Effects of Aqueous Extract of Allium Cepa (Onion) on Ciprofloxacin-induced Testicular Toxicity in Wistar Rats

Effects of Aqueous Extract of Allium Cepa (Onion) on Ciprofloxacin-induced Testicular Toxicity in Wistar Rats

CHAPTER ONE

Aim and objectives of the study

Aim of the study

The aim of this study is to investigate the effects of the Allium cepaextract on ciprofloxacin- induced testicular toxicity in Wistar rats.

Objectives of the study

The objectives of the study are to investigate the effects of

1. Allium cepa extracton sperm apoptosis using acridine orange and fluorescence
2. Allium cepa extract on changes in the testis histology and sperm parameters due to ciprofloxacin
3. Allium cepa extract on testicular apoptosis induced by ciprofloxacin using caspase-3 activity
4. Allium cepaextract on oxidative stress
5. Allium cepa extract and ciprofloxacin on haematological

CHAPTER TWO 

LITERATURE REVIEW

Spermatogenesis

Spermatogenesis is the process by which a complex, interdependent population of germ cells produces spermatozoa. Mammalian spermatogenesis can be divided into two phases (Rodriguez et al., 1997). The first phase of spermatogenesis starts after birth. It is characterized by the sequential appearance of cells within the seminiferous tubules, which corresponds to each stage of the germinal cell. The second phase starts during puberty (ongoing phase), characterized by the concomitant presence of stem cells and early spermatogonia to fully differentiated spermatozoa (Rodriguez et al., 1997). Spermatogenesis is a chronological process that takes about 60 days in rats and 72 days in man to complete (Hess, 1990). During this period, the immature germ cellscyclically develop into highly specialized spermatozoa (Rodriguez et al., 1997).

The balance between germ cell proliferation, differentiation and apoptosis is critical to control spermatogenesis. During establishment of spermatogenesis at the puberal age, early germ cells apoptotic wave occurs, aimed at removing abnormal germ cells and maintaining a proper ratio between maturing germ cells and Sertoli cells (Koji, 2001).

Spertmatogonia that are dormant in the seminiferous tubules of the testis since the fetal period begin to increase in number at puberty. After several mitotic divisions, the spermatogonia grow and are transformed into primary spermatocytes. Each primary spermatocyte undergoes the first meiotic division to form two haploid secondary spermatocytes. The secondary spermatocytes undergo a second meiotic division to form four haploid spermatids. The spermatids are gradually transformed into four mature sperms by the process known as spermiogenesis. When spermiogenesisis is complete, the sperms enter the lumina of the seminiferous tubules(Moore and Persaud, 2008).

Sertoli cells lining the seminiferous tubules support and nurture the germ cells and may be involved in the regulation of spermatogenesis. Sperms are transported passively from the seminiferous tubule to the epididymis, where they are stored and become functionally mature. The epididymis is continous with the ductus deferens which transport the sperm to the urethra (Moore and Persaud, 2008).

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CHAPTER THREE

 MATERIALS AND METHODS

Materials 

Experimental animals

Twenty five adult male Wistar rats were obtained from the Department of Pharmacology, Ahmadu Bello University, Zaria. They were kept and maintained under laboratory conditions of temperature, humidity, light and free access to food and water. The animals were allowed to acclimatize for two weeks prior to the commencement of administration of the drug and the extract.

Drugs and reagents

Ciprofloxacin (500 mg/tablet) produced by Fidson Health Care Ltd. was obtained from a reputable pharmaceutical store (Royce Medicare Ltd) in Samaru, Zaria, Kaduna State. Acridine orange and ethidium bromide mixture were bought from Sigma Aldrich Co. UK. Caspase-3 activity kit was ordered from Abcam inc. UK. All other chemicals used were of analytic grade.

CHAPTER FOUR

RESULTS

Morphological Studies

Weights of the body and testes

The line graph in Fig. 4.1 shows that there was a continuous increase in the weekly mean weights in all groups. However, in week 2 through week 3 animals in group E which received a low dose of onion only (100 mg/Kg b.w.) showed a decline in mean weight gain.

Also, results from Table 4.1 showed that all the animals used for this study showed a weight gain at the end of the experiment as indicated by the value of body weight differences. However, administration of 22.86 mg/Kg b.w. ciprofloxacin plus 200 mg/Kg b.w. onion extract (group D)led to a significantly (p<0.05) lower percentage gain in weight compared to the control, and to group C which received a low dose of onion plus ciprofloxacin. Rats given 100mg/Kg b.w. of onion extract only(group E) also showed a significantly (p<0.05) lower percentage in weight gain compared to control.

CHAPTER FIVE

DISCUSSION

The present study which aimed at evaluating the effects of Allium cepaextract on ciprofloxacin-induced testicular toxicity explored the effects of the extract on different morphological and biochemical parameters which included body and testis weights, sperm, anti-oxidant and haematological parameters, caspase-3 activity, DNA status and testicular cyto-architecture.

In this study, it was observed that there was a continuous increase in the weekly mean weights of the animals in all the groups. It was also observed that ciprofloxacin-administered rats had no significant gain in weight compared to the control. This result shows that ciprofloxacin has no negative effect on the food consumption and nutrient utilization. This finding is in tandem with the result reported of Abu -Aita et al. (2011).

Allium cepa-administered rats showed a significant (p<0.05) decrease in percentage weight change. This observation agrees with the work of Sung et al. (2011) and Wang et al. (2012). They reported that onion extract significantly reduced body weight, white adipose tissue weight and adipocyte size of the treated mice compared to high–fat induced control mice. This may be due to the inhibition of fatty acid synthase which in turn suppresses lipid accumulation in the body(Sunget al., 2011).

CHAPTER SIX

SUMMARY, CONCLUSION AND RECOMMENDATION

Summary

The major findings of this present work are as follows:

1. Allium cepaextract at low dose showed ameliorative effect on ciprofloxacin- induced testicular toxicity through the following mechanisms:
   1. Reduction of oxidative stress through increase in anti-oxidants i.e.catalase.
   2. Maintenance of testicular protein
   3. Reduction in the level of testicular apoptosis as measured by caspase – 3
   4. Prevention of DNA fragmentation as measured by acridine orange
2. Allium cepaextract at high dose was found to exacerbate the deleterious effect of ciprofloxacin on the
3. Allium cepaextract at both low and high doses was found to reduce body weight- a property that can be exploited in treating obesity related diseases. This is yet to be

Conclusion

Ciprofloxacin induced caspase-dependent apoptosis in the testis and this was ameliorated by the low dose of Allium cepaextract as the high dose of the extract was found to exacerbate the effect of ciprofloxacin.

Recommendation

Further studies should be carried out in the following areas to unravel some of the observations pointed out in this work:

1. Hormonal assay should be carried out to ascertain the role of onion extract on testosterone level .
2. Immuno-histochemical staining of Sertoli cells using anti-tyrosinated tubulin antibody to ascertain the role of onion extract in stabilizing these
3. Other Flourescent dyes such as aniline blue, chromomycin A3 and propidium iodide should also be employed study to the DNA integrity of the sperm cells following administration of ciprofloxacin and onion extract
4. The mechanism through which onion extract brings about reduction in body weight should also be investigated.

REFERENCES

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• Abu-Aita, N.A., Ahmad, K.A. and Mouneir, S.M. (2011). The protective effect of Ginger and N-Acetyl Cystein on ciprofloxacin induced reproductive toxicity in male rat. Journal of American Science, 7(7):741-752.
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