the protective effect of ginger and n- acetyl cysteine on ciprofloxacin-induced reproductive...
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The Protective effect of Ginger and N- Acetyl
Cysteine on Ciprofloxacin-Induced Reproductive
Toxicityin Male Rats
Nashwa, A. Abu-Aita1, Kawkab, A. Ahmed and
Samar M Mouneir
IntroductionIntroduction
Infertility is one of the major health problems in animals. Several conditions can interfere with spermatogenesis and reduce sperm quality and production. Drug treatment, chemotherapy, toxins and environmental factors can have harmful effect on spermatogenesis and sperm normal production Antibiotics are used in the treatment of many infections and been shown to significantly affect semen parameters in human and animal models
Ciprofloxacin (CPX) is one of the newer generations of antibacterial agents. It is used to combat various infectious diseases in man and animals.Recently, it has been observed that the oxidative damage to testicular cells induced by variousxenobiotics can result in testicular dysfunction and male infertility.
Ginger is a strong anti-oxidant substance and may either mitigate or prevent generation of free radicals. It is considered a safe herbal medicine with only insignificant adverse side effects. It have immuno-modulatory, anti-tumorigenic, Anti-inflammatory, anti-apoptotic, anti-hyperglycemic antilipidemic and anti-emetic actions.
N-acetyle cysteine (NAC), a potent antioxidant Derives from the amino acid L-Cysteine. It has been used clinically for decades for the treatment of manydiseases. It has been also used as a chelator of heavy metals to protect against oxidative stress and prevent damage of cells.
Aim of workAim of work
to evaluate the reproductive toxicity induced by ciprofloxacin antibiotic and the protective effect of ginger and/or N acetyl cysteine in male rats.
Materials and MethodsMaterials and MethodsExperimental groups:49 Rats (120-150g) were randomly divided into (7) groupsGroup(1): kept as a control.Group(2): was daily administered ginger at a dose of 100 mg⁄kg.b.wt for 65 days (5 days/week).Group(3) was daily administered N-acetyl cysteine (NAC) at a dose of 50 mg⁄kg.b.wt Group(4) was orally administered ciprofloxacin (CPX) at a dose of 12.5 mg⁄kg.b.wt. Group(5) was concurrently administered ciprofloxacin(12.5 mg⁄kg.b.wt) with ginger at a dose of 100 mg⁄kg.b.wt. Group(6) was concurrently administered ciprofloxacin(12.5mg⁄kg.b.wt) with N-acetyl cysteine (50mg⁄kg.b.wt) Group(7) was concurrently administered ciprofloxacin(12.5mg⁄kg.b.wt.) with ginger (100mg⁄kg.b.wt.) and N acetyl cysteine (50 mg⁄kg.b.wt)
Body weight:At the end of the experimental period, the body weight of each
individual rat was measured.
Reproductive organ indices: Reproductive organs weight:Rats were sacrificed after the last day of treatment and reproductive
organs; testes, epididymis and seminal vesicles were weighed.Sperm functions analysis:Semen samples were collected from cauda epididymis and used for
sperm functions analysis.
Blood samples:Blood samples were collected from the retroorbital venous plexus
of rats. Centrifuged at 3000 rpm for 10minutes for serum separation. Serum was used for biochemical and hormonal (Testosterone, Follicle stimulating hormone (FSH) and Luteinizing
hormone (LH)) studies.
Tissue specimens:Testes were taken from the sacrificed rats inall experimental groups. They were perfused with PBS (phosphate buffered saline), collected in clean dry plastic bags and kept at -40о c for determination of lipid peroxidation (TBARS), reduced glutathione (GSH) and DNA fragmentation test. Specimens from testes,epididymis and seminal vesicles were fixed in 10 % neutral buffered formalin for histopathological study.
Results
Parameter
sControl Ginger NAC CFX
CFX
+Gin
CFX +
NAC
CFX+Gin+
Nac
Body Wt.
(g)
267.0 ±
2.65ab
269.3 ±
3.06a
264.0 ± 3.61b
268.7
±2.31ab
270.3
±4.51a
268.0 ±
2.00ab
269.0 ±
1.00ab
Testes (g) 1.61± 0.03b
1.72±
0.07a
1.59+0.04
b
1.26±
0.05e
1.47±
0.03c
1.40±
0.01d
1.65± 0.04ab
Epididymis
(g)
0.36±
0.04ab
0.41±
0.03a
0.35±
0.04ab
0.22±
0.03c
0.32±
0.03b
0.31±
0.05b
0.37± 0.03ab
Seminal
ves.(g)0.60± 0.04a
0.61±
0.06a
0.58±
0.04ab
0.26±
0.03d
0.51±
0.03b
0.42±
0.03c
0.54± 0.04ab
Table 1: Effects of ciprofloxacin (CPX), Ginger (Gin.) and N-acetyl cysteine (NAC) on the rat'body weight and weights of reproductive organs of rats.
