research article gastroprotective effect of freeze dried stripped...

Research ArticleGastroprotective Effect of Freeze Dried Stripped SnakeheadFish (Channa striata Bloch) Aqueous Extract against AspirinInduced Ulcerogenesis in Pylorus Ligated Rats

Mohammed Safwan Ali Khan12 Abdul Manan Mat Jais1 Javeed Hussain2 Faiza Siddiqua2

A Gopala Reddy3 P Shivakumar3 and D Madhuri4

1 Pharmacology amp Toxicology Laboratory Department of Biomedical Sciences Faculty of Medicine and Health SciencesUniversity Putra Malaysia 43400 Serdang Selangor Darul Ehsan Malaysia

2 Natural Products Research Laboratory and Department of Pharmacology Anwarul Uloom College of PharmacyNew Mallepally Hyderabad Andhra Pradesh 500 001 India

3 Department of Pharmacology amp Toxicology College of Veterinary Science Acharya NG Ranga Agricultural UniversityRajendra Nagar Hyderabad Andhra Pradesh 500 030 India

4Department of Pathology College of Veterinary Science Acharya NG Ranga Agricultural University Rajendra NagarHyderabad Andhra Pradesh 500 030 India

Correspondence should be addressed to Mohammed Safwan Ali Khan safwanpharmagmailcom

Received 8 January 2014 Accepted 2 March 2014 Published 29 May 2014

Academic Editors H Cerecetto V Manju and S Mingmalairak

Copyright copy 2014 Mohammed Safwan Ali Khan et al This is an open access article distributed under the Creative CommonsAttribution License which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited

Channa striata (Bloch) is a fresh water fish belonging to the family ChannidaeThe stripped snakehead fish possesses wide range ofmedicinal properties In view of traditional use of C striata for wound healing the present study was undertaken to investigate thebeneficial effects of orally administered freeze dried aqueous extract of Channa striata (AECS) in experimentally induced gastriculcers in Wistar rats Aspirin induced ulcerogenesis in pyloric ligation model was used for the assessment of antiulcer activity andRanitidine (50mgkg) was employed as the standard drugThe various gastric parameters like volume of gastric juice pH free andtotal acidities ulcer index and levels of antioxidant enzymes like catalase superoxide dismutase and lipid peroxidation markermalondialdehyde were determined AECS at concentrations of 40 and 50wv significantly decreased the volume of gastric juiceand increased the levels of catalase while considerable decrease in free and total acidities and increase in superoxide dismutase wereobserved with the treatment of standard drug and AECS (50wv) All the test doses of AECSmarkedly decreased ulcer index andmalondialdehyde compared to the standard drug whereas AECS 30 wv did not alter volume of gastric juice pH free and totalacidities catalase and superoxide dismutase From these findings it can be concluded that AECS is devoid of acid neutralizingeffects at lower doses and possesses antisecretory and antiulcer activities and this could be related to its antioxidant mechanism

1 Introduction

Channa striata (Bloch) is commonly known as strippedsnakehead fish and Haruan in Malay C striata is indigenousto Malaysia It is a tropical and air breathing carnivorousfreshwater fish from the family Channidae There are alltogether 30 species in the family reported around the worldand eight of them are found in Malaysia Members ofthe family are also found in all ASEAN countries namely

MyanmarThailand Laos Cambodia Vietnam Brunei Phil-ippines Indonesia and Singapore [1]C striata is wild speciesfound in small rivers lakes pools and shallow water bodiesand in its natural habitat It can survive in harsh environmentswith low dissolved oxygen and high ammonia [2] C striatacontains all the essential amino and fatty acids required forwound healing [3 4] the fatty acid content of this fish isrich in arachidonic acid (ARA) [5] Mohd Shafri and MatJais reviewed in detail the description habitat traditional

Hindawi Publishing CorporationISRN PharmacologyVolume 2014 Article ID 327606 8 pageshttpdxdoiorg1011552014327606

2 ISRN Pharmacology

therapeutic uses chemical composition and scientificallyproven medicinal properties [6] The fish is reported topossess antinociceptive [7 8] antipyretic [9] antidepressant[10 11] anti-inflammatory [12] antifungal [13] antimicrobial[14] antiosteoarthritic [15] neuroregenerative and restora-tive [16] properties and enhances wound healing process [17ndash19] In view of traditional and scientific reports on woundhealing potential of C striata and presence of wide range ofbioactive components the present study was undertaken toinvestigate antiulcer activity

2 Material and Methods

21 Preparation of Aqueous Extract of Channa striata (Bloch)and Freeze Drying AdultChanna striata (Bloch) of mediumsize (total length of fish 2060ndash4410 cm) weighing 250ndash400 gfish were collected in November 2011 from KualaTerengganu Terengganu District (N 5∘191015840294010158401015840 and E103∘081015840272910158401015840) from Peninsular Malaysia The temperaturewas 28ndash33∘C with pH between 560 and 820 salinity 0 pptdissolved oxygen in the range of 200ndash1100mgL turbidityin the range of 100ndash1800NTU and conductivity 054-055mScmThe fishes were verified by Terengganu FisheriesDepartment Ministry of Agriculture Malaysia Fillets wereobtained by carefully cutting the fish lengthwise along back-bone to gain maximum amount of flesh The whole fish filletextract was prepared by using pressure cooker set at 100∘Cfor 30min The fresh boneless fillet was weighed and placedon a stainless steel wire mesh mounted on a stainless steeltripod in the pressure cooker Fish fillet and distilled waterwere added in volume ratio of 1 2 The extract was obtainedthrough steaming At the end of the extraction procedurethe fillet was discarded while collecting the liquid extractusing Whatman no 1 filter paper The obtained extract wasfreeze dried using a freeze drier and stored at 20∘C until use[7 8]

22 Drugs and Chemicals Chloroform AR phenolphthaleinpH indicator solution and sodium hydroxide pellets wereprocured from SD Fine-Chem Limited Mumbai while pureaspirin was obtained from Divis Laboratories HyderabadRanitidine (as Rantac 150mg) was from JB Chemicals andPharmaceuticals Mumbai while surgical spirit was obtainedfrom Kakatiya Pharma Hyderabad Topferrsquos reagent anddistilled water were obtained from Nice Chemicals Chinaand Stangen Fine Chemicals Hyderabad respectively Allchemicals were used without further purification

