三七皂苷R1调控AMPK/Nrf-2/HO-1信号通路缓解冠心病大鼠心肌损伤的研究

Study on Notoginsenoside R1 Regulates AMPK/Nrf-2/HO-1 Signaling Pathway to Relieve Myocardial Damage in Rats with Coronary Heart Disease

关键词:  三七皂苷R1|冠心病|AMPK/Nrf-2/HO-1信号通路

Key words:  notoginsenoside R1|coronary heart disease|AMPK/Nrf-2/HO-1 signaling pathway

摘要:

目的 研究三七皂苷R1对冠心病模型大鼠心肌损伤的作用及机制。方法 将大鼠随机分为对照组，模型组，三七皂苷R1低、中、高剂量组，每组9只；采用饲喂高脂饲料联合垂体后叶注射液腹腔注射的方法建立冠心病大鼠模型；三七皂苷R1低、中、高剂量组分别灌胃给予50，100和200 mg·kg^-1的三七皂苷R1，每天给药1次，连续4周。HE染色观察心肌损伤，Western blotting检测活化的半胱氨酸蛋白酶-3（caspase-3）、半胱氨酸蛋白酶-9（caspase-9）的表达水平，测定平均动脉压、心率和左室收缩压水平，ELISA检测心损标记肌红蛋白（myoglobin，Mb）、肌酸激酶同工酶（creatine kinase isozyme，CK-MB）和肌钙蛋白（cardiac troponin，cTnΙ）表达水平及炎症因子白介素-6（interleukin-6，IL-6）、白介素-1β（interleukin-1β，IL-1β）、诱导型一氧化氮合酶（inducible nitric oxide synthase，iNOS）和肿瘤坏死因子-α（tumor necrosis factor，TNF-α）含量，试剂盒检测心肌细胞丙二醛（malondialdehyde，MDA）、超氧化物歧化酶（superoxide dismutase，SOD）、乳酸脱氢酶（lactate dehydrogenase，LDH）和谷胱甘肽（glutathione，GSH）表达水平，Western blotting检测腺苷酸活化蛋白激酶（adenosine monophosphate-activeted protein kinase，AMPK）的磷酸化及转录因子NF-E2相关因子（Nrf2）、血红素氧合酶-1（heme oxygenase-1，HO-1）表达水平。结果 三七皂苷R1能降低冠心病大鼠心肌损伤程度，抑制心肌凋亡蛋白caspase-3、caspase-9的活化（P<0.05），上调心脏功能指标平均动脉压、心率和左室收缩压水平（P<0.05），抑制心损标记物CK-MB、cTnΙ和Mb高表达（P<0.05），降低氧化应激指标SOD、GSH、LDH和MDA的含量（P<0.05），抑制炎症因子IL-6、IL-1β、iNOS和TNF-α含量表达上调（P<0.05），上调AMPK/Nrf-2/HO-1信号通路蛋白的表达（P<0.05）。结论 三七皂苷R1能改善冠心病大鼠心肌损伤，抑制心肌细胞凋亡、氧化应激、炎症反应，这与调控AMPK/Nrf-2/HO-1信号通路激活有关。

Abstract:

OBJECTIVE To study the effect and mechanism of notoginsenoside R1 on myocardial injury in rats with coronary heart disease. METHODS Rats were randomly divided into control group, model group, notoginsenoside R1 low, medium and high dose group, 9 rats in each group. Rat model of coronary heart disease was established by intraperitoneal injection of high fat diet combined with posterior pituitary injection. The notoginsenoside R1 low, medium and high dose group were administered with notoginsenoside R1 50, 100 and 200 mg·kg^-1 by gavage once a day for 4 weeks. Myocardial injury was observed by HE staining. The expression of cleaved caspase-3 and caspase-9 was detected by Western blotting. Mean ventricular systolic pressure, heart rate and left ventricular systolic pressure levels were measured. Heart damage markers myoglobin(Mb), creatine kinase isozyme(CK-MB) and cardiac troponin(cTnI) expression levels and inflammatory factors interleukin-6(IL-6), interleukin-1β(IL-1β), inducible nitric oxide synthase(iNOS) and tumor necrosis factor-α(TNF-α) contents were detected by ELISA. Myocardial cell malondialdehyde(MDA), superoxide dismutase(SOD), lactate dehydrogenase(LDH) and glutathione (GSH) expression levels were detected by the kit. Phosphorylation of adenosine monophosphate-activeted protein kinase(AMPK) and transcription factor NF-E2 related factor(Nrf2), heme oxygenase-1(HO-1) expression level was detected by Western blotting. RESULTS Notoginsenoside R1 reduced the degree of myocardial injury in rats with coronary heart disease, inhibited the expression of myocardial apoptosis proteins caspase-3 and caspase-9(P<0.05), up-regulated the levels of cardiac function indicators mean ventricular systolic pressure, heart rate and left ventricular systolic pressure(P<0.05), inhibited the high expression of heart injury markers CK-MB, cTnI and Mb(P<0.05), reduced the levels of oxidative stress indicators SOD, GSH, LDH and MDA(P<0.05), inhibited the expression of inflammatory factors IL-6, IL-1β, iNOS and TNF-α(P<0.05) and up-regulated the expression of AMPK/Nrf-2/HO-1 signal pathway(P<0.05). CONCLUSION Notoginsenoside R1 improves myocardial injury in rats with coronary heart disease, inhibits cardiomyocyte apoptosis, oxidative stress, and inflammatory response, which is related to the activation of AMPK/Nrf-2/HO-1 signaling pathway.