补肾生精方促进少弱精症大鼠精子发生的作用机制研究

Mechanism of Bushen Shengjing Recipe promoting spermatogenesis in rats with oligoasthenospermia

目的:研究补肾生精方对腺嘌呤诱导的少弱精症大鼠精子发生的作用机制。方法:采用腺嘌呤灌胃诱导SD大鼠制备少弱精症大鼠模型,将48只SD大鼠随机分为空白组、模型组、五子衍宗丸组(1 g/kg)和补肾生精方高、中、低剂量组(2.8 g/kg、1.4 g/kg、0.7 g/kg),给药3周。干预结束后采用放射免疫法测定血清睾酮水平;利用计算机辅助精子分析系统评估精子密度、活动力;苏木素-伊红(HE)染色观察大鼠睾丸组织病理形态学改变;免疫组织化学(IHC)检测大鼠睾丸组织支持细胞中波形蛋白表达水平。结果:与模型组相比,给药组的大鼠睾丸病理损伤有所改善,睾丸组织部分生精小管增厚,细胞排列紧密,部分管腔缩小,生精细胞和精子数量增加。与模型组相比,五子衍宗丸组及补肾生精方各剂量组血清睾酮水平均上调(P<0.05),其中,五子衍宗丸组、补肾生精方高剂量组上升明显(P<0.05),差异有统计学意义。与模型组相比,给药后大鼠精子质量有所改善,精子活动力差异有统计学意义(P<0.05),同时,补肾生精方高剂量组在改善精子密度方面优于模型组(P<0.05);补肾生精方高剂量组在改善精子活动力方面优于五子衍宗丸组,差异有统计学意义(P<0.05)。与模型组相比,给药后各组的睾丸组织支持细胞中波形蛋白表达上调(P<0.05),且补肾生精方高剂量组波形蛋白表达高于五子衍宗丸组(P<0.05)。结论:补肾生精方促进腺嘌呤诱导的少弱精症大鼠精子发生,改善病理损伤与精子质量,其机制与上调支持细胞波形蛋白表达有关。

Objective To study the effect of Bushen Shengjing Recipe on spermatogenesis in SD rats with oligoasthenospermia induced byadenine and to explore its mechanism.Methods A total of 48 SD rats were randomly divided into blank group,model group,Wuzi Yanzong Pill group(1 g/kg)and high,medium and low dose groups of Bushen Shengjing prescription(2.8 g/kg,1.4 g/kg,0.7 g/kg)according to the random number table method.Serum testosterone level was determined by radioimmunoassay after intervention;computer-aided sperm analysis system was used to evaluate sperm density and motility;hematoxylin-eosin(HE)staining was used to observe the histopathological changes of rat testis;immunohistochemistry(IHC)was used to detect the expression of vimentin in rat testis.Results Compared with the model group,the pathological damage caused by oligoasthenozoospermia in each group was improved after drug administration.Some spermatogenic tubules in testis tissue were thickened,cells were closely arranged,some organs were reduced.The number of spermatogenic cells and sperm were increased.Compared with the model group,the serum testosterone levels in the Wuzi Yanzong Pill group and the Bushen Shengjing Recipe groups were all up-regulated(P<0.05).Among them,the increase in the Wuzi Yanzong Pill group and the high-dose Bushen Shengjing Recipe group was significant,and the difference was statistically significant(P<0.05).Compared with the model group,the sperm quality improved after administration,and the difference in sperm motility was statistically significant(P<0.05),meanwhile,Bushen Shengjing prescription high-dose group was better than model group in improving sperm density(P<0.05);Bushen Shengjing prescription high-dose group was better than Wuzi Yanzong Pill group in improving sperm motility,and the difference was statistically significant(P<0.05).Compared with the model group,the expression of vimentin in Sertoli cells of testis in each group was up-regulated after administration(P<0.05),and the expression of vimentin in high-dose Bushen Shengjing Recipe group was higher than that in Wuzi Yanzong Pill group(P<0.05).Conclusion Bushen Shengjing Recipe can promote the spermatogenesis of rats with oligoasthenospermia induced by adenine,improve pathological damage and sperm quality,and its mechanism may be related to the up-regulation of vimentin expression in supporting cells.