温阳振衰颗粒调控miR-132-3p/UCP2表达对脓毒症大鼠心肌细胞自噬、凋亡的影响  

Effect of Wenyang Zhenshuai Granules on autophagy and apoptosis of myocardial cells in septic rats via regulating miR-132-3p/UCP2 expression

作　　者：王建湘[1] 卢青[1] 陈青扬[1] 廖杨 易琼[1] WANG Jian-xiang;LU Qing;CHEN Qing-yang;LIAO Yang;YI Qiong(Department of Critical Care Medicine,the First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China)

机构地区：[1]湖南中医药大学第一附属医院重症医学科,湖南长沙410007

出　　处：《中国中药杂志》2023年第11期3066-3073,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82204986);湖南省自然科学基金项目(2021JJ40426);湖南省中医药科研计划项目(E2022029)。

摘　　要：探讨温阳振衰颗粒调控微小核糖核酸-132-3p(microribonucleic acid-132-3p,miR-132-3p)/解偶联蛋白2(uncoupling protein 2,UCP2)表达对脓毒症大鼠心肌细胞自噬、凋亡的影响。取60只SD大鼠随机分为模型组(50只)和假手术组(10只)。模型组大鼠通过盲肠结扎穿孔术构建脓毒症大鼠模型,将建模成功的大鼠随机分为温阳振衰颗粒低、中、高剂量组,模型组,阳性对照组。假手术组大鼠仅开腹游离盲肠不进行穿孔结扎。苏木素-伊红(hematoxylin-eosin,HE)染色观察大鼠心肌组织病理学改变;原位末端标记法(TdT-mediated dUTP nick end labeling,TUNEL)检测大鼠心肌细胞凋亡;RT-qPCR检测大鼠心肌组织miR-132-3p表达、UCP2、微管相关蛋白轻链3(microtubule-associated protein light chain 3,LC3-Ⅱ/LC3-Ⅰ)、Beclin-1、半胱氨酸蛋白酶-3(caspase-3)mRNA表达;Western blot检测大鼠心肌组织UCP2、LC3-Ⅱ/LC3-Ⅰ、Beclin-1、caspase-3蛋白表达;双荧光素酶报告基因实验验证miR-132-3p与UCP2的调控关系。脓毒症模型大鼠心肌纤维排列紊乱,有明显炎症细胞浸润和心肌细胞水肿、坏死现象。随温阳振衰颗粒剂量的增加,心肌组织病理学改变均得到不同程度的改善。与假手术组比,模型组,阳性对照组,温阳振衰颗粒低、中、高剂量组大鼠生存率及LVEF降低,心肌损伤评分和细胞凋亡率升高;与模型组比,阳性对照组及温阳振衰颗粒低、中、高剂量组大鼠生存率及LVEF升高,心肌损伤评分和细胞凋亡率降低。与假手术组比,模型组,阳性对照组,温阳振衰颗粒低、中、高剂量组大鼠心肌组织miR-132-3p、UCP2 mRNA表达及蛋白表达降低,LC3-Ⅱ/LC3-Ⅰ、Beclin-1、caspase-3 mRNA及蛋白表达升高;与模型组比,阳性对照组及温阳振衰颗粒低、中、高剂量组大鼠心肌组织miR-132-3p表达、UCP2 mRNA及蛋白表达升高,LC3-Ⅱ/LC3-Ⅰ、Beclin-1、caspase-3 mRNA及蛋白表达降低。温阳振衰颗粒可抑制脓毒症�This study aimed to investigate the effect of Wenyang Zhenshuai Granules(WYZSG)on autophagy and apoptosis of myocardial cells in rats with sepsis via regulating the expression of microRNA-132-3p(miR-132-3p)/uncoupling protein 2(UCP2).Sixty SD rats were randomly divided into modeling group(n=50)and sham operation group(n=10).The sepsis rat model was constructed by cecal ligation and perforation in the modeling group.The successfully modeled rats were randomly divided into WYZSG low-,medium-and high-dose groups,model group and positive control group.Rats in the sham operation group underwent opening and cecum division but without perforation and ligation.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of rat myocardial tissue.Myocardial cell apoptosis was detected by TdT-mediated dUTP nick end labeling(TUNEL)assay.Real-time quantitative polymerase chain reaction(RT-qPCR)was performed to detect the expression of miR-132-3p and the mRNA expressions of UCP2,microtubule-associated protein light chain 3(LC3-Ⅱ/LC3-Ⅰ),Beclin-1 and caspase-3 in rat myocardial tissue.The protein expressions of UCP2,LC3-Ⅱ/LC3-Ⅰ,Beclin-1 and caspase-3 in myocardial tissue were detected by Western blot.Dual luciferase reporter assay was used to verify the regulatory relationship between miR-132-3p and UCP2.The myocardial fibers of sepsis model rats were disordered,and there were obvious inflammatory cell infiltration as well as myocardial cell edema and necrosis.With the increase of the WYZSG dose,the histopathological changes of myocardium were improved to varying degrees.Compared with the conditions in the sham operation group,the survival rate and left ventricular ejection fraction(LVEF)of rats in the model group,positive control group and WYZSG low-,medium-and high-dose groups were decreased,and the myocardial injury score and apoptosis rate were increased.Compared with the model group,the positive control group and WYZSG low-,medium-and high-dose groups had elevated survival rate and LVEF,and lowered myo

关 键 词：温阳振衰颗粒 miR-132-3p 解偶联蛋白2(UCP2) 脓毒症 心肌细胞 自噬 凋亡 

分 类 号：R285.5[医药卫生—中药学]