益气活血方治疗心力衰竭的内质网应激机制研究

Study on endoplasmic reticulum stress mechanism of treating heart failure with Yiqi Huoxue Recipe

目的探讨益气活血方通过调节内质网应激(ERS)信号通路改善心力衰竭的分子机制。方法SPF级雄性SD大鼠30只,根据随机数字表法分为假手术组(10只),只开胸不结扎;余20只采用结扎左冠状动脉前降支的方法制备大鼠急性心肌梗死模型。将造模成功的大鼠随机分为模型组、益气活血组,每组10只。模型组和假手术组灌胃去离子水5 mL/(kg·d),1次/d。益气活血组灌胃益气活血方5 mL/(kg·d),连续给药4周。左心室插管检测血流动力学指标包括左心室舒张终末压(LVEDP)、左心室内压最大上升速率(+dp/dt max)、左心室内压最大下降速率(-dp/dt max)、左心室收缩压(LVSP)和心率(HR)。Rt-PCR法检测脑钠肽(BNP)、钙调神经磷酸酶A(CaNA)基因的表达水平,Western blot法检测caspase-12、蛋白激酶R样内质网激酶(PERK)、肌醇需要酶1(IRE1)、葡萄糖调节蛋白78(GRP78)蛋白的表达水平。结果模型组大鼠LVEDP、-dp/dt max、HR高于假手术组;+dp/dt max、LVSP低于假手术组,差异有高度统计学意义(P<0.01)。益气活血组大鼠LVEDP、-dp/dt max、HR低于模型组;LVSP、+dp/dt max高于模型组,差异有高度统计学意义(P<0.01)。模型组大鼠BNP、CaNA mRNA表达高于假手术组,差异有统计学意义(P<0.05或P<0.01)。益气活血组大鼠BNP、CaNA mRNA表达低于模型组,差异有统计学意义(P<0.05)。模型组大鼠caspase-12、PERK、IRE1和GRP78蛋白表达水平高于假手术组,差异有统计学意义(P<0.05);益气活血组大鼠caspase-12、PERK和IRE1蛋白表达水平低于模型组,差异有统计学意义(P<0.05)。结论益气活血方可以改善心肌梗死后大鼠的心功能,其机制与调节ERS信号通路有关。

Objective To explore the molecular mechanism of improvement effect of Yiqi Huoxue Recipe in heart failure treatment by regulating endoplasmic reticulum stress signaling(ERS)pathway.Methods Thirty SPF male SD rats were divided into sham operation group(10 rats)by the random number table method,and the remaining 20 rats model of acute myocardial infarction was prepared by ligating the anterior descending branch of the left coronary artery.Rats with successful modeling were randomly divided into model group and Yiqi Huoxue group,with 10 rats in each group.The rats in model group and sham operation group were gavaged with deionized water 5 mL/(kg·d),1 time/d.Rats in Yiqi Huoxue group were gavaged with Yiqi Huoxue Recipe 5 mL/(kg·d).The drug was administered continuously for 4 weeks.Hemodynamic index including left ventricular end-diastolic pressure(LVEDP),left ventricular internal pressure maximal rate of increase(+dp/dt max),left heart maximum drop rate of indoor pressure(-dp/dt max),left ventricular systolic pressure(LVSP)and heart rate(HR)were detected by left ventricular intubation method.The expressions of brain natriuretic peptide(BNP)and calcineurin A(CaNA)mRNA were detected by Rt-PCR and the protein levels of caspase-12,protein kinase R-like endoplasmic reticulum kinase(PERK),inositol requiring enzymes 1(IRE1)and glucose-regulated protein 78(GRP78)were detected by Western blot.Results LVEDP,-dp/dt max and HR in model group were higher than those in sham group,+dp/dt max and LVSP were significantly lower than those in sham operation group,and the differences were highly statistically significant(P<0.01).LVEDP,-dp/dt max and HR of rats in Yiqi Huoxue group were lower than those in model group,LVSP and+dp/dt max were higher than those in model group,and the differences were highly statistically significant(P<0.01).The expressions of BNP and CaNA mRNA in model group were higher than those in sham operation group,with statistically significant differences(P<0.05 or P<0.01).The expressions of BNP and CaNA mRNA in Yiqi Huoxue group were lower than those in model group,with statistically significant differences(P<0.05).The expression levels of caspase-12,PERK,IRE1 and GRP78 in model group were higher than those in sham operation group,with statistically significant differences(P<0.05).The expression levels of caspase-12,PERK and IRE1 in Yiqi Huoxue group were lower than those in model group,with statistically significant differences(P<0.05).Conclusion Yiqi Huoxue Recipe can improve the cardiac function of rats with heart failure induced by myocardial infarction,and the mechanism included may be related to the endoplasmic reticulum stress signaling pathway regulation.