【摘要】：目的 本實(shí)驗(yàn)是在先前發(fā)現(xiàn)甘氨酸(Glycine)有心臟保護(hù)作用及在心肌細(xì)胞上發(fā)現(xiàn)有甘氨酸受體的基礎(chǔ)上，進(jìn)一步從自由基代謝、細(xì)胞內(nèi)鈣超載及細(xì)胞凋亡等方面探討甘氨酸受體在心臟保護(hù)中的作用，，并用甘氨酸受體阻斷法和消除細(xì)胞外氯離子法證明甘氨酸受體的性質(zhì)。 方法 1、利用原代培養(yǎng)的SD大鼠乳鼠心肌細(xì)胞建立心肌缺氧／復(fù)氧(anoxia／reoxygenation，A／R)模型，實(shí)驗(yàn)分為9組：(1)對(duì)照組；(2)甘氨酸組；(3)士的寧組；(4)消除細(xì)胞外氯離子+甘氨酸組；(5)A／R組；(6)(甘氨酸+A／R)組；(7)(士的寧+A／R)組；(8)(甘氨酸+士的寧+A／R)組；(9)(消除細(xì)胞外氯離子+甘氨酸+A／R)組。測(cè)定各組培養(yǎng)心肌細(xì)胞內(nèi)超氧化物歧化酶(SOD)、丙二醛(MDA)、一氧化氮(NO)的含量。 2、利用原代培養(yǎng)的SD大鼠乳鼠心肌細(xì)胞建立心肌缺氧／復(fù)氧(anoxia/reoxygenation，A／R)模型，實(shí)驗(yàn)分組同上。測(cè)定心肌細(xì)胞存活率，各組用Fluo—3／AM染色測(cè)鈣和Annexin V-FITC／PI雙染色測(cè)細(xì)胞凋亡率，流式細(xì)胞儀檢測(cè)。 結(jié)果 (1)A／R組SOD、NO含量低于對(duì)照組(P＜0.05)，而MDA含量高于對(duì)照組。A／R組用甘氨酸處理后SOD、NO含量升高，MDA含量降低，與A／R組比較差異有統(tǒng)計(jì)學(xué)意義。而A／R組用甘氨酸+士的寧處理后，甘氨酸的上述保護(hù)作用被士的寧阻斷，與A／R組比較差異無統(tǒng)計(jì)學(xué)意義。消除細(xì)胞外氯離子+甘氨酸+A／R時(shí)，甘氨酸的保護(hù)作用也被阻斷，與A／R組比較差異無統(tǒng)計(jì)學(xué)意義。 (2)A／R組心肌細(xì)胞內(nèi)游離鈣的濃度和細(xì)胞凋亡率高于對(duì)照組(P＜0.05)。A／R組用甘氨酸處理后心肌細(xì)胞內(nèi)游離鈣的濃度和細(xì)胞凋亡率降低，與A／R組
[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).
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R363

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