二甲双胍调节自噬保护亚急性衰老大鼠睾丸功能的作用  

Effects of metformin on the protection of testicular function by autophagy in subacute aging rats

作　　者：王静 陈冬梅[3] 陈苗[2,4] 虎娜[2,4] 刘文钊 何瑞 崔瑞琴 马会明 WANG Jing;CHEN Dong-mei;CHEN Miao;HU Na;LIU Wen-zhao;HE Rui;CUI Rui-qi;MA Hui-ming(College of Traditional Chinese Medicine,Ningxia Medical University,Yinchuan 750004,China;Key Laboratoryof FertilityPreservationand Maintenance(NXMU),Ministry of Education,Ningxia Medical University,Yinchuan 750004,China;Ningxia Key Laboratory of Stem Cell And Regenerative Medicine,the General Hospital of Ningxia Medical University,Yinchuan750004,China;School of Basic Medical Science,NingxiaMedicalUniversity,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学中医学院,银川750004 [2]宁夏医科大学生育力保持教育部重点实验室,银川750004 [3]宁夏医科大学总医院宁夏干细胞与再生医学重点实验室,银川750004 [4]宁夏医科大学基础医学院,银川750004

出　　处：《中国新药杂志》2022年第17期1752-1760,共9页Chinese Journal of New Drugs

基　　金：宁夏自然科学基金资助项目(2020AAC02016);国家自然科学基金资助项目(81860876);宁夏科技领军人才项目(2020GKLRLX11);宁夏药物研发创新团队项目(NXKJT2019012)。

摘　　要：目的:研究二甲双胍(Met)对D-半乳糖构建的亚急性衰老模型大鼠睾丸功能的影响及其分子调控机制。方法:颈部皮下注射D-半乳糖制备亚急性衰老大鼠模型。次月予以Met溶液灌胃治疗,每周监测大鼠的体重,测定大鼠睾丸重量、睾丸指数;ELISA法检测大鼠血清睾酮(testosterone,T)、卵泡刺激素(FSH)、黄体生成素(LH)和雌二醇(E)水平;HE染色法观察睾丸形态;免疫组化染色检测衰老相关蛋白[β-半乳糖苷酶(β-gal)、p16和p21]、睾丸自噬相关蛋白[磷酸酶张力蛋白同源物诱导激酶1(PINK1)、Parkin、自噬效应蛋白(Beclin-1)、微管相关蛋白1轻链3(LC3)和p62]和睾酮合成关键酶[类固醇激素合成急性调节蛋白(StAR)、细胞色素侧链裂解酶(CYP11A1)和17-β-羟基类固醇脱氢酶3(HSD17β3)]的表达情况,并用Western blot检测上述蛋白的表达量。结果:与对照组相比,衰老模型组和Met组大鼠体重整体呈下降趋势;衰老模型组睾丸重量及指数下降(P<0.01),Met干预后显著上升(P<0.001)。衰老模型组中T,FSH和E水平均显著下降(P<0.05),LH水平明显升高(P<0.01)。模型组衰老相关蛋白β-gal,p16和p21在睾丸细胞质中有明显阳性表达;与衰老模型组相比,Met干预后睾酮合成关键酶StAR,CYP11A1和HSD17β3的表达水平均显著性上升(P<0.05),自噬相关蛋白PINK1,Parkin,Beclin-1和LC3蛋白表达显著升高(P<0.05),p62蛋白表达显著下降(P<0.05)。结论:二甲双胍能激活睾丸自噬水平,增加睾酮合成关键酶的水平,从而预防亚急性衰老大鼠睾丸功能减退。Objective:To investigate the effects of metformin(Met)on testicular function in D-galactose subacute aging rat model and the molecular mechanism of its regulation.Methods:Subacute aging model was established by subcutaneous injection of D-galactose to rats.After modelling for one month,the rats were treated with Met solution by gavage(ig).The body weight of rats was monitored weekly,and testicular weight and testicular index were measured.The serum levels of T,FSH,LH and Ewere measured by ELISA.The morphology of testicular was observed by HE staining.The expression of aging-related proteins(β-galactosidase,p16 and p21),testicular autophagy-related proteins(PINK1,Parkin,Beclin-1,LC3 and p62)and testosterone synthesis-related proteins(StAR,CYP11 A1 and HSD17β3)were detected by immunohistochemistry and Western blot.Results:Compared with the control group,the body weight of the aging model group and Met group decreased,the weight and index of testis decreased in the aging model group(P<0.01),and increased significantly after Met intervention(P<0.001).The levels of T,FSH and Ein aging model group decreased significantly(P<0.05),and the levels of LH increased significantly(P<0.01).The expression ofβ-gal,p16 and p21 in testis cytoplasm was significantly higher in the model group than in the model group.The expression levels of testosterone synthesis key enzymes,StAR,CYP11 A1 and HSD17β3,significantly increased after Met treatment(P<0.05).The expression of autophagy-related proteins,PINK1,Parkin,Beclin-1 and LC3,significantly increased(P<0.05),and the expression of p62 significantly decreased(P<0.05).Conclusion:Metformin can activate testicular autophagy and increase the level of key enzymes of testosterone synthesis,thus preventing testicular hypofunction in subacute aging rats.

关 键 词：二甲双胍 自噬 亚急性衰老大鼠 睾丸功能 

分 类 号：R965[医药卫生—药理学] R977.6[医药卫生—药学]