金匮肾气丸对肾阳虚证雄性大鼠生精细胞中凋亡相关蛋白Bcl-2、Bax的影响

Effect of Jingguishenqi pill on expression of apoptosis-related protein Bcl-2 and Bax in male mouse with kidney yang deficiency

背景:肾虚可导致不育,金匮肾气丸是治疗肾阳虚证的经典药物,但其作用机制尚不明确。目的:观察金匮肾气丸对凋亡相关蛋白Bcl-2、Bax在肾阳虚证雄性大鼠生精细胞中表达的影响。方法:成年雄性SD大鼠100只随机分成5组,采用氢化可的松注射液制备肾阳虚模型。金匮肾气丸低、中、高剂量组在造模基础上每天灌胃含生药0.625,1.250,2.500g/kg的肾气丸混悬液,连续30d。正常组和模型组灌胃等量蒸馏水。检测各组大鼠左侧睾丸质量、血清睾丸酮水平、精子密度、精子活率改变情况。采用免疫组织化学方法检测Bcl-2、Bax蛋白在生精细胞中的表达。结果与结论:模型组左侧睾丸质量、血清睾丸酮水平、精子密度、精子活率低于正常组、金匮肾气丸中、高剂量组(P<0.05)。与模型组比较,正常组、金匮肾气丸低、中、高剂量组大鼠生精细胞中Bcl-2蛋白表达显著增高(P<0.05),Bax蛋白表达显著降低(P<0.05)。说明金匮肾气丸可使肾阳虚证雄性大鼠生精细胞中Bcl-2蛋白表达升高,Bax蛋白表达降低,从而抑制细胞凋亡,对肾阳虚证雄性大鼠生殖能力有较好的治疗作用。

BACKGROUND： Kidney-deficiency causes sterility. Jingguishenqi pill is a classical Chinese traditional medicine for treating kidney yang deficiency, but its mechanism remains poorly understood. OBJECTIVE： To explore the effect of Jingguishenqi pill on expression of Bcl-2 and Bax in male mice permatogenic cells with kidney yang deficiency. METHODS： Totally 100 mature male rats were divided into 5 groups randomly： blank control group （normal group）, model group, Jingguishenqi pill with low dosage group, middle dosage group, high dosage group. Except the normal group, the others were made into model of kidney-yang deficiency by injecting hydrocortisone. After the model preparation, the mice in the normal and model groups were fed with distilled waters, and those in the Jingguishenqi pill with different dosage groups were fed with the same dose of medicine （0.625, 1.250 and 2.500g/kg） for 30 days continuously. Then, the weight of the left testis was measured, and the blood level of testosterone, and the sperm density, survival rate were examined. Then the expression of Bcl-2 and Bax in spermatogenic cells were investigated with the immunohistochemistry and image analysis. RESULTS AND CONCLUSION： The weight of left testis, blood level of testosterone, sperm density, sperm survival rate in the model group were decreased than that in the control, Jingguishenqi pill with low dosage group, middle dosage group, high dosage group （P〈0.05）. The number of cells of expressing Bcl-2 and the intensity of its expression in the model group were lower than that in other four groups （P〈0.05）. The number of cells of expressing Bax and the intensity of its expression in the model group were higher than that in other four groups （P〈0.05）. Jingguishenqi pill can promote the expression of Bcl-2 but reduce Bax expression, which exerts a certain therapeutic effect on the kidney-yang deficiency in rats.