23 Animals For the evaluation of the antiulcer propertymale albino Wistar rats weighing 160ndash200 g were usedThe study was conducted after the approval of protocol byInstitutional Animal Ethical Committee (IAEC)The animalswere maintained under standard conditions that is housedin polypropylene cages and maintained at a temperature27 plusmn 2

∘C relative humidity 65 plusmn 10 under 12-hour lightand dark cycle The animals were fed with standard pelletdiet with water ad libitum in an animal house (Reg num-ber 1534POa11CPCSEA) approved by the Committee for

the Purpose of Control and Supervision on ExperimentalAnimals (CPCSEA) The animals were acclimatized for tendays under laboratory conditions before carrying out theexperiments

24 Acute Toxicity and Selection of Test Doses Earlier investi-gations indicate that Channa striata (Bloch) aqueous extractis safe and nontoxic even at the dose of 8 gkg or 100 wvconcentration [7 8] Doses of 30 40 and 50 wv wereselected for the screening of antiulcer activity based on thereport of Saleem et al [10]

25 Evaluation of Antiulcer Activity

251 Aspirin Induced Ulcerogenesis in Pylorus Ligated RatsAspirin induced ulcerogenesis in pylorus ligated rats modelwas used for the evaluation of antiulcer activity with slightmodification The animals were divided into five groups (119899 =6) Group I served as negative control and received only vehi-cle Groups II III and IV received aqueous extract ofChannastriata (AECS) at 30 40 and 50 wv concentrationsrespectively orally at the volume of 10mLkgGroupV servedas standard and was treated with standard drug (Ranitidine50mgkg po) [20] Aspirin suspended in 1 CMC in waterwas administered orally at a dose of 500mgkg in 12-hourfasted rats [21] The test extract and standard drug treatmentwere done 30min prior to the administration of aspirin After30min the pyloric ligation surgery was performed Fourhours later the animals were sacrificed by euthanasia

Collection and Measurement of Gastric Juice The stomachswere excised carefully keeping the esophagus closed Thestomachs were opened along the greater curvature removingthe luminal contents The gastric contents were collected andcentrifuged at 1000 rpm for 10 minutes After centrifugationsamples were decanted and the volume of gastric juice wasnoted and is expressed asmL100 g bodyweightThe contentswere subjected to analysis for free and total acidities

Determination of Gastric Juice pH One mL of supernatantliquid was diluted to 10mL with distilled water The pH ofthe solution was recorded with the help of digital pH meter

Estimation of Total and Free Acidities The above solutionwas titrated against 001N NaOH using Topferrsquos reagent asindicatorThe endpoint of the titration was when the solutionturns orange in colour The volume of NaOH was notedwhich corresponds to the free acidity Further the titrationwas continued till the solution regained pink colour Thetotal volume of NaOH was noted which corresponds to totalacidity

Determination of Ulcer Index Mean ulcer score for eachanimal is expressed as ulcer indexThe stomachswerewashedwith running water to see the ulcers in the glandular portionof the stomachThe number of ulcers per stomach was notedand the severity of the ulcers was scoredmicroscopically with

ISRN Pharmacology 3

the help of hand lens (10x) and scoring was done as per Asru[22]

0 = Normal stomach05 = Red coloration1 = Spot ulcers15 = Haemorrhagic streaks2 = Ulcers gt3mm but lt5mm3 = Ulcers gt5mm

The percentage of protection was calculated by the formula

Percentage protection = 100 minus (UtUctimes 100) (1)

where Ut = ulcer index of the treated group and Uc = ulcerindex of control group

Assessment of Oxidative Damage in Gastric Tissue Aftermeasuring the ulcer index the stomachs were washed with09 (wv) NaCl cut into small pieces and homogenizedwith a glass homogenizer in ice-cold 015M KCl solution toproduce a 20 (wv) homogenateThe homogenate was usedfor the determination of various biochemical parameters

(1) Estimation of Catalase Catalase containing sample isallowed to split H

2O2followed by adding dichromateacetic

acid mixture to stop the reaction Dichromate in acetic acidis reduced first to unstable blue colored perchromic acid andfinally to stable green colored chromic acetate in the presenceof H2O2 which is measured colorimetrically at 570 nmThus

chromic acetatemeasured gives the amount of freely availableH2O2 To assay mixture containing 04mL of 02M H

2O2

and 05mL of 001M phosphate buffer (pH 7) 01mL ofhomogenate was added and mixed well Into this 2mL ofdichromate acetic acid solution was blown exactly after 1minand kept in boiling water bath for 10min The absorbanceof green colored chromic acetate formed was measured at570 nm against reagent blank containing 04mL of 02MH2O2and 05mL of 001M phosphate buffer (pH 7) The

enzyme level is expressed as unitsmg of tissue [23]

(2) Estimation of Superoxide DismutaseThe reaction involvesgeneration of superoxide by pyrogallol auto-oxidation andthe inhibition of superoxide dependent reduction of thetetrazolium dye MTT [3-(4-5 dimethyl thiazol 2-yl) 25-diphenyl tetrazolium bromide] to its formazan The reactionis terminated by the addition of dimethyl sulfoxide (DMSO)which helps to solubilize the formazan formed The colorevolved is stable formanyhours and is expressed as SODunits(one unit of SOD is the amount in mg of protein required toinhibit theMTT reduction by 50)The reagents were addedin the sample control and the blank as shown in Table 1

The absorbance was read at 570 nm against distilledwater (blank) Superoxide dismutase was expressed as SODunitsmg of tissue [24]

(3) Estimation ofMalondialdehydeMalondialdehyde formedfrom the breakdown of polyunsaturated fatty acids serves as

Table 1 Details of various reagents and their quantities added tosample control and blank during the estimation of superoxidedismutase

Reagents Sample Control Blank(duplicate)

PBS 065mL 065mL 065mLMTT 30 120583L 30120583L 30 120583LHomogenate 10 120583L mdash mdashPyrogallol 75 120583L 75 120583L 75 120583LThe sample control and blank were incubated for 5min at roomtemperatureDMSO 075 075 075

a convenient index for determining the extent of peroxidationreactionMalondialdehyde has been identified as the productof lipid peroxidation that reacts with thiobarbituric acid togive a red colour absorbing light maximally at 535 nm One gof tissue sample with 10mL of 02M Tris HCl buffer (pH 72)was taken in a tissue homogenizer to get a 10 homogenate500120583L of supernatant from the homogenate 1mL of 10trichloroacetic acid and 1mL of 067 thiobarbituric acidwere taken in a tightly stoppered tube The tube was heatedto boiling temperature for 45min After cooling the tubethe contents were centrifuged The supernatant was read at532 nm against blank The concentration of test samples wasobtained using molar extinction coefficient of MDA Theamount ofMDA is expressed as number ofmoles ofMDAmgof tissue [25]

