甘氨酸受体在心肌细胞缺氧复氧损伤中的作用机制研究

发布时间：2018-11-20 19:54

【摘要】：目的本实验是在先前发现甘氨酸(Glycine)有心脏保护作用及在心肌细胞上发现有甘氨酸受体的基础上，进一步从自由基代谢、细胞内钙超载及细胞凋亡等方面探讨甘氨酸受体在心脏保护中的作用，，并用甘氨酸受体阻断法和消除细胞外氯离子法证明甘氨酸受体的性质。方法 1、利用原代培养的SD大鼠乳鼠心肌细胞建立心肌缺氧／复氧(anoxia／reoxygenation，A／R)模型，实验分为9组：(1)对照组；(2)甘氨酸组；(3)士的宁组；(4)消除细胞外氯离子+甘氨酸组；(5)A／R组；(6)(甘氨酸+A／R)组；(7)(士的宁+A／R)组；(8)(甘氨酸+士的宁+A／R)组；(9)(消除细胞外氯离子+甘氨酸+A／R)组。测定各组培养心肌细胞内超氧化物歧化酶(SOD)、丙二醛(MDA)、一氧化氮(NO)的含量。 2、利用原代培养的SD大鼠乳鼠心肌细胞建立心肌缺氧／复氧(anoxia/reoxygenation，A／R)模型，实验分组同上。测定心肌细胞存活率，各组用Fluo—3／AM染色测钙和Annexin V-FITC／PI双染色测细胞凋亡率，流式细胞仪检测。结果 (1)A／R组SOD、NO含量低于对照组(P＜0.05)，而MDA含量高于对照组。A／R组用甘氨酸处理后SOD、NO含量升高，MDA含量降低，与A／R组比较差异有统计学意义。而A／R组用甘氨酸+士的宁处理后，甘氨酸的上述保护作用被士的宁阻断，与A／R组比较差异无统计学意义。消除细胞外氯离子+甘氨酸+A／R时，甘氨酸的保护作用也被阻断，与A／R组比较差异无统计学意义。 (2)A／R组心肌细胞内游离钙的浓度和细胞凋亡率高于对照组(P＜0.05)。A／R组用甘氨酸处理后心肌细胞内游离钙的浓度和细胞凋亡率降低，与A／R组
[Abstract]:Objective to study the effects of glycine (Glycine) on cardiac protection and glycine receptor in cardiomyocytes. The role of glycine receptor in cardiac protection was investigated in the aspects of intracellular calcium overload and apoptosis. The properties of glycine receptor were proved by blocking glycine receptor and eliminating extracellular chloride ion. Methods 1. Myocardial hypoxia / reoxygenation (anoxia/reoxygenation,A/R) model was established by primary cultured neonatal SD rat cardiomyocytes. The model was divided into 9 groups: (1) control group, (2) glycine group, and (2) glycine group. (3) strychnine group; (4) elimination of extracellular chloride ion glycine group; (5) A / R group; (6) glycine A / R group; (7) strychnine A / R group; (8) glycine strychnine A / R group; (9) (elimination of extracellular chloride ion glycine A / R) group. The content of superoxide dismutase (SOD),) malondialdehyde (MDA),) nitric oxide (NO) in cultured cardiomyocytes was measured. 2Myocardial hypoxia / reoxygenation (anoxia/reoxygenation,A/R) model was established by primary cultured neonatal SD rat cardiomyocytes. The survival rate of cardiomyocytes was measured. The apoptosis rate was measured by Fluo-3/AM staining and Annexin V-FITC/PI double staining. Flow cytometry was used to detect the apoptosis rate. Results (1) the content of SOD,NO in group A / R was lower than that in control group (P < 0. 05), while the content of MDA in group A / R was higher than that in control group. The content of SOD,NO increased and the content of MDA decreased after treatment with glycine in group A / R (P < 0. 05). The difference was statistically significant compared with the A / R group. The protective effect of glycine was blocked by strychnine in group A / R, and there was no significant difference between group A / R and group A / R. The protective effect of glycine was also blocked when the extracellular chloride ion glycine A / R was eliminated, and there was no significant difference compared with the A / R group. (2) the concentration of intracellular free calcium and the rate of apoptosis in the group A / R were higher than those in the control group (P < 0.05), the concentration of free calcium and the rate of apoptosis in the group of A / R were lower than those in the group of A / R (P < 0.05).
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363

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