Protein Expression Of Heat Shock Protein 60 In Testis Tissue Of Rat With Diabetic And Hyperglycemia

Objective To establish models of diabetes mellitus and hyperglycemia male Sprayer-Daley rats for studying the micro structure change of seminiferous tubule and the expression of heat shock protein 60,exploring the apotheosis change in testicular sperm cell-sand expression differences of apotheosis associated factors in diabetic and hyperglycemia male Sprayer-Daley rat testis,then explore the possible mechanism.Methods Select 60 Specific pathogen Free(SPF)healthy male Sprayer-Delay rats,each number by computer statistical software after randomly divided into three group:control group(n=15),high-fat group(n=15),diabetes mellitus group(n=30),control group were given standard diet for entire experiment period;high-fat group fed with high-fat food until 168 days,and then add additional 0.2% oxygen pyrimidine into lipid rich food,continue to feed;diabetes mellitus group were given high-fat food before building the model,atthe end of 24 week using streptozocin To establish diabetes model,besides control group and high fat group using sodium cit rate buffer solution instead,diabetes group fed with standard diet after establishing model.Continue to feed until 3 6 week,low density lipoprotein-Cholesterol,Total Cholesterol,Triglyceride,Fasting Blood glucose,high density lipoprotein-Cholesterol and Fasting insulin levels were measured,then deletes unsuccessful rats.The rest of rats were killed after successfully established experimental models,then Separate done side of testicular tissue of rat,put in Bouin's liquid Tissue fixative,after 48 hours embedded biopsy sections,the other side of testicular tissues were put into the frozen tube stored in liquid nitrogen,later move to-80 degrees refrigerator.The expressions of HSP60 proteins in the testis were detected by Strept avidin-biotin-complex(SABC)immunohistochemical staining;observe testicular seminiferous tubule structure change through microscope,the expression level of HSP60 were detected by Western Blot;apotheosis of spermatogenic cells was assessed by means of the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)method?The experimental data is collected and analysis by using SPSS16.0 statistical software,P(all<0.05)means there is significant statistically difference between two experimental data.Results The consequences of Blood glucose monitoring and Blood Lipidremind that both the diabetes mellitus and the hyperthyroidism rats modelwas established successfully.The microscopic view: the sperm cells of control group gave birth to sperm cells arranges from outside to inside according to the principle of development degree,the seminiferous tubule central can be found in a large number of sperm cells.High fat group and diabetes group sperm cells arranged disorderly,a decrease in the number of germ cells,number of sperm cells decreased significantly,some images show there is apart of sperm cells fall off into the tube.Immunohistochemical display HSP60 expression mainly in testicle spermatocyte and head of sperm cell,we did not see it clearly expressed in the cell stroma?compared with control group,the expression of HSP60 was increased significantly in hyperglycemia and diabetes group,high-fat group and diabetes group HSP60 expression quantity compared with no statistical difference;brown of sperm apoplectic cell can be see in each Groups of rats testicles slices,A small amount of positive expression cells scattered distribution in control group rats testicles slices.The rate of testis germ cell apotheosis was increased significantly in hyperglycemia and diabetes group compared with control group?Conclusion The Diabetes and hyperglycemia lead to increase of testicular sperm cells apotheosis and sperm cells reduce.Heat shock protein 60 increased significance in rat testis With hyperglycemia and diabetes.Heat shock protein 60 may be act as a protective protein regulating apotheosis pathway of sperm cells,the pathogenesis of diabetes and hyperglycemia induced testis germ cell apotheosis may be closely related to expression of HSP60 in Germ cell of testistissues.