26 Statistical Analysis Data obtained was analyzed by one-way ANOVA followed by Dunnettrsquos multiple comparisonspost hoc test using Graphpad Prism version 40 32 bits forwindows Graphpad software San Diego California USA(httpwwwgraphpadcom) The values are expressed asmean plusmn standard error of mean (SEM) 119875 lt 005 wasconsidered statistically significant

3 Results

31 Results of Aspirin Induced Ulcerogenesis in PylorusLigated Rats (Figure 2 and Table 2)

311 Volume of Gastric Juice AECS significantly (119875 lt 005)decreased the volume of gastric juice at the dose of 40 wvconcentration Further decrease in the output of gastric juicewas observedwith the treatment of higher concentration thatis 50wv (119875 lt 001) However the least test dose (30wv)used in the study did not show any significant antisecretoryeffect (as shown in Figure 2 and Table 2)

312 pH of Gastric Juice Treatment with the standard drug(Ranitidine 50mgkg) significantly (119875 lt 001) raised the pHfrom 383 plusmn 010 (negative control) to 495 plusmn 016 while allthe concentrations of AECS failed to elevate pH of the gastricjuice (as shown in Figure 2 and Table 2)

4 ISRN Pharmacology

Table 2 Results of aspirin induced ulcerogenesis in pyloric ligation model

ParametersVarious groups

Negative control Ranitidine Aqueous extract of Channa striata50mgkg 30 wv 40 wv 50 wv

Volume of gastric juice (mL) 543 plusmn 022 436 plusmn 017lowastlowast

526 plusmn 016ns

453 plusmn 016lowast

423 plusmn 032lowastlowast

pH 383 plusmn 0108 495 plusmn 016lowastlowast

333 plusmn 033ns

368 plusmn 018ns365 plusmn 0084

ns

Free acidity 5100 plusmn 285 4000 plusmn 247lowast


ns3933 plusmn 297

lowast

Total acidity 9317 plusmn 186 8550 plusmn 140lowastlowast


ns8667 plusmn 120

lowast

Ulcer index 591 plusmn 045 275 plusmn 038lowastlowastlowast



308 plusmn 035lowastlowastlowast

Percentage of inhibition mdash 5346 3231 3654 4788Catalase (unitsmg of tissue) 0011 plusmn 0011 0059 plusmn 00032

lowastlowast

0041 plusmn 00079ns0066 plusmn 00064

lowastlowast


Superoxide dismutase (unitsmg of tissue) 0015 plusmn 0010 0071 plusmn 00094lowastlowast 0027 plusmn 00050ns 0047 plusmn 0012ns 0055 plusmn 0013lowast

Malondialdehyde (molesmg of tissue) 0190 plusmn 00012 0089 plusmn 00013lowastlowastlowast




Note sample size (119899) = 6 rats per group Data is expressed as mean plusmn standard error of mean and standard deviation in parenthesis lowast119875 lt 005 lowastlowast119875 lt 001lowastlowastlowast

119875 lt 0001 and ns nonsignificant versus negative control (on statistical analysis with ANOVA followed by Dunnettrsquos multiple comparison post hoc test)

313 Free and Total Acidities Likewise 30 and 40 wvsolutions of AECS did not alter free and total aciditiesA slight (119875 lt 005) decrease in free and total aciditieswas observed with the treatment of 50 wv AECS solutionagainst the significant decrease in free and total acidities bythe standard drug with 119875 lt 001 and 119875 lt 0001 respectively(as shown in Figure 2 and Table 2)

314 Ulcer Index (UI) All the test doses of AECS solutions(30 40 and 50 wv) significantly decreased the ulcerindex (119875 value ranging from 001 to 0001) The decrease inulcer index in the AECS (50 wv) treated group is compa-rable with that of the standard drug A dose related increasein percentage protection was observed The percentage ofinhibition for 40 and 50 AECS was found to be 3654and 4788 respectively while the standard showed 5346(as shown in Figure 2 and Table 2)

315 Catalase (CAT) A dose dependent increase wasnoticed with the treatment of AECS with 119875 value rangingfrom 001 to 0001 except the lowest test dose used in theexperiment The results of standard and 40 wv AECS werefound to be similar while 50 AECS exhibited maximumincrease in the level of CAT with 119875 lt 0001 (as shown inFigure 2 and Table 2)

316 Superoxide Dismutase (SOD) AECS at 30 and 40wv concentrations did not cause significant increase in thelevel of superoxide dismutasewhilst 50wvAECS raised thelevel of SOD with 119875 lt 005 The treatment with the standarddisplayed the most significant rise with 119875 lt 001 (as shownin Figure 2 and Table 2)

317Malondialdehyde (MDA) The standard drug and all thetest doses of AECS significantly decreased the formation ofmalondialdehyde an end product of lipid peroxidation Apotent as well as consistent decrease in the level of MDA(119875 lt 0001) was observed The decrease in MDA wasproportionate to the increasing test doses further the effect

of all the concentrations of AECS was more profound thanthe standard This highlights the gastroprotective potentialof AECS against lipid peroxidation of gastric mucosa Theresults are displayed in Figure 2 and Table 2

318 Macroscopy of Stomachs

(a) Negative Control The stomach shows red colorationhaemorrhage hyperaemia one spot ulcer two ulcerswith diameter in range of 3ndash5mm and two ulcersgreater than 5mm

(b) AECS (30 wv) Red coloration and four spot ulcersare seen in the stomach

(c) AECS (40 wv) The stomach coloration is normalbut shows two spot ulcers

(d) AECS (50 wv) Erythema an inflammatory signcan be observed in the stomach with completeabsence of ulcers

(e) Standard The stomach reveals partial red colorationwith no ulcers The images are shown in Figure 1

4 Discussion

Channa striata contains unsaturated fatty acids (UFA) andessential amino acids (AA) that stimulate and promotehealing of wounds [3 18 19 26] C striata contains alaninearginine aspartic acid glutamic acid glycine leucine andproline [27] An earlier report of Gam et al reported lysinethreonine and valine as the othermost abundant amino acidsin C striata [28] Some of these amino acids are the integralcomponents of gastric mucous [29] Furthermore some ofthese amino acids are known to have significant antioxidantproperties particularly with linoleic acid [30]

