Ameliorating effect of emblica officinalis on testosterone and sperm toxicity of endosulfan induced mice

Introduction Among pesticides and their related chemicals, organochlorine insecticides have so far drawn the primary attention. Dischlorodiphenyl Trichloroethane (DDT), dieldrin, and toxaphene have instrinsic estrognic activity and are known as possibly endocrine-disrupting chemicals. (Soto et al, 1994). Many attempts have been made to study the toxic effects of pesticides on the reproductive organs of mice (Kumar and Nath 1995, Russel 1995). Endosulfan may cause decrease in semen quality, increase in testicular and prostrate cancer and an increase in defects in male sex organs (Hileman 1994, Solo 1993). Biochemical changes in endosulfan treated testes of rats was observed by (Sinha et al 1995). Endosulfan treatment in pubertal rate inhibits testicular functions (Chitra et al 1999). Endosulfan is an organochlorine pesticide which is widely used by farmers in India for pest control. It has been well reported that endosulfan causes degradation of testicular cells in laboratory animals (Khan & Sinha 1996, Sinha et al 1997 & Sinha et al 2004). Such chemicals have been found to cause reproductive health problems & apoptosis in both humans & wild life (Murray et al 2001). Testosterone level steadily drops in men (Scelto, 2006). To evaluate a comparative ameliorating effect, if any, to mitigate the sperm toxicity and hormonal imbalance in mice for which Emblica officinalis (medicinal plants) have been taken as curative measure : Emblica officinalis leaves and fruit have been used for fever and inflammatory treatments by rural populations in its growing areas. The earlier study have demonstrated potent anti-microbial (Ahmad et al, 1998), antioxidant (Bhattacharya et al, 1999), adaptogenic (Rege et al, 1999), hepatoprotective (Jeena et al, 1999), anti-tumor (Jose et al, 2001) and anti-ulcerogenic activities (Sairam et al, 2002) in the fruits of Emblica officinalis, leaf extracts have been shown to posses anti-inflammatory activity (Asmawi et al, 1993; Jhantola-Vormisto et al, 1997). Material & Methods In the present investigation, experiments were performed on 3-4 months old healthy male Swiss albino mice, Mus musculus. For the optimal growth and reproduction the mice were kept in ideal condition in the laboratory. The oral LD50 value of endosulfan for mice was estimated by standard interpolation method, which was 7 mg / kg b.w. The standard data reference for LD50 of endosulfan for mice is 7.36 mg/ b.w. (EXTOXINET, 1996). Endosulfan manufactured by Excel Industries Mumbai (E.C 35%) was dissolved in distilled water to prepare sublethal dose of 3 mg/kg b.w was administered by gavage method. A vehicle of control group of mice was established and served with equal volume of distilled water by gavage method. The treated and control group were sacrificed on 35 th day of treatment. The sperm count was done through cauda epididymis. Sperm Count The Cauda epididymis was dissected out and washed thoroughly in normal Saline (0.85 %). Cauda epididymis was incised at several places in 1 ml distilled water so as to allow the sperm to ooze out. After that, two drops of Eosin Y is mixed well with sperm. Sperm counts was made using an improved Neubauer’s chamber taking a drop of above preparation in it & observed at 450x magnification. Sperm Motility Cauda epididymis was dissected out and ruptured on microscopic slide. After covering it with a cover slip, the motility of the spermatozoa was examined. Estimation of Testosterone Level In Serum of Mice: By Elisa Technique Blood sample were collected after each sacrifice and there serum were isolated. Using the ELISA method Testosterone kit of LILAC Medicare (P) Ltd., Mumbai was utilized for the experiment.