五子衍宗方对自然衰老大鼠睾丸生殖细胞DNA氧化损伤的保护作用研究  

Protective effect of Wuzi Yanzong recipe on testicular DNA oxidative damage in natural ageing rats

作　　者：马娜 赵海霞[1] 陈茜[1] 尤旭 马琼艳 杨思琪 袁丁[2] 张长城[1] MA Na;ZHAO Hai-xia;CHEN Qian;YOU Xu;MA Qiong-yan;YANG Si-qi;YUAN Ding;ZHANG Chang-cheng(Medical College,China Three Gorges University,Yichang 443002,China;Ren-He Hospital,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14期2985-2990,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(8150334,81573931,81774316,81373881);三峡大学学位论文培优基金项目(2018SSPY095)

摘　　要：基于Nrf2/HO-1通路和碱基切除修复（BER）研究五子衍宗方对自然衰老大鼠睾丸生殖细胞DNA氧化损伤的保护作用。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）,2月龄SD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饲养4个月后,处死大鼠,取出睾丸组织备用。采用黄嘌呤氧化酶法和硫代巴比妥酸（TBA）法检测睾丸组织中超氧化物歧化酶（SOD）活力和丙二醛（MDA）含量;免疫荧光检测睾丸组织中8-羟基脱氧鸟苷（8-OHd G）和核因子E2相关因子2（Nrf2）的表达和定位;Western blot法检测睾丸组织中Nrf2及其下游相关蛋白血红素氧合酶-1（HO-1）和醌氧化还原酶（NQO1）的表达水平,BER相关蛋白脱嘌呤脱嘧啶核酸内切酶（APE1）、8-羟基鸟嘌呤DNA糖苷酶1（OGG1）和X线修复交错互补基因1（XRCC1）表达的变化情况。结果显示,五子衍宗方能显著提高睾丸SOD活力,降低MDA含量;免疫荧光结果显示,五子衍宗方能明显上调Nrf2的表达,下调8-OHd G的表达;Western blot结果显示,五子衍宗方能显著上调Nrf2,HO-1,NQO1的蛋白表达,显著下调APE1,OGG1,XRCC1的蛋白表达水平。综上所述,五子衍宗方可通过调节Nrf2/HO-1和碱基切除修复通路减轻自然衰老大鼠睾丸DNA氧化损伤。To study the protective effects of Wuzi Yanzong recipe on DNA oxidative damage of testis germ cells in natural ageing rats based on Nrf2/HO-1 signaling pathway and base excision repair（BER）. In the study,16-month-old SPF grade male SD rats were randomly divided into three groups,namely ageing model group,and low and high-dose Wuzi Yanzong recipe groups（WZ,1,4 g·kg^-1）. In addition,2-month-old SD rats were used as adult control group（10 rats in each group）. The ageing model group and the adult control group were fed with normal diet for 4 months. WZ groups were given medicated feed for 4 months. After fasting for 12 hours,the rats were put to death. Then,the testes were immediately removed. The vitality of superoxide dismutase（SOD） and malondialdehyde（MDA） content in testis were detected by xanthine oxidase method and thiobarbituric acid（TBA） method. The levels of Nrf2 and 8-OHd G were detected by immunofluorescence. The protein expression levels of Nrf2,HO-1,NQO1,APE1,OGG1 and XRCC1 were detected by Western blot. Compared with the ageing model group,WZ significantly increased the SOD vitality and decreased MDA content of testis. In addition,immunofluorescence results showed that WZ significantly attenuated testicular DNA oxidative damage and improved antioxidant capacity. Such changes were accompanied by the down-regulation of DNA oxidative damage response protein 8-OHd G levels and the up-regulation of Nrf2 levels. Moreover,Western blot results showed that WZ significantly increased the protein expression levels of Nrf2,HO-1 and NQO1 of the testis germ cells,when compared with ageing model group. In parallel,the protein expression levels of APE1,OGG1 and XRCC1 were significantly decreased. In conclusion,WZ improves ageingrelated DNA oxidative damage via Nrf2/HO-1 and BER pathways.

关 键 词：五子衍宗方 Nrf2/HO-1通路 碱基切除修复(BER) 衰老 睾丸 DNA损伤 

分 类 号：R285.5[医药卫生—中药学]