C striata fresh fillet also contains high amount of glycinewhich is one of the important components responsible for theformation of collagen in the various tissues of human body [327 31] C striata treatment promotes remodeling of collagenthrough the synthesis of inter- and intramolecular protein

ISRN Pharmacology 5

(a) (b) (c) (d) (e)

Figure 1 Photos of ratrsquos stomachs subjected to aspirin induced ulcerogenesis in pyloric ligation model Note (a) negative control (b) Test-I(AECS 30 wv) (c) Test-II (AECS 40 wv) (d) Test-III (AECS 50 wv) and (e) standard (Ranitidine 50mgkg)

cross-linking This action strengthens the body tissues andprevents further degradation [1 15] Glycine is the aminoacid present in C striata in the highest concentrationGlycine is important in healing process as it is one of themajor components of human tissue collagen It promotestissue repair synergistically by forming a polypeptide withother essential amino acids like alanine arginine isoleucinephenylalanine proline and serine [32]

Arginine plays critical and multiple roles in woundhealing process It stimulates the release of hormones likegrowth hormone from pituitary and insulin from pancreas[33] In postinjury catabolic state it decreases the urinarynitrogen loss to regulate nitrogen balance [34] This aminoacid is also a substrate for two integral enzymesmdashnitric oxidesynthetase (NOS) and arginase Arginine is metabolized inwounds by the action of enzyme arginase abundantly presentin wound fluid [35] Arginine produces hydroxyproline animportant component (91 of the total amino acid residue)of collagen [36] Aspartic acid an excitatory amino acid thatis involved in antioxidant mechanism is also found in highamount in C striata [37]

The dominant fatty acids in C striata are palmitic acid(C16 0) stearic acid (C18 0) oleic acid (C18 1n-9) andlinoleic acid (C18 2n-6) [27] Polyunsaturated fatty acids(PUFA) components (arachidonic and linoleic acids) of Cstriata decrease serum cholesterol and triglycerides levelsand inhibit clotting of blood [38] This action can facilitatethe supply of blood to the gastric tissue countering theobstruction one of the etiological factors in the pathogenesisof peptic ulcers Further the report of Mat Jais et al suggeststhe high compositions of fatty acids and certain aminoacids as responsible agents for the healing effects of Cstriata [3] The oleic and stearic acids are reported to atten-uate polymorphonuclear leukocytes activity and influencemembrane fluidity consequently suppressing inflammatoryprocesses [27] Arachidonic acid which is a precursor ofprostaglandin is found in C striata in considerable amountsThe prostaglandins play a major role in growth of tissue andwound healing [39ndash41]

Furthermore Huang et al have reported that arachi-donylglycine (a lipoamino acid formed by the conjugation

of arachidonic acid and glycine) suppresses edema andpain [42] This can relieve or minimize the gastric distressthat accompanies peptic ulceration Vitamin-A (Retinol) anessential factor for wound healing is also present in highconcentration in C striata [3 43]

C striata extract has also demonstrated inhibitory effectsonH pylori [44] Besides promising results as an antibacterialand antifungal agent against certain strainsC striata extractsexhibited antibacterial activity in various studies against widerange of bacteria like Staphylococcus aureus [13] Aeromonashydrophila andPseudomonas aeruginosa [14]Bacillus subtilisKlebsiella pneumoniae and Pseudomonas aeruginosa [45] Cstriata extracts also demonstrated antifungal activities againstAleurisma keratinophilum Botrytis pyramidalis Cordycepsmilitaris Neurospora crassa and Paecilomyces fumosoroseus[13]

The beneficial effects of aqueous extract of Channastriata (AECS) were observed at the higher test doses Theineffectiveness of low dose of AECS (30 wv) could bedue to the presence of bioactive compounds in quantitieslesser than the minimum effective dose Therefore there is aneed for a similar study at higher concentrations to evaluategastroprotective potential of AECS appropriately and justifythe observations of the present investigation properly Cstriata is a nutraceutical agent and well tolerable even atthe higher concentrations the prospect of increasing the testdoses in the future studies does exist The test doses canbe increased up to 100 wv concentration [8] or 8 gkg[7] as used by them in their studies Additionally in viewof report by Dahlan-Daud et al [27] on the chemicalcomposition of various fractions of C striata it is expectedthat a similar study using the other bioactive fractions likeHaruan Commercial Essence and Lower Phase of HaruanTraditional Extract would give more meaningful results

5 Conclusion

The present study reveals that AECS is devoid of gastricacid neutralizing effect but possesses potent antisecretory andantiulcer properties The observed pharmacological action

6 ISRN Pharmacology

8

7

6

5

4

3

2

1

0

nsVo

lum

e of g

astr

ic ju

ice (

mL)

lowast lowastlowast lowastlowast

Volume of gastric juice in various treated groups

(a)

6

4

2

0

ns ns ns

lowastlowast

pH of gastric juice in various treated groups

pH ra

nge

(b)

60

40

20

0

nsns

Volu

me o

f NaO

H (m

L) re

quire

dfo

r neu

tral

izin

g ga

stric

HCl

lowast lowast

Free acidity in various treated groups

(c)

ns nslowast lowastlowast

Total acidity in various treated groups

Volu

me o

f NaO

H (m

L) re

quire

dfo

r neu

tral

izin

g ga

stric

HCl

100

80

60

40

20

0

(d)

8

6

4

2

0

lowastlowast

lowastlowastlowastlowastlowastlowast

lowastlowast

Num

ber o

f gas

tric

ulc

ers

Ulcer index in various treated groups

(e)

ns

lowastlowast

lowastlowast

lowastlowastlowast

010

008

006

004

002

000

Level of catalase in various treated groups

Leve

l of c

atal

ase (

units

mg

of ti

ssue

)

(f)

ns

ns

lowast

lowastlowast

010

008

006

004

002

000

Leve

l of s

uper

oxid

e dism

utas

e(u

nits

mg

of ti

ssue

)

Level of superoxide dismutase in various treated groups

AECS 30wvAECS 40wv

AECS 50wvNegative controlStandard

(g)

025

020

015

010

005

000

Level of malondialdehyde in various treated groups

Leve

l of m

alon

dial

dehy

de (n

umbe

r of

mol

es o

f MD

Am

g of

tiss

ue)

AECS 30wvAECS 40wv


lowastlowastlowast lowastlowastlowastlowastlowastlowast lowastlowastlowast

(h)

Figure 2 Graphs showing effect of AECS and standard drug on various gastric parameters

