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.