摘要
目的研究肾气丸对衰老所致雄性大鼠睾丸支持细胞紧密连接功能损伤的保护作用及其可能的分子机制。方法将SPF级SD雄性大鼠随机分为4组,分别为青年对照组(2月龄)、衰老模型组(17月龄)及肾气丸低、高剂量组(0.1、0.4 g·kg^(-1),17月龄),每组8只,分别给予普通饲料或含药饲料持续喂养4个月。HE染色法观察睾丸组织形态变化并测量生精上皮厚度和生精小管直径;透射电镜观察睾丸组织紧密连接超微结构;Western Blot法检测睾丸组织中波形蛋白(Vimentin)、闭合蛋白(Occludin)、闭锁连接蛋白-1(Zonula occludens-1,ZO-1)和β-连环蛋白(β-catenin)表达水平;双标免疫荧光法检测睾丸支持细胞中Vimentin、β-catenin、 ZO-1和磷酸化p38丝裂原活化蛋白激酶(Phosphorylated p38 Mitogen-activated protein kinase,p-p38MAPK)的表达和定位。结果 HE结果显示,肾气丸能明显改善自然衰老大鼠睾丸组织形态学变化,提高生精上皮厚度(P<0.05)和生精小管直径(P<0.05,P<0.01);透射电镜结果显示,肾气丸能明显改善自然衰老大鼠支持细胞紧密连接超微结构;Western Blot结果显示,肾气丸能明显上调自然衰老大鼠睾丸组织中Vimentin、Occludin、ZO-1和β-catenin蛋白表达水平(P<0.05,P<0.01,P<0.001);免疫荧光结果显示,肾气丸能明显上调衰老大鼠睾丸支持细胞Vimentin、ZO-1和β-catenin蛋白表达水平,下调支持细胞p-p38MAPK蛋白表达水平。结论肾气丸可减轻自然衰老大鼠睾丸支持细胞紧密连接损伤,其机制可能与下调支持细胞p38MAPK信号通路有关。
Objective To investigate the protective effects on Shenqi pills against injury of tight junction function of sertoli cells in the testis of aging rats and explore the possible mechanism.Methods The male SD rats were randomly divided into 4 groups,with 8 rats in each group:adult control group(2 months old),aging model group(17 months old),low and high dose of Shenqi pills-treated groups(0.1 g·kg^(-1),0.4 g·kg^(-1),17 months old). Rats in the adult control group and the aging model group were fed with normal diet for 4 months. Simultaneously,rats in Shenqi pills-treated groups were given medicated feed for 4 months. The morphological changes of testis were observed by HE staining, the thickness of seminiferous epithelium and the diameter of seminiferous tubules were detected. Tight junction ultrastructure of Sertoli cells changes were observed by transmission electron microscope. In addition,the expression levels of Vimentin,Occludin,Zonula occludens-1(ZO-1)and β-catenin in testis were detected by Western Blot. The expression and localization of Vimentin,ZO-1,β-catenin and phosphorylated p38 Mitogen-activated protein kinase(p-p38 MAPK)in testis were detected by double-labeling immunofluorescence.Results HE staining results showed that Shenqi pills significantly improved the structure of testicular tissue of aging rats, increased the thickness of seminiferous epithelium(P<0.05) and the diameter of seminiferous tubule(P<0.05,P<0.01). Transmission electron microscopy results showed that Shenqi pills also improved the ultrastructure of testicular sertoli cells tight junction in aging rats. In addition, Western Blot results showed that Shenqi pills significantly increased the relative protein expression levels of Vimentin,Occludin,ZO-1 and β-catenin(P<0.05,P<0.01, P<0.001)in testis. Moerover, immunofluorescence results showed that Shenqi pills remarkably upregulated the protein expression levels of Vimentin,ZO-1 and β-catenin in Sertoli cells and downregulated the protein expression of p-p38 MAPK in sertoli cells
作者
张艳
张长城
杨圆
吴杰
袁丁
赵海霞
ZHANG Yan;ZHANG Changcheng;YANG Yuan;WU Jie;YUAN Ding;ZHAO Haixia(Medical College of China Three Gorges University,Yichang 443002 Hubei,China;Third Level Laboratory of Chinese Medicine Pharmacology(tumor)Research,State Administration of Traditional Chinese Medicine,China Three Gorges University,Yichang 443002 Hubei,China;Analysis and Testing Center of China Three Gorges University,Yichang 443002 Hubei,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2021年第6期783-790,共8页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金项目(81774316,81873077)。
关键词
肾气丸
衰老
睾丸
支持细胞
紧密连接
大鼠
Shenqi pills
aging
testis
sertoli cells
tight junction
rats