ISRN Pharmacology 7

can be attributed to the essential amino acids and unsaturatedfatty acids present in the extract Further this is the firststudy that highlights antioxidant potential of AECS in an invivo experiment and indicates antioxidant activity as one ofthe probable mechanisms for the gastroprotective effect ofAECS It can be concluded that AECS possesses antisecretoryand antiulcer properties particularly at 40 and 50 wvconcentrations

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The authors acknowledge the facilities provided in carryingout the biochemical assays at the laboratory of Department ofPharmacology College of Veterinary Science Acharya NGRanga Agricultural University Rajendra Nagar Hyderabad

References

[1] AMMat Jais Y M Dambisya and T L Lee ldquoAnti-nociceptiveactivity of Channa striatus (Haruan) extract in micerdquo Journal ofEthnopharmacology vol 57 pp 125ndash130 1997

[2] KMarimuthu andM A Haniffa ldquoEmbryonic and larval devel-opment of the striped snakehead Channa striatusrdquo Taiwaniavol 52 no 1 pp 84ndash92 2007

[3] A M Mat Jais R McCulloch and K Croft ldquoFatty acid andamino acid composition in Haruan as a potential role in woundhealingrdquo General Pharmacology vol 25 no 5 pp 947ndash9501994

[4] AMMat JaisM FMatori P Kittakoop andK SowanboriruxldquoFatty acid compositions in mucus and roe of Haruan Channastriatus for wound healingrdquo General Pharmacology vol 30 no4 pp 561ndash563 1998

[5] A Zuraini M N Somchit M H Solihah et al ldquoFatty acid andamino acid composition of three local Malaysian Channa sppfishrdquo Food Chemistry vol 97 no 4 pp 674ndash678 2006

[6] M A Mohd Shafri and A M Mat Jais ldquoTherapeutic potentialof the Haruan (Channa striatus) from food to medicinal usesrdquoMalaysian Journal of Nutrition vol 18 no 1 pp 125ndash136 2012

[7] Z A Zakaria M R Sulaiman A M Mat Jais and M NSomchit ldquoEffect of various antagonists on the Channa striatusfillet extract antinociception in micerdquo Canadian Journal ofPhysiology and Pharmacology vol 83 no 7 pp 635ndash642 2005

[8] Z A Zakaria M N Somchit M R Sulaiman A M Mat Jaisand D A Israf ldquoNon-opioid antinociceptive activity of freshHaruan (Channa striatus) fillet extractrdquo Journal of Technologyand Management vol 2 pp 6ndash11 2004

[9] Z A Zakaria G H Kumar A M Mat Jais M R Sulaimanand M N Somchit ldquoAntinociceptive antiinflammatory andantipyretic properties of Channa striatus fillet aqueous andlipid-based extracts in ratsrdquo Methods and Findings in Experi-mental and Clinical Pharmacology vol 30 no 5 pp 355ndash3622008

[10] A M Saleem M T Hidayat A M Mat Jais et al ldquoAntide-pressant-like effect of aqueous extract of Channa striatus fillet

inmicemodels of depressionrdquoEuropean Review forMedical andPharmacological Sciences vol 15 no 7 pp 795ndash802 2011

[11] A M Saleem M Taufik Hidayat A M Mat Jais et alldquoInvolvement of monoaminergic system in the antidepressant-like effect of aqueous extract of Channa striatus in micerdquoEuropean Review for Medical and Pharmacological Sciences vol17 no 15 2019

[12] M N Somchit M H Solihah D A Israf Z Ahmad AK Arifah and A M Mat Jais ldquoAnti-inflammatory activityof Channa striatus Channa micropeltes and Channa luciusextracts chronic inflammatorymodulationrdquo Journal of OrientalPharmacy and Experimental Medicine vol 4 no 2 pp 91ndash942004

[13] AMMat Jais Z A Zakaria A Luo andY X Song ldquoAntifungalactivity of Channa striatus (Haruan) crude extractsrdquo Interna-tional Journal of TropicalMedicine vol 3 no 3 pp 43ndash48 2008

[14] M Dhanaraj M A Haniffa S V A Singh C M Ramakrish-nan D Manikandaraja andM J Milton ldquoAntibacterial activityof skin and intestinal mucus of five different freshwater fishspecies VizC striatesC micropeltesC maruliusC Punctatusand C gachuardquo Malaysian Journal of Science vol 28 no 3 pp257ndash262 2009

[15] N Y TMichelle G Shanthi andMY Loqman ldquoEffect of orallyadministered Channa striatus extracts against experimentally-induced osteoarthritis in rabbitsrdquo International Journal ofApplied Research and Veterinary Medicine vol 2 no 3 pp 171ndash175 2004

[16] MAMohd Shafri AMMat Jais andMKKyu ldquoNeuroregen-erative properties of Haruan (Channa striatus spp) traditionalextractrdquo Jurnal Intelek vol 6 no 1 pp 77ndash83 2011

[17] K L Wee ldquoSnakeheads their biology and culturerdquo in RecentAdvances in Aquaculture R Muir and R Roberts Eds pp 181ndash213 Westview Press Boulder 1982

[18] S H Baie and K A Sheikh ldquoThe wound healing propertiesof Channa striatus-cetrimide cream-tensile strength measure-mentrdquo Journal of Ethnopharmacology vol 71 no 1-2 pp 93ndash1002000

[19] S H Baie and K A Sheikh ldquoThe wound healing properties ofChanna striatus-cetrimide cream-wound contraction and gly-cosaminoglycan measurementrdquo Journal of Ethnopharmacologyvol 73 no 1-2 pp 15ndash30 2000

[20] R K Goel A Chakrabarti and A K Sanyal ldquoThe effect ofbiological variables on the anti-ulcerogenic effect of vegetableplantain bananardquo Planta Medica vol 2 pp 85ndash88 1985

[21] N Kannappan S Jaikumar R Manavalan and A K MuthuldquoAntiulcer activity of methanolic extract of Jatropha curcas(Linn) on aspirin-induced gastric lesions in wistar ratsrdquo Phar-macologyonline vol 1 pp 279ndash293 2008

[22] K S Asru Handbook of Experimental Pharmacology VallabhPrakashan New Delhi India 3rd edition 1999

[23] A K Sinha ldquoColorimetric assay of catalaserdquo Analytical Bio-chemistry vol 47 no 2 pp 389ndash394 1972

