. ternatea L. with

different petal colors using LC/MS/MS (Kazuma et al., 2003). It was also reported that five new

anthocyanins, ternatins A3, B3, B4, B2 and D2 were isolated from C. ternatea L. flowers

(Terahara, 1996). Ranaganayaki and Singh (1979) reported kaempferol and Saito et al. (1985)

detected kaempferol-3-glucoside, kaempferol-3-robinobioside-7-rhamnoside, quercetin and

quercetin 3-glucoside. Six ternatins A1, A2, B1, B2, D1 and D2 in C. ternatea L. flowers were

isolated by reversed phase high performance liquid chromatography and their structures were

partly characterized as highly acylated delphinidin derivatives (Terahara et al., 1990). C.

ternatea L. was powdered and evaluated quantitatively for the analysis of total soluble sugars,

protein, phenol, starch, carbohydrate and lipid (Shekhawat & Vijayvergia, 2010).

Fig-2.3: Some of the phytochemicals isolated from C. ternatea L. (from Sethiya et al., 2009).

51

2.4: SIGNIFICANCE OF THE PRESENT WORK:

The rapid growth of the world's population over the past one hundred years results from a

difference between the rate of birth and the rate of death. The growth in human population

around the world affects all people through its impact on the economy and environment. Latest

official current world population estimate is 7,021,836,029 (7 billion), where the total population

in India is 1,210,193,422 (1.2 billion) in 2011 population census. The total population of Assam

is 31,169,272, and the total population estimated in Barpeta district is 1,693,190 (Das, 2011).

While total population of India has increased from 36 crore in 1951 to 1.2 billion in 2011, the

country’s total fertility rate (TFR) has come down from 6.1 to 2.62 in last 50 years (Chaudhri

and Jha, 2011) (Fig-2.4). In Assam present TFR is 2.6, where as that of Barpeta district is 3.4

(Das, 2011). The total fertility rate is declining in progressive manner from the last decades.

Infertility varies across regions of the world and is estimated to affect 8 to 12 per cent of couples

worldwide (Sciarra, 1994). In 2010, an estimated 48.5 million couples worldwide were infertile

(Mascarenhas et al., 2012). International estimates of infertility prevalence showed 3.5% to

16.7% cases in more developed nations and from 6.9% to 9.3% in less-developed nations

(Boivin et al., 2007). In US, infertility cases are going on increasing and it has been documented

that the present prevalence of infertility is 15.5% (Thoma et al., 2013). In Sub-Saharan Africa,

the prevalence of infertility, ranged from less than 10 percent in Togo and Rwanda to about 25

percent in Cameroon and Central African Republic of women aged 20-44 years (Larsen,

2000).There are sparse data on the prevalence of primary infertility in India. The WHO estimates

the overall prevalence of primary infertility in India to be between 3.9 and 16.8 per cent (WHO,

2004). Estimates of infertility vary widely among Indian states from 3.7 per cent in Uttar

Pradesh, Himachal Pradesh and Maharashtra to 5 per cent in Andhra Pradesh (Unisa, 1999),

12.6% in Mysore (Adamson et al., 2011) and 15 per cent in Kashmir (Zargar et al., 1997).

Moreover, the prevalence of primary infertility has also been shown to vary across tribes and

castes within the same region in India (Kumar, 2007). In Assam the infertility rate was reported

to 1.4% by the United Nations Population Fund, 2003. There is no viable up-to-date data

regarding the statistics of infertility in Assam, but the rate is going on increasing day by day.

With this increase there is reduction of fertility rate is obvious. Therefore there is an urgent need

to explore viable herbal resources for curing infertility

order for enhancing the rate of fertility.

Fig-2.4: Changes in Total Fertility Rate in India from 1961 to 2011,

The introduction of IVF in 1978 created a new treatment

fertility problems. IVF employs advanced technologies to recover mature ova, fertilize them in

the laboratory, and then implant them in the uterus, thus bypassing the fallopian tubes.

Evaluating and treating infertile couple

the latest in assisted reproduction technology, such as in

US$50,000 per live birth. But success rates everywhere are limited: about 15 to 20 percent of

women have a clinical pregnancy; three quarters of these have viable births. In addition to the

high costs, centers in developing countries may have difficulty ensuring access to appropriate

facilities and supplies (Okonofua, 1996).

IVF and ICSI are more likely to be born preterm, of low birth weight and to be a twin or higher

order multiple than spontaneously conceived infants (Beral and Doyle, 1990; Jackson

2004). In addition to the ART facilities many couples take their treatments through fertility

drugs. But, the fertility drugs may cause multiple gestations, ectopic pregn

has been also reported that fertility drugs may elevate the

2002) as well as breast cancer (Fei

but due to financial and many other problems, people of Assam still go for a traditional healer as

their first choice.

52

to explore viable herbal resources for curing infertility and other reproductive abnormalities

order for enhancing the rate of fertility.

: Changes in Total Fertility Rate in India from 1961 to 2011, (Taken and seveloped from Chaudhri

and Jha, 2011).

The introduction of IVF in 1978 created a new treatment option for women suffering from



Evaluating and treating infertile couples can be a costly process. At the extreme, offering couples

the latest in assisted reproduction technology, such as in-vitro fertilization, may cost upwards of


men have a clinical pregnancy; three quarters of these have viable births. In addition to the


facilities and supplies (Okonofua, 1996). It is well established that infants conceived following


order multiple than spontaneously conceived infants (Beral and Doyle, 1990; Jackson


the fertility drugs may cause multiple gestations, ectopic pregnancies, torsion etc

has been also reported that fertility drugs may elevate the risk of ovarian cancer (Nes

2002) as well as breast cancer (Fei et al., 2012). In Assam, there are number of fertility clinics,


eproductive abnormalities in

(Taken and seveloped from Chaudhri

option for women suffering from



s can be a costly process. At the extreme, offering couples

vitro fertilization, may cost upwards of


men have a clinical pregnancy; three quarters of these have viable births. In addition to the


at infants conceived following


order multiple than spontaneously conceived infants (Beral and Doyle, 1990; Jackson et al.,


ancies, torsion etc. It

risk of ovarian cancer (Nes et al.,

., 2012). In Assam, there are number of fertility clinics,


53

The traditional approach makes use of material that has been found by trial and error over many

years in different cultures and systems of medicine. Plants that were once considered of no value

are now being investigated, evaluated and developed into drugs, with little or no side effects. A

recent review has shown that approximately 25% of modern medications have been plant

derived, while 75% of new drugs against infectious diseases that have arrived between 1981 and

2002 originated from natural sources (Bedoya, et al., 2009). The use of plant extracts as fertility

enhancer in animals is now in the increase trend because of the shifting of attention from

synthetic drugs to natural plant products (Dada & Ajilore, 2009). Fertility enhancing properties

of many plants have been evaluated by many workers in last decades. There are number of

reports for inducing male infertility and enhancement of male fertility by treating with plant

extracts, but there are a few reports on female fertility enhancers. It is very essential to explore

the traditionally used plants to enhance fertility from this part of India. Fertility drug of plant

origin which acts as fertility enhancer is an utmost need for those who are suffering from fertility

problems. This type of drug can contribute to increase the fertility rate and hence helps to

overcome the global problem of decrease of total fertility rate.

chapter ii review of literature -...

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