积雪草苷调控GRP78/PERK/CHOP通路对急性心肌梗死大鼠的心肌保护作用研究  

Study on Protective Effects of Asiaticoside on GRP78/PERK/CHOP Pathway in Rats with Acute Myocardial Infarction

作　　者：杨勇[1] 殷红珍[2] 胡云飞 Yang Yong;Yin Hongzhen;Hu Yunfei(Chuzhou First People's Hospital,Anhui,Chuzhou 239000,China)

机构地区：[1]安徽省滁州市第一人民医院,安徽滁州239000 [2]皖南医学院弋矶山医院,安徽芜湖241000 [3]皖南医学院,安徽芜湖241000

出　　处：《中国中医急症》2019年第9期1571-1575,共5页Journal of Emergency in Traditional Chinese Medicine

基　　金：安徽省自然科学基金项目(1708084MH027)

摘　　要：目的观察积雪草苷(AC)通过内质网应激(ERS)相关葡萄糖调节蛋白78(GRP78)/蛋白激酶R样内质网激酶(PERK)/转录因子C、EBP同源蛋白(CHOP)通路对急性心肌梗死(AMI)大鼠的心肌保护作用。方法选择SPF级8周龄级SD大鼠108只,结扎左冠状动脉前降支建立心肌梗死模型,随机分为假手术组,模型组,积雪草苷低、中、高剂量组,牛磺酸组,造模完成24h后给药,每天早晚各1次,4周后测定左室质量和体质量,TCC染色法测定心肌梗死面积,TUNEL法检测心肌细胞凋亡率,HE染色观察心肌细胞形态,Westorn blotting检测GRP78、PERK、CHOP及凋亡相关蛋白Bax、Bcl-2、Caspase-12、Caspase-3的表达。结果与假手术组比较,模型组左室质量和左室质量/体质量的比值显著增加(P<0.05),心肌梗死面积显著增加,心肌细胞凋亡率显著增加;与模型组比较,积雪草苷各组和牛磺酸组左室质量和左室质量/体质量的比值显著下降(P<0.05),梗死面积下降,细胞凋亡比例显著减少(P<0.05),且积雪草苷各组呈浓度依赖性降低。HE染色显示假手术组心肌纤维排列整齐,横纹清晰,无水肿或坏死;模型组心肌纤维不规则,横纹不清或缺失,积雪草苷各组和牛磺酸组心肌纤维轻度扩张,心肌纤维局灶性丢失。Western blotting检测显示,与假手术组比较,模型组GRP78、PERK、CHOP、Bax、Caspase-12、Caspase-3蛋白表达水平显著升高(P<0.05),Bcl-2蛋白水平显著降低(P<0.05);与模型组比较,积雪草苷各组随着浓度的升高GRP78、p-PERK、CHOP、Bax、Caspase-12、Caspase-3蛋白表达逐渐降低(P<0.05),Bcl-2蛋白表达逐渐升高(P<0.05),均呈剂量依赖性。结论AC可减少大鼠心肌梗死区,可通过抑制GRP78/PERK/CHOP通路发挥对急性心肌梗死大鼠的保护作用。Objective:To investigate the protective effects of asiaticoside(AC)on myocardium in rats with AMI through ERS related GRP78/PERK/transcription factor C,EBP homologous protein(CHOP) pathway.Methods: 108 SPF 8-week-old SD rats were selected to establish myocardial infarction model by ligating left anterior descending coronary artery,and randomly divided into the sham group,the model group(AMI),asiaticoside low,medium and high dose groups(Asiaticoside-L,Asiaticoside-M,Asiaticoside-H)and taurine group(Tau group).Drug was given from the day of modeling,once in the morning and in the evening.Left ventricular weight and body weight were measured after 4 weeks;the area of myocardial infarction was measured by TCC staining;the apoptotic rate of myocardial cells was detected with TUNEL method;HE staining was used to observe the morphology of myocardial cells;the expressions of GRP78,PERK,CHOP and apoptosis-related proteins Bax,Bcl-2,Caspase-12 and Caspase-3 were detected by Westorn blotting.Results:Compared with the sham group,the left ventricular weight and the ratio of left ventricular weight to body weight in AMI group increased significantly(P < 0.05);the area of myocardial infarction(45.68%)increased significantly;the apoptotic rate of cardiomyocytes increased significantly.Compared with AMI group,the left ventricular weight and the ratio of left ventricular weight to body weight decreased significantly in Asiaticoside groups and Tau group(P < 0.05);the infarct size decreased;the percentage of apoptotic cells decreased significantly(P < 0.05);the siaticoside decreased in a concentration-dependent manner.HE staining showed that the myocardial fibers in the sham group were well arranged;the lines were clear,no edema or necrosis;the muscle fibers in the AMI group were irregular;striations were lack or absence;the muscle fibers of Asiaticoside group and Tau group were slightly dilated;muscle fibers were focal lost.Western blotting showed that the expression levels of GRP78,p-PERK,CHOP,Bax,Caspase-12 and Caspase-3 proteins

关 键 词：急性心肌梗死 积雪草苷 内质网应激 GRP78 PERK 大鼠 

分 类 号：R285.5[医药卫生—中药学]