[24] M Madesh and K A Balasubramanian ldquoMicrotiter plate assayfor superoxide dismutase usingMTT reduction by superoxiderdquoIndian Journal of Biochemistry and Biophysics vol 35 no 3 pp184ndash188 1998

[25] K A Balasubramanian M Manohar and V I MathanldquoAn unidentified inhibitor of lipid peroxidation in intestinalmucosardquoBiochimica et Biophysica Acta vol 962 no 1 pp 51ndash581988

8 ISRN Pharmacology

[26] J-H Chyun and P Griminger ldquoImprovement of nitrogenretention by arginine and glycine supplementation and itsrelation to collagen synthesis in traumatized mature and agedratsrdquo Journal of Nutrition vol 114 no 9 pp 1705ndash1715 1984

[27] C K Dahlan-Daud A M Mat Jais Z Ahmad A Md Akimand A Adam ldquoAmino and fatty acid compositions in Haruantraditional extract (HTE)rdquoBoletin Latinoamericano y del Caribede Plantas Medicinales y Aromaticas vol 9 no 5 pp 414ndash4292010

[28] L H Gam C Y Leow and S Baie ldquoAmino acid compositionof snakehead fish (Channa striatus) of various sizes obtained atdifferent times of the yearrdquoMalaysian Journal of PharmaceuticalSciences vol 3 no 2 pp 19ndash30 2005

[29] J Schrager ldquoThe composition and some structural features ofthe principal gastric glycoproteinrdquo Digestion vol 2 no 2 pp73ndash89 1969

[30] HOHultin ldquoLipid oxidation in fishmusclerdquo inAdvances in SeaFoodBiochemistry Composition andQuality G J Flick andR EMartin Eds pp 99ndash122 Technomic Lancaster Pennysylvania1992

[31] Z A Zakaria A M Mat Jais Y M Goh M R Sulaimanand M N Somchit ldquoAmino acid and fatty acid composition ofan aqueous extract of Channa striatus (Haruan) that exhibitsantinociceptive activityrdquo Clinical and Experimental Pharmacol-ogy and Physiology vol 34 no 3 pp 198ndash204 2007

[32] M B Witte F J Thornton U Tantry and A Barbul ldquoL-arginine supplementation enhances diabetic wound healinginvolvement of the nitric oxide synthase and arginase path-waysrdquoMetabolism Clinical and Experimental vol 51 no 10 pp1269ndash1273 2002

[33] A Barbul ldquoArginine enhances wound healing and lymphocyteimmune responses in humansrdquo Surgery vol 108 no 2 pp 331ndash337 1990

[34] J Elsair ldquoEffect of arginine chlorhydrate on nitrogen balanceduring the three days following routine surgery in manrdquoBiomedicine vol 29 no 9-10 pp 312ndash317 1978

[35] J E Albina C D Mills and W L Henry Jr ldquoTemporalexpression of different pathways of l-arginine metabolism inhealing woundsrdquo Journal of Immunology vol 144 no 10 pp3877ndash3880 1990

[36] T M Devlin Textbook of Biochemistry and Clinical CorrelationWiley-Liss New York NY USA 1992

[37] C Salvatore P R Dennis P C Achille and S DanielaldquoApproach in shock inflammation and ischaemiareperfusioninjuryrdquoTheAmerican Society for Pharmacology and Experimen-tal TherapeuticsmdashPharmacological Review vol 53 pp 135ndash1592001

[38] R G Ackman ldquoNutritional composition of fats in seafoodsrdquoProgress in Food and Nutrition Science vol 13 no 3-4 pp 161ndash241 1989

[39] M P A Muntaziana S M N Amin M S Kamarudin and AA Rahim ldquoEffect of selected diets on the growth and survivalof snakehead fish (Channa striatus) fryrdquo Journal of Fisheries andAquatic Sciences vol 2013 pp 1ndash7 2013

[40] W C Bowman and M J Rand Textbook of PharmacologyBlackwell Science London UK 2nd edition 1980

[41] B G Katzung Basic and Clinical Pharmacology Appleton andLange Stamford Conn USA 6th edition 1995

[42] S M Huang T Bisogno T J Petros et al ldquoIdentification ofa new class of molecules the arachidonyl amino acids andcharacterization of one member that inhibits painrdquoThe Journalof Biological Chemistry vol 276 no 46 pp 42639ndash42644 2001

[43] S WestabyWound Healing TheMac Millan Press 2nd edition1985

[44] A M Mohamed Potential of antimicrobial activity of Channastriatus Bloch (Haruan) fillet extracts [MS thesis] UniversitiPutra Malaysia 2012

[45] OYWei R Xavier andKMarimuthu ldquoScreening of antibacte-rial activity of mucus extract of snakehead fish Channa striatus(Bloch)rdquo European Review for Medical and PharmacologicalSciences vol 14 no 8 pp 675ndash681 2010

Submit your manuscripts athttpwwwhindawicom

PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom

Volume 2014

ToxinsJournal of

VaccinesJournal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

AntibioticsInternational Journal of

ToxicologyJournal of


StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Drug DeliveryJournal of



Advances in Pharmacological Sciences

Tropical MedicineJournal of


Medicinal ChemistryInternational Journal of


AddictionJournal of



BioMed Research International

Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Autoimmune Diseases


Anesthesiology Research and Practice

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of


Pharmaceutics


MEDIATORSINFLAMMATION

of

2 ISRN Pharmacology

therapeutic uses chemical composition and scientificallyproven medicinal properties [6] The fish is reported topossess antinociceptive [7 8] antipyretic [9] antidepressant[10 11] anti-inflammatory [12] antifungal [13] antimicrobial[14] antiosteoarthritic [15] neuroregenerative and restora-tive [16] properties and enhances wound healing process [17ndash19] In view of traditional and scientific reports on woundhealing potential of C striata and presence of wide range ofbioactive components the present study was undertaken toinvestigate antiulcer activity

