Research On The Toxic Effect Of Residual Pesticide From Schisandra On Sertoli Cells

In the body inside, environmental estrogen can analog or interference estrogen synthesis, secretion, transport, combination, drain and physiological function, affects the organism physiological effects of estrogen. Environmental estrogen variety, quantity, pollution range is wide, harmfulness, direct endangering the survival of mankind, so environmental estrogen pollution has become a global problem of the pollution of the environment. Although organochlorine pesticides have been banned, but as a result of itself has fat soluble, enrichment sex, degradation difficult, persistent high, can pass the food chain biological enrichment and amplification etc, make the residual organochlorine pesticides in environment, existing for a long time in the human body. DDT, BHC, HCB, three organochlorine pesticides is recognized as the environmental estrogen, has the endocrine interference effect, produce the body estrogen-like effects or antagonistic androgenic effect, enter the body after interference normal physiological function, and further affect the normal physiological process, especially for male reproductive system to produce toxic effect.This topic selection testicular support cell as the research object, more DDT, BHC, HCB, and joint cell toxic effect. Because support cells as testicular features one of the cell, its to the male reproductive system has the vital role, any damage testicular support cells foreign matter will produce a male reproductive toxicity, so, it is male reproductive toxicity of target cells. This experiment by collagenase, pancreatic enzyme double enzyme digestion, extraction mice testis support cells, and HE staining, oil red O staining method and vimentin immunochemical method appraisal separation income cells; Then through the MTT method comparison DDT, BHC, HCB and fruit of Chinese magnoliavine pesticide extract to support cell proliferation, the effects of Using Western blot method to detect DDT, BHC, HCB three kinds of pesticide alone, joint and fruit of Chinese magnoliavine pesticides in extract of male mice testis support cells MAPK signal transduction pathway on related protein ERK1/2, p-ERK1/2, c-Myc protein and Ki67on the distribution of the protein expression.HE staining, oil red O staining method and vimentin immunochemical method appraisal found that the extraction of testicular support cell purity can reach more than80%. MTT results show that DDT, BHC, HCB single and two joint action for testicular support cell value-added role there are no significant difference, DDT, BHC, HCB three mixed and pesticide extract applied to support cells24h,48h,72h, proliferation rate all have significant difference, but in48h proliferation rate when maximum, and the negative control group has very significant difference compared; Therefore, DDT, BHC, HCB three mixed and pesticide extract role in testicular support cells after48h, can have the effect of the largest proliferation. Method of SRB detection:DDT, BHC, HCB alone and combination of the two testicles sertoli cells there are no significant to promote the proliferation of the sertoli cells; DDT, BHC, HCB three mixed and pesticide extract for drug action,24h,48h,72h, proliferation rate all have significant difference, but in48h proliferation rate when maximum, and the negative control group has very significant difference compared; Therefore, DDT, BHC, HCB three mixed and pesticide extract role in testicular support cells after48h, can have the effect of the largest proliferation. The experimental results with the experimental results conform to MTT section.Western Blot method results show that the experimental results show that support cells under the culture condition in vitro can well express ERK1/2, p-ERK1/2protein, visible in vitro experimental method can be used to ERK1/2, p-ERK1/2protein and testicular toxic effect research. By Western Blot experimental result DDT, BHC, HCB joint and extract role in testicular support cells after48h, positive medicine E2, DDT, BHC, HCB joint can significantly improve mice testis support cells ERK1/2, p-ERK1/2protein expression (P<0.05), extract can significantly improve ERK1/2, p-ERK1/2protein expression, and the negative contrast with significant difference (P<0.01).(2)the activation of transcription factors, so that c-Myc protein can also the in vitro culture of conditions good expression, DDT, BHC, HCB joint and extract role in testicular support cells after48h, DDT, BHC, HCB joint can significantly improve mice testis support cells c-Myc protein expression (P<0.05), extract can significantly improve c-Myc protein expression, and the negative contrast with significant difference (P<0.01), and E2group and negative group compared with no difference.(3) proliferation related protein Ki67protein can also the in vitro culture of conditions good expression, DDT, BHC, HCB joint and extract role in testicular support cells after48h, DDT, BHC, HCB joint can significantly improve mice testis support cells Ki67protein expression (P<0.05), extract can significantly improve Ki67protein expression, and the negative contrast with significant difference (P<0.01), and E2group and negative group compared with no difference.