强心颗粒通过调控Rac1/NOX/ROS轴抑制慢性心力衰竭心气虚兼血瘀水肿证大鼠心肌细胞凋亡研究  

Study on Qiangxin granules in inhibiting myocardial cell apoptosis in rats with chronic heart failure with heart-qi deficiency leading to blood stasis edema syndrome by regulating the Rac1/NOX/ROS axis

作　　者：李洋 部帅[2] 李慧[3] 王海永 李振祥[5] 林家茂[6] LI Yang;BU Shuai;LI Hui;WANG Haiyong;LI Zhenxiang;LIN Jiamao(Shandong Cancer Hospital and Institute,Shandong First Medical University and Shandong Academy of Medical Sciences,Jinan250017,China;Department of Cardiology,The Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan250001,China;Emergency Department,The Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250011,China)

机构地区：[1]山东省肿瘤防治研究院山东省肿瘤医院放射免疫与分子影像实验室,山东济南250117 [2]山东中医药大学第二附属医院心血管科,山东济南250011 [3]山东中医药大学附属医院急诊科,山东济南250011 [4]山东省肿瘤防治研究院内三科,山东济南250117 [5]山东省肿瘤防治研究院胸部放疗一病区,山东济南250117 [6]山东省肿瘤防治研究院中西医结合病区,山东济南250117

出　　处：《社区医学杂志》2023年第13期658-664,共7页Journal Of Community Medicine

基　　金：国家自然科学基金青年基金(81904186)。

摘　　要：目的检测强心颗粒通过调控Rac1蛋白磷酸化抑制慢性心力衰竭心气虚兼血瘀水肿证活性氧(ROS)诱导心肌细胞凋亡的机制。方法应用多柔比星联合丙硫氧嘧啶(PTU)法复制病证结合心力衰竭大鼠模型;分为正常组、模型组、强心颗粒组以及强心颗粒+Rac1过表达组。干预4周后检测各组大鼠心肌总Rac1、磷酸化Rac1(P-Rac1)以及下游蛋白NOX2、NOX4的表达情况,对比各组心肌ROS含量、8-OHdG核染(+)比例、心肌细胞凋亡指数。结果正常组未死亡,模型组存活率为58.33%,强心颗粒组存活率为72.73%,强心颗粒+Rac1过表达组存活率为72.73%。ROS检测结果显示,模型组心肌细胞ROS荧光强度为34.44±1.63,强心颗粒组为18.39±1.50,强心颗粒+Rac1过表达组为26.23±2.01,强心颗粒组低于模型组(t=20.32,P<0.001),强心颗粒+Rac1过表达组高于强心颗粒组,t=10.04,P<0.01。免疫组化结果显示,模型组心肌细胞8-OHdG核染(+)比率为19.21±2.33,强心颗粒组为11.78±1.26,强心颗粒+Rac1过表达组为17.79±2.00,强心颗粒组低于模型组(t=8.07,P<0.001),强心颗粒+Rac1过表达组高于强心颗粒组,t=8.96,P<0.001。TUNEL检测结果显示,模型组心肌细胞凋亡指数为35.59±3.45,强心颗粒组为14.69±2.66,强心颗粒+Rac1过表达组为29.08±3.20,强心颗粒组低于模型组(t=15.37,P<0.001),强心颗粒+Rac1过表达组高于强心颗粒组t=11.04,P<0.001。蛋白质印迹法检测结果显示,强心颗粒组心肌Rac1、P-Rac1、NOX2和NOX4表达水平均升高,而强心颗粒+Rac1过表达组心肌Rac1、P-Rac1、NOX2和NOX4表达水平均高于强心颗粒组,差异有统计学意义,均P<0.05。结论强心颗粒可显著减少心肌ROS含量,提高心肌Rac1、P-Rac1以及NOX2、NOX4表达水平,减少8-OHdG核染(+)心肌细胞比率,减少心肌细胞凋亡指数,心肌Rac1蛋白磷酸化是强心颗粒抑制ROS诱导的心肌细胞凋亡的重要调控机制,对慢性心力衰竭心气虚兼血瘀水肿证Rac1/NOX/ROS轴具�Objective To investigate the mechanism of Qiangxin granules inducing myocardial cell apoptosis by regulating the phosphorylation of Rac1 protein to inhibit reactive oxygen species(ROS)in chronic heart failure with heart-qi deficiency leading to blood stasis edema syndrome.Methods Experimental rat model was established by doxorubicin plus propylthiouracil method.The normal group,model group,Qiangxin granule group and Qiangxin granule plus Rac1 over-expression group were included.After 4 weeks treatment,total Rac1,phosphorylated Rac1(P-Rac1),NOX2 and NOX4,myocardial ROS content,8-OHdG nuclear staining(+)ratio,and cardiomyocytes apoptotic index were evaluated.Results The normal group did not die,the survival rate of the model group was 58.33%,the survival rate of the Qiangxin granule group was 72.73%,and the survival rate of the Qiangxin granule plus Rac1 over-expression group was 72.73%.The ROS detection results showed that the ROS fluorescence intensity of myocardial cells in the model group was 34.44±1.63,the Qiangxin granule group was 18.39±1.50,and the Qiangxin granule plus Rac1 over-expression group was 26.23±2.01.The Qiangxin granule group was lower than the model group(t=20.32,P<0.001),and the Qiangxin granule plus Rac1 over-expression group was higher than the Qiangxin granule group(t=10.04,P<0.01).The immunohistochemical results showed that the 8-OHdG nuclear staining(+)ratio of myocardial cells in the model group was 19.21±2.33,the Qiangxin granule group was 11.78±1.26,and the Qiangxin granule plus Rac1 over-expression group was 17.79±2.00.The Qiangxin granule group was lower than the model group(t=8.07,P<0.001),and the Qiangxin granule plus Rac1 over-expression group was higher than the Qiangxin granule group(t=8.96,P<0.001).The TUNEL test results showed that the myocardial cell apoptosis index in the model group was 35.59±3.45,the Qiangxin granule group was 14.69±2.66,and the Qiangxin granule plus Rac1 over-expression group was 29.08±3.20.The Qiangxin granule group was lower than the model grou

关 键 词：强心颗粒 慢性心力衰竭 心气虚兼血瘀水肿证 心肌凋亡 Rac1蛋白磷酸化 

分 类 号：R285.5[医药卫生—中药学]