2 Material and Methods

21 Preparation of Aqueous Extract of Channa striata (Bloch)and Freeze Drying AdultChanna striata (Bloch) of mediumsize (total length of fish 2060ndash4410 cm) weighing 250ndash400 gfish were collected in November 2011 from KualaTerengganu Terengganu District (N 5∘191015840294010158401015840 and E103∘081015840272910158401015840) from Peninsular Malaysia The temperaturewas 28ndash33∘C with pH between 560 and 820 salinity 0 pptdissolved oxygen in the range of 200ndash1100mgL turbidityin the range of 100ndash1800NTU and conductivity 054-055mScmThe fishes were verified by Terengganu FisheriesDepartment Ministry of Agriculture Malaysia Fillets wereobtained by carefully cutting the fish lengthwise along back-bone to gain maximum amount of flesh The whole fish filletextract was prepared by using pressure cooker set at 100∘Cfor 30min The fresh boneless fillet was weighed and placedon a stainless steel wire mesh mounted on a stainless steeltripod in the pressure cooker Fish fillet and distilled waterwere added in volume ratio of 1 2 The extract was obtainedthrough steaming At the end of the extraction procedurethe fillet was discarded while collecting the liquid extractusing Whatman no 1 filter paper The obtained extract wasfreeze dried using a freeze drier and stored at 20∘C until use[7 8]

22 Drugs and Chemicals Chloroform AR phenolphthaleinpH indicator solution and sodium hydroxide pellets wereprocured from SD Fine-Chem Limited Mumbai while pureaspirin was obtained from Divis Laboratories HyderabadRanitidine (as Rantac 150mg) was from JB Chemicals andPharmaceuticals Mumbai while surgical spirit was obtainedfrom Kakatiya Pharma Hyderabad Topferrsquos reagent anddistilled water were obtained from Nice Chemicals Chinaand Stangen Fine Chemicals Hyderabad respectively Allchemicals were used without further purification

23 Animals For the evaluation of the antiulcer propertymale albino Wistar rats weighing 160ndash200 g were usedThe study was conducted after the approval of protocol byInstitutional Animal Ethical Committee (IAEC)The animalswere maintained under standard conditions that is housedin polypropylene cages and maintained at a temperature27 plusmn 2

∘C relative humidity 65 plusmn 10 under 12-hour lightand dark cycle The animals were fed with standard pelletdiet with water ad libitum in an animal house (Reg num-ber 1534POa11CPCSEA) approved by the Committee for

the Purpose of Control and Supervision on ExperimentalAnimals (CPCSEA) The animals were acclimatized for tendays under laboratory conditions before carrying out theexperiments

24 Acute Toxicity and Selection of Test Doses Earlier investi-gations indicate that Channa striata (Bloch) aqueous extractis safe and nontoxic even at the dose of 8 gkg or 100 wvconcentration [7 8] Doses of 30 40 and 50 wv wereselected for the screening of antiulcer activity based on thereport of Saleem et al [10]

25 Evaluation of Antiulcer Activity

251 Aspirin Induced Ulcerogenesis in Pylorus Ligated RatsAspirin induced ulcerogenesis in pylorus ligated rats modelwas used for the evaluation of antiulcer activity with slightmodification The animals were divided into five groups (119899 =6) Group I served as negative control and received only vehi-cle Groups II III and IV received aqueous extract ofChannastriata (AECS) at 30 40 and 50 wv concentrationsrespectively orally at the volume of 10mLkgGroupV servedas standard and was treated with standard drug (Ranitidine50mgkg po) [20] Aspirin suspended in 1 CMC in waterwas administered orally at a dose of 500mgkg in 12-hourfasted rats [21] The test extract and standard drug treatmentwere done 30min prior to the administration of aspirin After30min the pyloric ligation surgery was performed Fourhours later the animals were sacrificed by euthanasia

Collection and Measurement of Gastric Juice The stomachswere excised carefully keeping the esophagus closed Thestomachs were opened along the greater curvature removingthe luminal contents The gastric contents were collected andcentrifuged at 1000 rpm for 10 minutes After centrifugationsamples were decanted and the volume of gastric juice wasnoted and is expressed asmL100 g bodyweightThe contentswere subjected to analysis for free and total acidities

Determination of Gastric Juice pH One mL of supernatantliquid was diluted to 10mL with distilled water The pH ofthe solution was recorded with the help of digital pH meter

Estimation of Total and Free Acidities The above solutionwas titrated against 001N NaOH using Topferrsquos reagent asindicatorThe endpoint of the titration was when the solutionturns orange in colour The volume of NaOH was notedwhich corresponds to the free acidity Further the titrationwas continued till the solution regained pink colour Thetotal volume of NaOH was noted which corresponds to totalacidity

Determination of Ulcer Index Mean ulcer score for eachanimal is expressed as ulcer indexThe stomachswerewashedwith running water to see the ulcers in the glandular portionof the stomachThe number of ulcers per stomach was notedand the severity of the ulcers was scoredmicroscopically with

ISRN Pharmacology 3











2O2











3 Results




4 ISRN Pharmacology





526 plusmn 016ns




333 plusmn 033ns


ns



ns3933 plusmn 297

lowast



ns8667 plusmn 120

lowast






lowastlowast

0041 plusmn 00079ns0066 plusmn 00064

lowastlowast





















4 Discussion



ISRN Pharmacology 5

(a) (b) (c) (d) (e)









5 Conclusion


6 ISRN Pharmacology

8

7

6

5

4

3

2

1

0

nsVo

lum

e of g

astr

ic ju

ice (

mL)



(a)

6

4

2

0

ns ns ns

lowastlowast


pH ra

nge

(b)

60

40

20

0

nsns

Volu

me o

f NaO

H (m

L) re

quire

dfo

r neu

tral

izin

g ga

stric

HCl

lowast lowast


(c)



Volu

me o

f NaO

H (m

L) re

quire

dfo

r neu

tral

izin

g ga

stric

HCl

100

80

60

40

20

0

(d)

8

6

4

2

0

lowastlowast


lowastlowast

Num

ber o

f gas

tric

ulc

ers


(e)

ns

lowastlowast

lowastlowast

lowastlowastlowast

010

008

006

004

002

000


Leve

l of c

atal

ase (

units

mg

of ti

ssue

)

(f)

ns

ns

lowast

lowastlowast

010

008

006

004

002

000

Leve

l of s

uper

oxid

e dism

utas

e(u

nits

mg

of ti

ssue

)


AECS 30wvAECS 40wv


(g)

025

020

015

010

005

000


Leve

l of m

alon

dial

dehy

de (n

umbe

r of

mol

es o

f MD

Am

g of

tiss

ue)

AECS 30wvAECS 40wv



(h)


ISRN Pharmacology 7




Acknowledgment


References



























8 ISRN Pharmacology

























Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

ISRN Pharmacology 3











2O2











3 Results




4 ISRN Pharmacology





526 plusmn 016ns




333 plusmn 033ns


ns



ns3933 plusmn 297

lowast



ns8667 plusmn 120

lowast






lowastlowast

0041 plusmn 00079ns0066 plusmn 00064

lowastlowast





















4 Discussion



ISRN Pharmacology 5

(a) (b) (c) (d) (e)









