滋膵通脉饮对高糖诱导大鼠H9c2心肌细胞自噬和凋亡的影响及机制  

Effect and mechanism of Zicui Tongmai Yin on autophagy and apoptosis of H9c2 cardiomyocytes induced by high glucose in rats

作　　者：吴刚强[1] 袁春云[1] 毛叶[1] 陈琪[1] 温小凤[1] 谭军[1] 卜献春[1] 李非[2] WU Gangqiang;YUAN Chunyun;MAO Ye;CHEN Qi;WEN Xiaofeng;TAN Jun;BU Xianchun;LI Fei(Departmend of Elderly Cadres,Inheritance Studio of Bu Xianchun,a Famous and Old Chinese Medicine Expert in Affiliated Hospital of Hunan Academy of Chinese Medicine,Changsha 410006,Hunan Province,China;Department of Pharmacy,Affiliated Hospital of Hunan Academy of Chinese Medicine,Changsha 410006,Hunan Province,China)

机构地区：[1]湖南省中医药研究院附属医院老干科/全国名老中医药专家卜献春传承工作室,湖南长沙410006 [2]湖南省中医药研究院附属医院药剂科,湖南长沙410006

出　　处：《新乡医学院学报》2023年第6期507-514,共8页Journal of Xinxiang Medical University

基　　金：湖南省中医药管理局一般项目(编号:2021071);长沙市自然科学基金项目(编号:Kq2202476)。

摘　　要：目的 探讨滋膵通脉饮对高糖诱导的大鼠H9c2心肌细胞自噬和凋亡的影响及机制。方法 将20只无特定病原级Sprague Dawley大鼠按随机数字表法分为空白血浆组和滋膵通脉饮组,每组10只。滋膵通脉饮组大鼠每天给予10 mL·kg^(-1)滋膵通脉饮灌胃,空白血浆组大鼠给予等剂量灭菌超纯水灌胃,连续灌胃7 d,采集腹主动脉血,分离血浆备用。取对数生长期H9c2心肌细胞,按随机数字表法分为正常对照组、空白对照组和滋膵通脉饮组,分别加入稀释后的胎牛血清、空白血浆组血浆及滋膵通脉饮组血浆,每组设置体积分数5%、10%、15%的浓度梯度,应用细胞计数试剂盒-8法检测各组H9c2心肌细胞增殖情况,筛选最佳含药血浆浓度。取对数生长H9c2细胞,按随机数字表法分为正常对照组、高糖诱导组、高糖诱导+中药含药血浆组、高糖诱导+恩格列净组,正常对照组细胞不加任何药物干预,高糖诱导组细胞加33.3 mmol·L^(-1) D-葡萄糖和体积分数10%空白血浆组血浆,高糖诱导+中药含药血浆组细胞加33.3 mmol·L^(-1) D-葡萄糖和体积分数10%滋膵通脉饮组血浆,高糖诱导+恩格列净组细胞加33.3 mmol·L^(-1)的D-葡萄糖和0.01μmol·L^(-1)恩格列净;给药24 h后,使用细胞毒性比色法检测各组细胞乳酸脱氢酶(LDH)释放率,流式细胞术检测各组细胞凋亡率,Western blot法检测各组细胞中自噬和凋亡相关蛋白p62、Bcl-2相关X蛋白(Bax)、微管相关蛋白1轻链3(LC3)-Ⅰ、LC3-Ⅱ和Bcl-2的表达。结果 体积分数5%、15%浓度时,滋膵通脉饮组H9c2细胞增殖能力显著低于正常对照组和空白对照组(P<0.05);正常对照组与空白对照组H9c2细胞增殖能力比较差异无统计学意义(P>0.05)。体积分数10%浓度时,正常对照组、空白对照组和滋膵通脉饮组H9c2细胞增殖能力比较差异无统计学意义(F=0.110,P>0.05)。正常对照组、高糖诱导+中药含药血浆组和高糖诱导+恩格列净Objective To investigate the effect and mechanism of Zicui Tongmai Yin on autophagy and apoptosis of H9c2 cardiomyocytes induced by high glucose in rats.Methods Twenty free specific pathogenic grade Sprague Dawley rats were randomly divided into blank plasma group and Zicui Tongmai Yin group by using a random number table method,with 10 rats in each group.The rats in the Zicui Tongmai Yin group were given 10 mL·kg^(-1) Zicui Tongmai Yin by gavage every day,for 7 days;and the rats in the blank plasma group were given equal dose of sterilized ultrapure water by gavage for 7 days.Abdominal aortic blood was collected and plasma was separated for standby.The H9c2 cardiomyocytes in logarithmic growth phase were divided into normal control group,blank control group and Zicui Tongmai Yin group according to random number table method;and the diluted fetal bovine serum,plasma in the blank plasma group and plasma in the Zicui Tongmai Yin group were added in each group,and concentration gradients of 5%,10%and 15%were set in each group.The proliferation of H9c2 cardiomyocytes in each group was detected by using cell counting kit-8 method to screen the optimal drug containing plasma concentration.The H9c2 cardiomyocytes in logarithmic growth phase were randomly divided into normal control group,high-glucose induction group,high-glucose induction+traditional Chinese medicine containing plasma group and high-glucose induction+empagliflozin group by using a random number table method.The cells in the normal control group did not receive any drug intervention,while the cells in the high-glucose induction group were added 33.3 mmol·L^(-1) of D-glucose and 10%plasma in the blank control group,the cells in the high-glucose induction+traditional Chinese medicine plasma group were added 33.3 mmol·L^(-1) of D-glucose and 10%plasma in the Zicui Tongmai Yin group,and the cells in the high-glucose induction+empagliflozin group cells were treated with 33.3 mmol·L^(-1) D-glucose and 0.01μmol·L-1 empagliflozin;after 24 hours of administ

关 键 词：滋膵通脉饮 H9C2心肌细胞 自噬 凋亡 

分 类 号：R285.5[医药卫生—中药学]