| Nowadays, environmental endocrine disruptors (EEDs) or endocrine-disrupting chemicals (EDCs) have been paid comprehensive attention to human health, whose harmful effects have become the international hotspot issues to human and wild living beings, also the global research reports have increased rapidly. Pesticide is the one of the big category among EEDs. China, as a big agricultural country, the pesticide output and consumption increases year after year, among numerous kinds of the pesticides, the carbamate and pyrethroid pesticides are widely used, which depend on the advantages that pesticides can rapidly degrade in environment and strong selectivity. However, the proofs reveal that these pesticides can cause dysfunction in reproductive system of human and animals. Therefore, the present study selected carbaryl (Car) and fenvaierate (Fen) as the representatives of carbamateand pyrethroid pesticides, respectively, we discussed the effects of them on steroid hormone and their joint action on Steroidogenesis by in vivo and in vitro studies.Part â… Effects of Carbaryl on Steroidogenesis in OvaryObjectiveTo investigate the effects of Car on the ovarian antioxidant function, steroid hormone production, steroidogenic enzyme mRNA, and protein levels.Methods1. Car was administrated to the adult female Sprague Dawley rats at the doses of 0, 1.028, 5.140 and 25.704 mg·kg-1·d-1 for one month. Vaginal smears of rats were taken to determine estrous cycle. Some organs such as ovary, uterus, brain, heart, liver, spleen and adrenal were dissected and organ coefficients were calculated. Serum 17 β-estradiol (E2) and progesterone (P4) levels were measured by radioimmunoassay. The levels of alanine transaminase (ALT), albumin (ALB), alkaline phosphatase (ALP), aspartate transaminase (AST), cholinesterase (CHE), creatinine (CR), glucose (GLU), Glutathione (GSH), glutathione-s-transferase (GST), total bilirubin (BilT), total cholesterol(TC), total protein (TP) and urea were measured by clinical biochemistry analyzer. The activities of superoxide dismutase (SOD) and glutathione-s-transferase (GST), and the levels of malondialdehyde (MDA) and glutathione (GSH) were measured by spectrophotometry. Total RNA and protein were extracted for RT-PCR and Western blot, respectively.2. Primary cultured human luteinizing-granulosa cells(hGLCs) in vitro were used as model. hGLCs were obtained by follicular aspiration from women taking part in in vitro fertilization programs. The study was approved by the Jiangsu People's Hospital of China. Cells were incubated in different culture plates with Car of various concentrations^, 1, 5, 25 and 125umol/L) for 24h. In some studies, follicle-stimulating hormone (FSH, 200ng/ml) was provided. At the end of all experiments, medium samples were taken and stored at -20 °C for the tests of E2 and P4. And cAMP of cells was abstracted by ethanol. Total RNA was extracted for real-time PCR.Results1. in vivo(1). As compared with the control group, the number of estrous cycle in exposed groups decreased significantly. Proestrus (P) and estrus (E) prolonged but the duration of metestrus (M) and diestrus (D) wasreduced especially after 15 days.(2). In the Car group, the gain of body weight was obviously lower than that of control; meanwhile, the relative organ weights ( ovary , uterus) declined in a dose-dependent manner, but the relative weight of liver enhanced.(3). In the Car group, the levels of ALT, AST, GLU, Urea, ALB, BilT and CHE were changed, especially in 25.704 mg·kg-1·d-1 group.(4). Serum E2 level was lower (especially in 25.704 mg·kg-1·d-1 group) and P4 level was higher (especially in 1.028 mg·kg-1·d-1 group) in exposed groups than that of control group.(5). The activity of SOD reduced both in ovary and serum, the contents of MDA and GSH enhanced in ovary, while reduced in the serum. GST activity enhanced in ovary, however, they first enhanced then reduced in serum.(6). Car could increase the expressions of 3β-hydroxysteroid dehydrogenase (3β-HSD), steroidogenic...
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