5 Conclusion


6 ISRN Pharmacology

8

7

6

5

4

3

2

1

0

nsVo

lum

e of g

astr

ic ju

ice (

mL)



(a)

6

4

2

0

ns ns ns

lowastlowast


pH ra

nge

(b)

60

40

20

0

nsns

Volu

me o

f NaO

H (m

L) re

quire

dfo

r neu

tral

izin

g ga

stric

HCl

lowast lowast


(c)



Volu

me o

f NaO

H (m

L) re

quire

dfo

r neu

tral

izin

g ga

stric

HCl

100

80

60

40

20

0

(d)

8

6

4

2

0

lowastlowast


lowastlowast

Num

ber o

f gas

tric

ulc

ers


(e)

ns

lowastlowast

lowastlowast

lowastlowastlowast

010

008

006

004

002

000


Leve

l of c

atal

ase (

units

mg

of ti

ssue

)

(f)

ns

ns

lowast

lowastlowast

010

008

006

004

002

000

Leve

l of s

uper

oxid

e dism

utas

e(u

nits

mg

of ti

ssue

)


AECS 30wvAECS 40wv


(g)

025

020

015

010

005

000


Leve

l of m

alon

dial

dehy

de (n

umbe

r of

mol

es o

f MD

Am

g of

tiss

ue)

AECS 30wvAECS 40wv



(h)


ISRN Pharmacology 7




Acknowledgment


References



























8 ISRN Pharmacology

























Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

4 ISRN Pharmacology





526 plusmn 016ns




333 plusmn 033ns


ns



ns3933 plusmn 297

lowast



ns8667 plusmn 120

lowast






lowastlowast

0041 plusmn 00079ns0066 plusmn 00064

lowastlowast





















4 Discussion



ISRN Pharmacology 5

(a) (b) (c) (d) (e)









5 Conclusion


6 ISRN Pharmacology

8

7

6

5

4

3

2

1

0

nsVo

lum

e of g

astr

ic ju

ice (

mL)



(a)

6

4

2

0

ns ns ns

lowastlowast


pH ra

nge

(b)

60

40

20

0

nsns

Volu

me o

f NaO

H (m

L) re

quire

dfo

r neu

tral

izin

g ga

stric

HCl

lowast lowast


(c)



Volu

me o

f NaO

H (m

L) re

quire

dfo

r neu

tral

izin

g ga

stric

HCl

100

80

60

40

20

0

(d)

8

6

4

2

0

lowastlowast


lowastlowast

Num

ber o

f gas

tric

ulc

ers


(e)

ns

lowastlowast

lowastlowast

lowastlowastlowast

010

008

006

004

002

000


Leve

l of c

atal

ase (

units

mg

of ti

ssue

)

(f)

ns

ns

lowast

lowastlowast

010

008

006

004

002

000

Leve

l of s

uper

oxid

e dism

utas

e(u

nits

mg

of ti

ssue

)


AECS 30wvAECS 40wv


(g)

025

020

015

010

005

000


Leve

l of m

alon

dial

dehy

de (n

umbe

r of

mol

es o

f MD

Am

g of

tiss

ue)

AECS 30wvAECS 40wv



(h)


ISRN Pharmacology 7




Acknowledgment


References



























8 ISRN Pharmacology

























Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

ISRN Pharmacology 5

(a) (b) (c) (d) (e)









5 Conclusion


6 ISRN Pharmacology

8

7

6

5

4

3

2

1

0

nsVo

lum

e of g

astr

ic ju

ice (

mL)



(a)

6

4

2

0

ns ns ns

lowastlowast


pH ra

nge

(b)

60

40

20

0

nsns

Volu

me o

f NaO

H (m

L) re

quire

dfo

r neu

tral

izin

g ga

stric

HCl

lowast lowast


(c)



Volu

me o

f NaO

H (m

L) re

quire

dfo

r neu

tral

izin

g ga

stric

HCl

100

80

60

40

20

0

(d)

8

6

4

2

0

lowastlowast


lowastlowast

Num

ber o

f gas

tric

ulc

ers


(e)

ns

lowastlowast

lowastlowast

lowastlowastlowast

010

008

006

004

002

000


Leve

l of c

atal

ase (

units

mg

of ti

ssue

)

(f)

ns

ns

lowast

lowastlowast

010

008

006

004

002

000

Leve

l of s

uper

oxid

e dism

utas

e(u

nits

mg

of ti

ssue

)


AECS 30wvAECS 40wv


(g)

025

020

015

010

005

000


Leve

l of m

alon

dial

dehy

de (n

umbe

r of

mol

es o

f MD

Am

g of

tiss

ue)

AECS 30wvAECS 40wv



(h)


ISRN Pharmacology 7




Acknowledgment


References



























8 ISRN Pharmacology

























Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

6 ISRN Pharmacology

8

7

6

5

4

3

2

1

0

nsVo

lum

e of g

astr

ic ju

ice (

mL)



(a)

6

4

2

0

ns ns ns

lowastlowast


pH ra

nge

(b)

60

40

20

0

nsns

Volu

me o

f NaO

H (m

L) re

quire

dfo

r neu

tral

izin

g ga

stric

HCl

lowast lowast


(c)



Volu

me o

f NaO

H (m

L) re

quire

dfo

r neu

tral

izin

g ga

stric

HCl

100

80

60

40

20

0

(d)

8

6

4

2

0

lowastlowast


lowastlowast

Num

ber o

f gas

tric

ulc

ers


(e)

ns

lowastlowast

lowastlowast

lowastlowastlowast

010

008

006

004

002

000


Leve

l of c

atal

ase (

units

mg

of ti

ssue

)

(f)

ns

ns

lowast

lowastlowast

010

008

006

004

002

000

Leve

l of s

uper

oxid

e dism

utas

e(u

nits

mg

of ti

ssue

)


AECS 30wvAECS 40wv


(g)

025

020

015

010

005

000


Leve

l of m

alon

dial

dehy

de (n

umbe

r of

mol

es o

f MD

Am

g of

tiss

ue)

AECS 30wvAECS 40wv



(h)


ISRN Pharmacology 7




Acknowledgment


References



























8 ISRN Pharmacology

























Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

ISRN Pharmacology 7




Acknowledgment


References



























8 ISRN Pharmacology

























Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

8 ISRN Pharmacology

























Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of





Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

research article gastroprotective effect of freeze dried stripped...

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