基于内质网应激研究五子衍宗方对自然衰老大鼠睾丸生殖细胞凋亡的保护作用  

Protective effect of Wuzi Yanzong recipe on testicular germ cell apoptosis in natural ageing rats through endoplasmic reticulum stress

作　　者：马娜 张长城[1] 陈茜[1] 马琼艳 尤旭 杨思琪 袁丁[2] 赵海霞[1] MA Na;ZHANG Chang-cheng;CHEN Qian;MA Qiong-yan;YOU Xu;YANG Si-qi;YUAN Ding;ZHAO Hai-xia(Medical College of China Three Gorges University,Yichang 443002,China;Ren-He Hospital of China Three Gorges University,Yichang 443002,China)

机构地区：[1]三峡大学医学院,湖北宜昌443002 [2]三峡大学仁和医院,湖北宜昌443002

出　　处：《中国中药杂志》2018年第19期3899-3904,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81573931,81373881,81774316,81503334);三峡大学学位论文培优基金项目(2018SSPY095)

摘　　要：基于内质网应激(ERS)研究五子衍宗方对自然衰老大鼠睾丸生殖细胞凋亡的保护作用。将16月龄SPF级SD雄性大鼠随机分为3组,自然衰老组(Ageing),五子衍宗方低剂量组(WZ,1 g·kg^-1)和五子衍宗方高剂量组(WZ,4 g·kg^-1),另购买2月龄SD雄性大鼠作为青年对照组(Adult),每组10只。给予普通饲料或含药饲料喂养4个月后,处死大鼠,取出睾丸组织备用。苏木素-伊红(HE)染色观察睾丸组织形态;Western blot法检测睾丸组织中内质网应激相关蛋白GRP78,p-PERK,p-eif2α,ATF4,p-IRE1,XBP1,ATF6及其介导的凋亡相关蛋白CHOP,caspase 12,p-JNK的表达水平。HE结果显示,五子衍宗方能明显改善自然衰老所致的睾丸组织形态变化。Western blot结果显示,与自然衰老组相比,五子衍宗方能显著升高自然衰老大鼠睾丸组织内质网应激相关蛋白GRP78,p-PERK,p-eif2α,ATF4,p-IRE1,XBP1,ATF6的表达水平;显著降低内质网应激介导的凋亡相关蛋白CHOP,caspase 12,p-JNK的表达水平。综上所述,五子衍宗方可通过调节内质网应激减轻自然衰老大鼠睾丸生殖细胞凋亡。To study the protective effects of Wuzi Yanzong recipe on testis germ cell apoptosis in natural ageing rats through endoplasmic reticulum stress(ERS), 16-month-old male SPF grade SD rats were randomly divided into three groups: ageing model group, and low and high-dose Wuzi Yanzong recipe groups(WZ, 1 and 4 g·kg^-1), with 10 rats in each group. In addition, 2-month-old SD male rats were used as adult control group. The ageing model group and the adult control group were fed with normal diet for 4 months. WZ groups were given the medicated feed for 4 months. After fasting for 12 hours, the rats were put to death. Then, the testes were immediately collected. The change of testicular tissue morphology was observed by HE staining. The expression levels of ER stress-related proteins GRP78, p-PERK, p-eif2α, ATF4, p-IRE1, XBP1, ATF6 and apoptosis-related proteins CHOP, caspase12 and p-JNK in testes were detected by Western blot. Compared with the ageing model group, Wuzi Yanzong recipe alleviated the morphological changes of testicular tissue. Western blot results showed that Wuzi Yanzong recipe significantly increased the expression levels of endoplasmic reticulum stress-related proteins GRP78, p-PERK, p-eif2α, ATF4, p-IRE1, XBP1, ATF6 and significantly decreased the expression levels of endoplasmic reticulum-induced apoptosis-related proteins CHOP, caspase 12 and p-JNK. In conclusion, Wuzi Yanzong recipe can alleviate the ageing-related apoptosis of testicular germ cells in natural ageing rats by regulating endoplasmic reticulum stress.

关 键 词：五子衍宗方 内质网应激 衰老 睾丸 凋亡 

分 类 号：R285.5[医药卫生—中药学]