基于HIFs/RhoB/ROCK通路探讨黄芪甲苷改善缺氧诱导心力衰竭大鼠心肌损伤的作用机制  

The Mechanism of Astragaloside on Myocardium Injury in Rats of Hypoxic-induced Heart Failure Based on HIFs/RhoB/ROCK Pathway

作　　者：张佳[1] 秦蕊 李刚 ZHANG Jia;QIN Rui;LI Gang(Hubei University of Chinese medicine,Wuhan 430065,Hubei,China;Affiliated Hospital of Hubei University of Chinese Medicine,Wuhan 430074,Hubei,China;Hubei Provincal Hospital of Traditional Chinese Medicine,Wuhan 430074,Hubei,China)

机构地区：[1]湖北中医药大学,武汉430065 [2]湖北中医药大学附属医院,武汉430074 [3]湖北省中医院,武汉430074

出　　处：《中西医结合心脑血管病杂志》2025年第6期863-868,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：湖北省自然科学基金创新发展联合基金项目(No.2022CFD158)。

摘　　要：目的:观察黄芪甲苷(AST)对缺氧诱导的心力衰竭大鼠血流动力学相关指标、心肌肌钙蛋白及细胞凋亡的改善作用,并探讨其通过调节缺氧诱导因子(HIFs)/RhoB蛋白(RhoB)/Rho相关激酶(ROCK)信号通路的改善机制。方法:采用大鼠尾静脉注射盐酸阿霉素建立心力衰竭大鼠模型,AST治疗后,再包装HIF-1α-干扰腺病毒(HIF-1αsiRNA)和HIF-2α-干扰腺病毒(HIF-2αsiRNA)干预大鼠。将大鼠分为假手术组(Sham组)、心力衰竭组(HF组)、心力衰竭+AST组(HF+AST组)、心力衰竭+AST+阴性对照组(HF+AST+NC组)、心力衰竭+AST+siRNA HIF-1α组(HF+AST+siRNA HIF-1α组)及心力衰竭+AST+siRNA HIF-2α组(HF+AST+siRNA HIF-2α组)。测定大鼠心肌肌钙蛋白(I cTnI)、心率、收缩压、舒张压及平均血压(MBP)变化;通过流式细胞术检测活性氧变化。通过实时荧光定量聚合酶链式反应(qRT-PCR)检测HIF-1α、HIF-2αmRNA表达水平;蛋白免疫印记法(Western Blot)检测HIF-1α、HIF-2α、RhoB、ROCK表达变化;酶联免疫吸附法(ELISA)检测Caspase-3表达水平。结果:与HF组比较,HF+AST组大鼠心率、舒张压、MBP、cTnI、活性氧、Caspase-3水平下降,心肌细胞HIF-1α、HIF-2αmRNA与蛋白表达升高,RhoB和ROCK蛋白表达升高(P<0.05);与HF+AST+NC组比较,HF+AST+siRNA HIF-1α组及HF+AST+siRNA HIF-2α组大鼠心率、收缩压、MBP升高,cTnI、活性氧、Caspase-3水平升高,心肌细胞RhoB、ROCK蛋白表达下降(P<0.05)。结论:AST可减轻缺氧诱导的心力衰竭模型大鼠心肌细胞损伤,其机制可能与HIFs/RhoB/ROCK通路相关。Objective:To observe the effect of astragaloside(AST)on hemodynamic parameters,troponin and apoptosis in hypoxiainduced heart failure rats,and to explore the effect of AST by regulating hypoxia inducible factor(HIFs)/RhoB protein(RhoB)/Rhoassociated kinase(ROCK)signaling pathway.Methods:The rat model of heart failure was established by injecting adriamycin hydrochloride into the tail vein.After AST treatment,the rats were treated with HIF-1α-interfering adenovirus(HIF-1αsiRNA)and HIF-2α-interfering adenovirus(HIF-2αsiRNA).The rats were divided into Sham group,heart failure group(HF group),HF+AST group,HF+AST+negative control group(HF+AST+NC group),HF+AST+siRNA HIF-1αgroup(HF+AST+siRNA HIF-1αgroup),heart failure+AST+siRNA HIF-2αgroup(HF+AST+siRNA HIF-2αgroup).The changes of cardiac troponin I(cTnI),heart rate,systolic blood pressure,diastolic blood pressure,and mean blood pressure(MBP)were measured and reactive oxygen species were detected by flow cytometry.The mRNA expressions of HIF-1αand HIF-2αwere detected by Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).The expressions of HIF-1α,HIF-2α,RhoB,and ROCK were detected by Western Blot.The expression level of Caspase-3 was detected by enzyme-linked immunosorbent assay(ELISA).Results:Compared with HF group,the heart rate,diastolic blood pressure,and MBP in the HF+AST group increased,the levels of cTnI,ROS and Caspase-3 decreased,the mRNA and protein expressions of HIF-1αand HIF-2αincreased,and the protein expressions of RhoB and ROCK increased(P<0.05).Compared with HF+AST+NC group,the heart rate,systolic blood pressure and MBP of rats in the HF+AST+siRNA HIF-1αgroup and HF+AST+siRNA HIF-2αgroup increased,and the levels of cTnI,reactive oxygen species and Caspase-3 significantly increased,the expression of RhoB and ROCK protein in cardiomyocytes decreased(P<0.05).Conclusion:AST could attenuate cardiomyocyte injury in rats of hypoxic-induced heart failure,and the mechanism might be related to HIFs/RhoB/ROCK pathway.

关 键 词：心力衰竭 缺氧诱导的心肌细胞损伤 黄芪甲苷 缺氧诱导因子 RhoB蛋白 Rho相关激酶 大鼠 实验研究 

分 类 号：R285.5[医药卫生—中药学]