玛诺系汤冠心宁浸膏对冠心病大鼠心肌损伤的保护作用及机制研究

Protective effects and mechanisms of Manuo Xitang Guanxinning extractum against myocardial injury in rats with coronary heart disease

关键词:  玛诺系汤冠心宁  冠心病  炎症  氧化应激

Key words:  Manuo Xitang Guanxinning  coronary heart disease  inflammation  oxidative stress

摘要:

目的 探究玛诺系汤冠心宁浸膏（GXNC）对冠心病（CHD）大鼠心肌损伤的保护作用及机制。方法 采用高脂饲料饲喂和垂体后叶素注射建立大鼠CHD模型，并实施GXNC干预。利用超高分辨率小动物超声影像系统，监测大鼠的心率（HR）、左心室舒张期内径（LVIDd）、左心室收缩期内径（LVIDs）、短轴缩短率（FS）和射血分数（EF）；利用全自动生化分析仪检测血清总胆固醇（TC）、三酰甘油（TG）、低密度脂蛋白胆固醇（LDL-C）、高密度脂蛋白胆固醇（HDL-C）、乳酸脱氢酶（LDH）、肌酸激酶同工酶-MB（CK-MB）和C反应蛋白（CRP）的水平；用ELISA试剂盒测定血清肌酸激酶（CK）、心肌肌钙蛋白I（cTnI）、丙二醛（MDA）、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）的水平；HE染色观察心肌组织的病理学变化；免疫组化技术检测心肌组织中IL-6、IL-1β和TNF-α的表达；分别测定心肌组织中TLR4、PI3K、AKT、mTOR mRNA及蛋白的相对表达。结果 与正常对照组相比，模型组大鼠心肌组织可见细胞变性、坏死，大量炎性细胞浸润。心功能指标：LVIDs、LVIDd，血脂指标：TC、TG和LDL?C，心肌损伤指标：LDH、CK、CK-MB、cTnΙ和CRP，氧化指标：MDA均发生极显著升高（P < 0.01）；促炎因子IL?6、IL-1β和TNF?α在心肌组织中呈阳性表达；而心功能指标：HR、EF和FS，氧化指标：SOD、GSH-Px，血脂指标：HDL-C，心肌组织中TLR4、PI3K、AKT、mTOR的mRNA及蛋白表达均显著降低（P < 0.05）；与模型组相比，GXNC高剂量组大鼠的心肌组织损伤、心功能指标、血脂指标、氧化指标、炎症因子及心肌组织中TLR4、PI3K、AKT、mTOR的mRNA及蛋白表达均发生显著改善（P < 0.05）。结论 玛诺系汤冠心宁浸膏能改善冠心病大鼠的心肌损伤和血脂水平，抑制其炎症和氧化应激反应，这与TLR4/PI3K/AKT/mTOR通路的激活有关。

Abstract:

Objective To investigate the protective effects and mechanisms of Manuo Xitang Guanxinning extractum against myocardial injury in rats with coronary heart disease (CHD). Methods Rat CHD model was established by high-fat diet feeding and pituitrin injection, and followed by Marrbio decoction Guanxinning extractum intervention. Heart rate (HR), left ventricular systolic diameter (LVIDs), left ventricular diastolic diameter (LVIDd), ejection fraction (EF) and short axis shortening rate (FS) of rats were measured by ultra-high resolution small animal ultrasound imaging system. Serum levels of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), lactate dehydrogenase (LDH), creatine kinase isoenzyme-MB (CK-MB) and C-reactive protein (CRP) were determined by automatic biochemical analyzer. Serum creatine kinase (CK), cardiac troponin I (cTnI), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) levels were determined by ELISA kits. Pathological changes of myocardial tissue in rats were observed with HE staining, and expressions of IL-6, IL-1β and TNF-α were detected by immunohistochemistry. Relative expressions of TLR4, PI3K, AKT and mTOR mRNA and proteins were determined respectively. Results Compared with the normal control group, myocardial tissue of rats in model group showed cell degeneration, necrosis and a large number of inflammatory cells infiltration. Cardiac function indicators: LVIDs and LVIDd, lipid indicators: TC, TG and LDL-C, myocardial injury indicators: LDH, CK, CK-MB, cTnΙ and CRP, and oxidative indicators: MDA all experienced highly significant elevations (P < 0.01).Pro-inflammatory factors IL 6, IL-1β and TNF-α positively expressed in myocardial tissue. Meanwhile, cardiac function indexes: HR, EF and FS, oxidative indexes: SOD, GSH-Px, lipid indexes: HDL-C, mRNA and protein expression of TLR4, PI3K, AKT and mTOR in myocardial tissue were significantly reduced (P < 0.05) Compared with model group, significant improvements in myocardial tissue injury, cardiac function index, lipid index, oxidative index, inflammatory factors and mRNA and protein expression of TLR4, PI3K, AKT and mTOR in myocardial tissue occurred in rats in the GXNC high-dose group (P < 0.05) Conclusion Manuo Xitang Guanxinning extractum can improve myocardial injury and blood lipids level of CHD rats, and inhibit inflammation and oxidative stress response, which is related to the activation of TLR4/PI3K/AKT/mTOR pathway.