Means with different superscripts (a-d) within a row are significantly different at P< 0.05LSD least significant difference at probability P< 0.05
Parameters Control Ginger NAC CFXCFX
+Gin
CFX +
NAC
CFX+Gin+N
ac
Sperm count(×106 /ml )
71.36±4.
51a
70.40±4
.51a
65.40±3.
97a
51.6±4.4
5d
67.4±4.67a
b
61.40±3.9
7c
66.4±5.94ab
Motility (%)81.67±2.
68a
79.80±2
.68a
78.80±2.
12a
65.00±3.
61c
79.6±1.52a
75.00±2.1
2b
79.4±4.45a
Viability (%)83.2±2.3
9a
84.00±2
.65a
78.8±3.4
2bc
48.40±3.
05d
79.60±1.1
4bc
77.60±1.8
2c
81.60±2.70ab
Means with different superscripts (a-d) within a row are significantly different at P< 0.05LSD least significant difference at probability P< 0.05
Table 2: Effects of ciprofloxacin (CPX), Ginger (Gin.) and N-acetyl cysteine (NAC) on sperm parameters
Parameters Control Ginger NAC CFXCFX
+Gin
CFX +
NAC
CFX+Gin+
Nc
ACP (U/L)12.67±2.5
7c
15.4±2.6
0bc
14.13±2.2
7bc
22.93±2.
53a
18.37±2.1
8ab
17.7±2.61
b
16.1±3.57bc
LDH (U/L)208.90 ±
1.87e
206.13±1
.80e
205.47 ±
2.84e
300.8 ±
2.88a
259.13 ±
2.72b
223.33 ±
3.06c
214.93 ±
2.61 d
FSH
(mIu/ml)
0.65+0.04
a
0.62+0.0
4 ab
0.58+0.05b
c
0.36+0.0
4e
0.62+0.04a
b
0.48+0.03
d
0.55+0.03c
LH
(mIu/ml)
0.19 ±
0.04 a
0.20 ±
0.02 a
0.14 ±
0.03 bc
0.05 ±
0.02 d
0.16 ±
0.02 ab
0.10 ±
0.02 c
0.11 ± 0.02
c
Testostero
ne
(ng/ml)
1.89±0.11a
1.79±0.1
7ab
1.48±0.15c
d
0.99±0.1
0e
1.75±0.08a
bc
1.58±0.17
bcd
1.35±0.16d
Table 3: Effects of ciprofloxacin (CPX), Ginger (Gin.) and N-acetyl cysteine (NAC) on some serum enzymes and FSH, LH and Testosterone hormones:
Means with different superscripts (a-e) within a row are significantly different at P< 0.05LSD least significant difference at probability P< 0.05
Parameters Control Ginger NAC CFX CFX +Gin CFX + NACCFX+Gin+
Nac
MDA (mmol / g.
tissue)18.67±3.57e 15.18±3.0e 19.13±3.25de 39.95 ± 3.09a 25.63±3.29bc 30±2.00b 24.33+2.52cd
GSH (mmol / g.
tissue)0.28±0.04ab 0.32±0.04a 0.23±0.04bc 0.08±0.02e 0.19±0.03cd 0.15±0.04d 0.21+0.04cd
DNA FRG.
(%)48.00±0.83d 47.78±1.6d 45.56±0.93e 64.74±1.51a 53.98±0.90c 59.36±1.19b 52.44+1.16c
Table 4: Effects of ciprofloxacin (CPX), Ginger (Gin.) and N-acetyl cysteine (NAC) on testicular Malondialdhyde (MDA), reduced glutathione (GSH) and DNA fragmentations:
Means with different superscripts (a-e) within a row are significantly different at P< 0.05LSD least significant difference at probability P< 0.05
The DNA was electrophoresed on TAE agarose gel for one hour at 80 V. The DNA fragmentation pattern was visualized by ethidium bromide. Lane1-2: CPX, Lane3: DNA ladder, Lane4-5: Control, Lane6-7: Ginger,Lane8-9: NAC and Lane10-11: CPX+Gin.+NAC
Histopathological results
ConclusionConclusion
CPX induces reproductive toxicity in rats Through Generating oxidative damage. It induces an adverse effect on reproductiveorgans weights,sperm parameters (sperm count, sperm motility and viability), reproductive hormones (testosterone, LH, and FSH)and histopathological alterations.Ginger and NAC have an important role in ameliorating reproductive toxicity induced by CPX through restoring the oxidant antioxidant balance.
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