Research On The Toxic Effects Of Pesticide Residues From Saposhnikovia Divaricata And Schisandra

Traditional Chinese Medicine is a precious cultural heritage in our country,has the function such as disease prevention,rehabilitation care and widely used in the market.Recently,pesticide residues in traditional Chinese medicine become the focus of attention problems,whether the pesticide residue in produce toxic effects to the organism has aroused the Chinese government attaches great importance to and the vast majority of scientific workers.So the toxicity of pesticide residue in Chinese medicine research becomes especially important.In view of the above background,the early identification of the staff through Saposhnikovia divaricata and Schisandra from the residual OCPs residues containing ?-BHC,p,p'-DDT and HCB three substances.?-BHC,p,p' DDT,HCB in schisandrae residue content are:31.2 ug/kg,63.1 ug/kg,33.2 ug/kg.?-BHC,p,p'-DDT,HCB in the Saposhnikovia divaricata residue content were 42.35 ug/kg,35.83 ug/kg,21.67 ug/kg.The single contents of the above three kinds of pesticides are accord with national pharmacopoeia 2010 version of the minimum residual standard,but as a result of residual pesticide is a mixture of three kinds of material,blend to did not see study whether the toxic effect,about whether the organism toxic harm to do further research,this experiment is residual pesticide extracts of ovarian granulosa cells(GC)and testicular sertoli cells(SC)toxicity to do the research,GC,SC in mice as the research object,establish in vitro cultivation system,study the effect of extract of reproductive toxicity of pesticide residue.The effect of pesticide residue in Schisandra on ovarian granulosa cell toxicity were studied.Mix through the pancreatic enzyme digestion extract ovarian granulosa cells.With inverted microscope and fluorescence microscope observation of living cells form and training status;HE staining and FSHR immunohistochemical method to identify and extract cells purification;Appraisal results show that the extraction and separation of granulosa cells,high purity can reach more than 80%,to provide effective support for later experiment.With determined by MTT method to detect pesticide extract effect on ovarian granulosa cell toxicity,the results showed that extracts of ovarian granulosa cells for 24,48,72h all have different degree of proliferation,compared with negative control group significant difference(p<0.05),and 48h extremely significantly the rate of cell proliferation(p<0.01).Using Western Blot method to compare pesticide extracts of granular cell ERK1/2,p-ERK1/2 protein expression level,the influence of the results found that pesticides extract could significantly promote ERK1/2,p-ERK1/2 protein expression levels,raised the ERK,p-ERK protein expression levels,extremely significant difference(p<0.01).Therefore,come to the conclusion that pesticide extract can promote ovarian granulosa cell proliferation,and influence of ovarian granular cells ERK1/2,p-ERK1/2 protein expression level of ERK signal transduction pathway of cell caused a certain degree of influence,interfere with the downstream signaling cascade amplifier,disturb the normal physiological function,granular cell ovarian organ injury mice,caused the female reproductive toxicity.Experiments of residual pesticide poisonous to the testicular sertoli cells through the Saposhnikovia divaricata effect was studied,with double Trypsin digestion method,separate the sertoli cells were extracted,using different method for identification.Appraisal results show that the extraction and separation of support cells of high purity,can reach more than 80%,provides effective support for later experiment.Testicular sertoli cells determined by MTT,according to the results of pesticide extract to support cells after 24,48,72h,proliferation rate had significant difference(p<0.05),while proliferation rate was the biggest at 48h,compared with negative control group with significant difference(p<0.01).Western Blot,according to the results of SC,that could express ERK 1/2,p-ERK 1/2 protein in vitro,so the method could used to ERK1/2,p-ERK1/2 protein and the function of the sertoli cells toxicity research.Supported by experimental result,extract on testicular cells after 48h,pesticide extract can significantly promote the testicular support cells ERK 1/2,p-ERK 1/2 protein expression and extremely significant difference compared with negative control group(p<0.01).Therefore,the pesticide extract can promote sertoli cell proliferation,and affect sertoli cells ERK1/2,p-ERK1/2 protein expression levels,caused a certain degree of ERK signal transduction pathway and the effect of support the normal physiological function of cells,damage in mouse.