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目的探讨营养性肥胖对青春期雄性大鼠睾丸生殖细胞凋亡的影响。方法健康青春期Wistar雄性大鼠20只,按随机化原则分为两组,每组10只,对照组给予普通饲料喂养,高脂组用高脂、高热量饲料喂养建立营养性肥胖大鼠模型,10周末处死大鼠摘取睾丸。全自动生化分析仪(ACA)检测外周血总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C);光镜观察睾丸组织病理学改变;原位缺口末端标记法(TUNEL)检测睾丸组织凋亡细胞;免疫组化法检测Cyt c、Caspase-3蛋白的分布和表达;RT-PCR法检测睾丸组织Cyt c mRNA和Caspase-3 mRNA的表达。结果高脂组TC、TG、LDL-C和HDL-C较对照组明显升高,差异有显著性(P<0.05);高脂组生殖细胞凋亡指数明显高于对照组(P<0.01),凋亡细胞以精原细胞和精母细胞为主;高脂组Cyt c蛋白和Cyt c mRNA表达明显高于对照组(P<0.01);高脂组Caspase-3蛋白和Caspase-3 mRNA表达明显高于对照组(P<0.01)。结论营养性肥胖诱导大鼠睾丸生殖细胞凋亡增多,其机制可能与Cyt c、Caspase-3蛋白表达升高相关。
Objective To investigate the effect of nutritional obesity on testicular germ cell apoptosis in adolescent male rats. Methods Twenty healthy male Wistar rats were randomly divided into two groups (n = 10 in each group). The control group was fed with normal diet and the high fat diet group was fed with high fat and high calorie diet to establish a model of nutritional obesity. Rats were sacrificed 10 weeks after the removal of testicular. Peripheral blood total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) were detected by automatic biochemical analyzer (ACA) The changes of histopathology were observed. The apoptotic cells in testis were detected by TUNEL. The distribution and expression of Cyt c, Caspase-3 protein were detected by immunohistochemistry. Cyt c mRNA and Caspase -3 mRNA expression. Results The levels of TC, TG, LDL-C and HDL-C in hyperlipidemia group were significantly higher than those in control group (P <0.05). The germ cell apoptosis index in hyperlipidemia group was significantly higher than that in control group (P <0.01) (P <0.01). The expression of Caspase-3 protein and Caspase-3 mRNA in high fat diet group were significantly higher than those in control group Obviously higher than the control group (P <0.01). Conclusion Nutritional obesity induces more germ cell apoptosis in testis, which may be related to the increased expression of Cyt c, Caspase-3 protein.