参芪复方对GK大鼠骨骼肌线粒体膜电位及相关促凋亡蛋白的影响研究  

Effects of Shenqi Compound Prescription on Skeletal Muscle Mitochondrial Membrane Potential and Related Pro-apoptotic Proteins in GK Rats

作　　者：曹纪辉 吴贤波 丁若兰[3] 冯树贵 朱海燕 Cao Jihui;Wu Xianbo;Ding Ruolan;Feng Shugui;Zhu Haiyan(College of Clinical Medicine,Chengdu University of Traditional Chinese Medicine,Chengdu 610037,China;College of Sports Medicine and Health,Chengdu Sport University,Chengdu 571199,China;Classic Ward of TCM,Affiliated Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610072,China;Clinical Medical College and The First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都中医药大学临床医学院,成都610037 [2]成都体育学院运动医学与健康学院,成都610041 [3]成都中医药大学附属医院,成都610072 [4]成都医学院临床医学院第一附属医院,成都610500

出　　处：《世界科学技术-中医药现代化》2021年第7期2302-2308,共7页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委员会地区科学基金项目(81660788):从线粒体动力学研究益气健脾活血法干预T2DM骨骼肌细胞凋亡的作用机制,负责人:朱海燕;四川省中医药管理局科学技术研究专项(2018QN051):基于肠道菌群研究益气健脾活血法改善T2DM骨骼肌IR的机制,负责人:朱海燕;成都医学院第一附属医院高层次人才科研启动基金资助项目(CYFY-GQ02):从线粒体动力学研究益气健脾活血法干预T2DM骨骼肌细胞凋亡的作用机制,负责人:朱海燕。

摘　　要：目的探讨参芪复方对自发性糖尿病(Goto-Kakizaki,GK)大鼠骨骼肌线粒体膜电位及相关凋亡蛋白的影响。方法采用高脂饲料喂养GK大鼠,建立糖尿病大鼠动物模型,造模成功后将GK大鼠随机分为模型组、罗格列酮组、参芪复方组,每组10只;10只Wistar大鼠作为空白对照组,普通饲料喂养。罗格列酮组灌胃给予0.67 mg/(kg·d)罗格列酮混悬液,参芪复方组灌胃给予14.33 g生药/(kg·d)参芪复方混悬液,空白对照组、模型组均每日灌胃给予同等体积生理盐水,连续给药8周。采用流式细胞检测线粒体膜电位;RT-qPCR和Western blot检测大鼠腓肠肌凋亡诱导因子(Aapoptosis inducing factor,AIF)、细胞色素C(Cytochrome c,Cyt c)、第二个线粒体衍生的半胱天冬蛋白酶激活因子(Second mitochondrial activator of caspase/direct IAP binding protein with low PI,Smac/DIABLO)、半胱氨酸天冬氨酸蛋白酶-9(Cysteinyl aspartate specific proteinase-9,Caspase-9)、半胱氨酸天冬氨酸蛋白酶-3(Cysteinyl aspartate specific proteinase-3,Caspase-3)的蛋白和mRNA表达情况。结果与空白对照组比较,模型组GK大鼠腓肠肌线粒体膜电位降低(P <0.05),AIF、Cyt c、Smac/DIABLO、Caspase-9、Caspase-3蛋白表达、mRNA表达增加(P <0.05);与模型组比较,罗格列酮组和参芪复方组线粒体膜电位均升高(P <0.05),罗格列酮组AIF、Cyt c、Smac/DIABLO、Caspase-9、Caspase-3蛋白表达、mRNA表达均降低(P <0.05),参芪复方组AIF、Cyt c、Caspase-9、Caspase-3蛋白表达、mRNA表达降低(P <0.05),Smac/DIABLO蛋白表达降低(P <0.05),Smac/DIABLO mRNA表达具有下降趋势,但差异无统计学意义(P> 0.05)。结论参芪复方可通过上调线粒体膜电位,抑制促凋亡蛋白AIF、Cyt c、Smac/DIABLO从线粒体内释放出来,进而抑制Caspase-9、Caspase-3表达,减少骨骼肌细胞凋亡,发挥骨骼肌保护作用。Objective To explore the effect of Shenqi compound prescription on skeletal muscle mitochondrial membrane potential and related apoptosis proteins in Goto-Kakizaki(GK) rats.Methods GK rats were fed with high-fat diet to establish an animal model of diabetic rats. After successful modeling, GK rats were randomly divided into model group, rosiglitazone group, and Shenqi compound group, with 10 rats in each group;10 Wistar rats were served as a blank control group and were fed with ordinary feed. The rosiglitazone group was given 0.67 mg/(kg·d) rosiglitazone suspension by gavage, and the Shenqi compound group was given 14.33 g crude drug/(kg·d) Shenqi compound suspension by gavage, the blank control group and the model group was given an equal volume of normal saline by gavage every day for 8 weeks. Flow cytometry was used to detect mitochondrial membrane potential;RT-qPCR and Western blot were used to detect rat gastrocnemius apoptosis-inducing factor(AIF), cytochrome C(Cyt c), and the second mitochondrial activator of caspase/direct IAP binding protein with low PI(Smac/DIABLO), cysteinyl aspartate specific proteinase-9(Caspase-9), cysteinyl aspartate specific proteinase-3(Caspase-3) protein and mRNA expression.Results Compared with the blank control group, the mitochondrial membrane potential of gastrocnemius muscle of GK rats in the model group decreased(P < 0.05), and the protein expression and mRNA expression of AIF, Cyt c, Smac/DIABLO, Caspase-9 and Caspase-3 increased(P < 0.05);Compared with the model group, the mitochondrial membrane potential of the rosiglitazone group and the Shenqi compound group increased(P < 0.05), and the protein expression of AIF, Cyt c, Smac/DIABLO, Caspase-9, Caspase-3 and mRNA expression in the rosiglitazone group decreased(P <0.05). The expression of AIF, Cyt c, Caspase-9, and Caspase-3 protein of Shenqi compound group decreased(P < 0.05),and the expression of Smac/DIABLO protein decreased(P < 0.05). The Smac/DIABLO mRNA expression had a downward trend, but the difference was no

关 键 词：参芪复方 糖尿病 骨骼肌病变 凋亡 

分 类 号：R284[医药卫生—中药学]