丹参对异丙肾上腺素诱导的心肌损伤内质网应激的影响

Effect of Salvia miltiorrhiza on endoplasmic reticulum stress induced by isoproterenol-induced myocardial injury

目的探讨丹参对异丙肾上腺素(ISO)诱导的心肌损伤内质网应激的干预效果。方法 32只ICR小鼠随机分为4组,即对照组(Con组),模型组(ISO组),丹参低、高剂量治疗组(SL组、SH组)。除Con组外,给与各组ISO,剂量为2mg/(kg·d)腹腔注射4周诱导心肌损伤;第3周开始治疗组同时给与丹参40mg/(kg·d)、80mg/(kg·d)腹腔注射,造模及给药期间Con组和ISO组注射同等剂量生理盐水。实验4周末处死小鼠,称取体质量、心脏质量,计算心脏质量指数(HW/BW);HE和MASSON染色观察病理学改变;蛋白印迹法、Real-time PCR检测GRP94、IRE1、p-JNK、Caspase-12、Cleaved Caspase-3蛋白及mRNA表达情况。结果与对照组比较,ISO组心脏质量和HW/BW增高(P<0.05);心肌细胞肥大,纤维化程度增强;GRP94、IRE1、p-JNK、Caspase-12、Cleaved Caspase-3蛋白及mRNA表达明显增加(P<0.001)。丹参能降低心脏质量、HW/BW及心肌细胞肥大和纤维化程度,明显降低内质网相关因子表达(P<0.01、0.001),且高剂量治疗效果明显。结论内质网应激参与ISO诱导的心肌损伤,丹参能够降低内质网相关因子GRP94、IRE1、p-JNK、Caspase-12、Cleaved Caspase-3的表达水平,改善心肌损伤,且高剂量治疗效果显著。

Objective To explore the effect of Salvia Miltiorrhiza on the isoproterenol(ISO)induced endoplasmic reticulum stress induced myocardial injury.Methods Thirty-two ICR mice were randomly divided into 4 groups,namely control group(Con group),model group(ISO group),low-dose and high-dose treatment groups of Salvia miltiorrhiza(SL group,SH group,respectively).Each group was given ISO,2 mg/(kg·d)intraperitoneal injection for 4 weeks to induce myocardial injury other than the Con group.At the beginning of the third week,the treatment group was also given intraperitoneal injections of Danshen 40 mg/(kg·d)and 80 mg/(kg·d)during the period of administration and administration,and the Con group and the ISO group were injected with the same dose of normal saline.At the end of the fourth week,the mice were sacrificed,the body mass and heart mass were weighed,and the cardiac mass index(HW/BW)was calculated.HE and MASSON staining was used to observe pathological changes.Western blotting and Real-time PCR were used to detect the protein and mRNA expression of GRP94,IRE1,p-JNK,Caspase-12,and Cleaved Caspase-3.Results Compared with the control group,in the ISO group,the heart weight and HW/BW were increased significantly(P<0.05),cardiomyocyte hypertrophy and fibrosis were also increased,and GRP94,IRE1,p-JNK,Caspase-12,Cleared Caspase-3 protein and mRNA were significantly increased(P<0.001).Salvia miltiorrhiza could reduce heart weight,HW/BW,myocardial cell hypertrophy and fibrosis,significantly reduce the expression of endoplasmic reticulum-related factors(P<0.01,0.001),and the high-dose treatment effect.Conclusion Endoplasmic reticulum stress is involved in ISO-induced myocardial injury.Salvia miltiorrhiza can reduce the expression levels of endoplasmic reticulum-related factors GRP94,IRE1,p-JNK,Caspase-12,and Cleaved Caspase-3,play protective role in myocardial injury,and the high-dose treatment has significant protective effect.