冠心宁片对大鼠心肌缺血再灌注损伤的心肌保护作用及机制研究

发布时间：2017-12-27 22:22

本文关键词：冠心宁片对大鼠心肌缺血再灌注损伤的心肌保护作用及机制研究　出处：《浙江中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的观察冠心宁片(GXNT)对心肌缺血再灌注损伤(MI/RI)大鼠的心肌保护作用,并探讨其作用机制。方法SD大鼠70只,随机分为7组,即伪手术组、MI/RI组、GXNT 75mg/kg,150mg/kg,300mg/kg,600mg/kg 剂量组和 300mg/kg 复方丹参片(DST)组,每组10只。所有大鼠均预防性经口灌胃给药7天后,采用结扎大鼠左冠状动脉前降支法,建立MI/RI大鼠模型。期间记录大鼠心电图,分析心电图J点和心率变异性(HRV)指标;用伊文思蓝(EB)和2,3,5-氯化三苯基四氮唑蓝(TTC)双重染色心肌并分析心肌梗死面积,测定血浆天门冬氨酸氨基转氨酶(AST)活性、乳酸脱氢酶(LDH)活性、肌酸激酶(CK)及其同工酶(CK-MB)活性、超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量和一氧化氮(NO)水平,同时取部分心肌进行组织病理HE染色,观察心肌病理改变情况;另取部分心肌组织进行组织匀浆,测定心肌组织中SOD活性、MDA含量、NO水平、髓过氧化物酶(MPO)活性和总抗氧化能力(T-AOC)水平的变化。结果与伪手术组比,MI/RI模型大鼠冠脉结扎后缺血期心电图J点明显抬高(P0.01),再灌注后J点虽均有所降低,但仍显著高于伪手术组(P0.01),RR间期、SDNN、RMSSD、TP和HFnu均出现明显降低(P0.05,P0.01),LFnu和LF/HF比值显著升高(P0.01),同时心肌缺血范围(ARR/WH和ARR/LV)和梗死范围(INF/WH、INF/LV和INF/ARR)均显著增加(P0.01),血浆AST、LDH、CK和CK-MB活性均明显升高(P0.01),血浆和心肌组织中SOD活性、T-AOC活性和NO活性均明显降低(P0.05,P0.01),MDA含量和MPO活性均明显升高(P0.05,P0.01),心肌纤维肿胀、紊乱、呈波浪样改变、部分心肌纤维断裂、细胞核有破裂、溶解消失现象,心肌间质有炎性细胞浸润,并可见局灶性坏死和出血,提示MI/RI模型大鼠出现明显的心肌损伤,心肌自由基生成增多,存在明显的氧化应激和炎症反应,导致心脏自主神经功能紊乱的状态。给予冠心宁片和复方丹参片后能明显降低再灌注期J点、∑-J值和J点平均值(P0.05,P0.01),显著降低心肌缺血范围和梗死面积(P0.05,P0.01),能明显升高SDNN、RMSSD、TP和HFnu以及降低LFnu和LF/HF比值(P0.05,P0.01),同时能抑制血浆AST、LDH、CK和CK-MB活性的释放(P0.05,P0.01),提高血浆SOD活性和NO水平(P0.05,P0.01),并能降低MDA含量(P0.05,P0.01),同时能显著提高心肌组织SOD活性、T-AOC活性、NO水平和降低心肌组织MDA含量和MPO活性(P0.05,P0.01),并能改善心肌病理组织结构。结论冠心宁片能通过抗氧化和抑制炎症反应,改善心脏自主神经调控功能,从而减轻MI/RI大鼠心肌梗死,具有抗MI/RI的作用。
[Abstract]:Objective To observe the effect of Guanxinning tablet (GXNT) on myocardial ischemic reperfusion injury (MI/RI) myocardial protective effect in rats, and to explore its mechanism of action. Methods 70 SD rats were randomly divided into 7 groups, namely, the pseudo operation group, MI/RI group, GXNT 75mg/kg, 150mg/kg, 300mg/kg, 600mg/kg dose group and 300mg/kg Compound Salvia Tablet (DST) group, 10 rats in each group. The rat model of MI/RI was established by ligation of the left anterior descending branch of the coronary artery in all rats after 7 days of prophylactic administration of the oral administration. During the recording of the electrocardiogram of rats, J analysis of ECG and heart rate variability index (HRV); Evans blue (EB) and 2,3,5- chloride three phenyl tetrazolium (TTC) double staining and analysis of myocardial infarct size, determination of plasma aspartate aminotransferase (AST) activity, lactate dehydrogenase (LDH) activity, creatine kinase (CK) and its isoenzyme (CK-MB) activity, superoxide dismutase (SOD) activity, malondialdehyde (MDA) content and nitric oxide (NO) level, at the same time, part of Myocardium Pathological HE staining, observe the changes of cardiomyopathy; another part of the myocardial tissue homogenate in myocardial tissue by SOD the activity, MDA content, NO level, myeloperoxidase (MPO) activity and total antioxidant capacity (T-AOC) level change. The pseudo operation group, MI/RI model of coronary artery ligation in rats after ischemia ECG J point elevation (P0.01), J after reperfusion is decreased, but still higher than the sham operation group (P0.01), SDNN, RMSSD, RR interval, TP and HFnu were significantly reduced (P0.05, P0.01), LFnu and LF/HF were significantly increased (P0.01), and the extent of myocardial ischemia (ARR/WH and ARR/LV) and infarct size (INF/WH, INF/LV and INF/ARR) were significantly increased (P0.01), AST, LDH, CK and plasma CK-MB activity were significantly increased (P0.01), plasma and myocardial tissue SOD activity, T-AOC activity and the activity of NO was significantly decreased (P0.05, P0.01), MDA content and MPO activity were significantly increased (P0.05, P0.01), myocardial fiber swelling and disorder, a wavy, fracture, part of myocardial fiber rupture, nuclear dissolution phenomenon, myocardial interstitial inflammatory cell infiltration, and visible focal Sexual necrosis and hemorrhage showed that MI/RI rats showed obvious myocardial injury, increased free radicals and obvious oxidative stress and inflammatory reaction, leading to cardiac autonomic dysfunction. Given Guanxinning tablet and Compound Salvia Tablet can significantly reduce the reperfusion period of J, -J and J values of sigma point average (P0.05, P0.01), significantly reduced the extent of myocardial ischemia and infarction area (P0.05, P0.01), can significantly increase SDNN, RMSSD, TP and HFnu and the decrease of LFnu and the ratio of LF/HF (P0.05, P0.01) at the same time, can inhibit the plasma AST, LDH, CK and CK-MB activity (P0.05, P0.01) release, improve the activity of plasma SOD and NO levels (P0.05, P0.01), and can reduce the content of MDA (P0.05, P0.01), at the same time can significantly improve the myocardial SOD activity, T-AOC activity, NO level and decrease the content of MDA and the activity of MPO in myocardial tissue (P0.05, P0.01), and can improve the myocardial pathological structure. Conclusion Guanxinning tablet can inhibit inflammatory reaction by anti-oxidation and, improve the regulation of autonomic nerve function, reduce myocardial infarction in MI/RI rats, with anti MI/RI effect.
【学位授予单位】：浙江中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

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