两和辅心饮通过NRG1-ErbB2通路对心肌细胞损伤的干预研究

Effect of Lianghe Fuxin Potions on Myocardial Cell Injury through NRG1-ErbB2 Pathway

目的研究两和辅心饮通过神经调节蛋白1(NRG1)/erb-b2受体酪氨酸蛋白激酶(ErbB)2通路对液氮冷冻大鼠心肌梗死模型中心肌细胞损伤的影响,探讨该方治疗心肌梗死的作用机制。方法无特定病原体(SPF)级Wistar雄性大鼠80只随机分为5组,假手术组(Sham组)、模型组(MIRI组)、低剂量组、中剂量组、高剂量组。心肌梗死模型造模成功后予高、中、低剂量组相应剂量两和辅心饮灌胃(生药量分别为0.96 g/100 g、1.92 g/100 g、3.84 g/100 g),Sham组、模型组给予等量蒸馏水,给药周期为3周。实验结束后取大鼠心肌组织,苏木素-伊红(HE)染色观察心肌组织病理变化;蛋白质免疫印迹法(Western Blot)测定心肌组织中NRG1、ErbB2的蛋白表达水平;实时荧光定量聚合酶链式反应(RT-PCR)测定大鼠心肌细胞中NRG1、ErbB2的mRNA表达水平。结果①HE染色显示,经过两和辅心饮治疗后,心肌细胞排列逐渐整齐,心肌细胞的肿胀程度明显减轻,间质充血减轻,炎性细胞浸润有所缓解。②Western Blot测定结果显示:与Sham组比较,MIRI组NRG1、ErbB2蛋白表达均降低,差异有统计学意义(P<0.05);低、中、高剂量组NRG1及ErbB2蛋白表达较模型组升高,并呈药物剂量依赖性(P<0.05)。灰度分析显示:与MIRI组比较,低剂量组、中剂量组、高剂量组NRG1/3-磷酸甘油醛脱氢酶(GAPDH)、ErbB2/GAPDH表达水平升高,并呈剂量依赖性(P<0.05)。③RT-PCR检测结果显示:与Sham组相比,MIRI组NRG1、ErbB2的mRNA表达明显降低,显示出随着心肌缺血缺氧的发生,NRG1、ErbB2在心肌中的含量降低;低剂量组、中剂量组、高剂量组NRG1、ErbB2的mRNA表达较MIRI组明显升高,且剂量越高mRNA表达越接近于Sham组,呈剂量依赖性。结论两和辅心饮可能通过激活NRG1-ErbB2通路修复大鼠心肌细胞损伤。

Objective To study the effect of Lianghe Fuxin Potions on neuregulin-1(NRG1)/erb-b2 receptor tyrosine kinase(ErbB)2 pathway of myocardial cell in rats with myocardial infarction induced by liquid nitrogen freezing,and explore its mechanism of action in the treatment of myocardial infarction.Methods Eighty SPF grade male Wistar rats were randomly divided into 5 groups:sham operation group,model group(MIRI group),low-dose group,medium dose group,and high-dose group.After the successful establishment of myocardial infarction model,the rats in high-dose group,medium-dose group,and low-dose group were given Lianghe Fuxin Potions by gavage(crude drug dosage:0.96 g/100 g,1.92 g/100 g,and 3.84 g/100 g,respectively).The rats in sham operation group and model group were given equal amount of distilled water,and the experimental period was 3 weeks.At the end of the experiment,the pathological changes of myocardial tissue were observed by hematoxylin-eosin(HE)staining.The expression levels of NRG1 and ErbB2 in myocardial tissue were detected by Western Blot.The mRNA expression levels of NRG1 and ErbB2 in myocardial cells were detected by fluorescence quantitative polymerase chain reaction(PCR).Results①HE staining showed that after treatment with Lianghe Fuxin Potions,cardiomyocytes were gradually arranged in order,the swelling of cardiomyocytes was significantly reduced,interstitial congestion was reduced,and inflammatory cell infiltration was alleviated.②Western Blot results showed that compared with sham operation group,the expression of NRG1 and ErbB2 protein in MIRI group was decreased significantly(P<0.05).The protein expressions of NRG1 and ErbB2 in low,medium,and high dose groups were higher than those in model group,and they were positively correlated with drug dose(P<0.05).Gray analysis showed that the expression levels of NRG1/GAPDH and ErbB2/GAPDH in low-dose group,medium-dose group,and high-dose group were increased compared with that in MIRI group,and they were positively correlated with drug dose(P<0.05).③Fluorescence quantitative PCR results showed that compared with sham operation group,the mRNA expression of NRG1 and ErbB2 in MIRI group was decreased significantly,showing that the contents of NRG1 and ErbB2 in myocardium was decreased with the occurrence of myocardial ischemia and hypoxia.The expressions of NRG1 and ErbB2 mRNA in low-dose group,medium-dose group,and high-dose group were significantly higher than those in MIRI group,and the higher the dose,the closer the mRNA expression in sham group,with a positive correlation with the dose.Conclusion Lianghe Fuxin Potions may repair myocardial cell injury in rats by activating NRG1-ErbB2 pathway.