氧化樟脑对氧化应激损伤后心肌细胞的保护作用

发布时间：2018-08-04 18:36

【摘要】：目的：通过氧化损伤标记物8-羟基脱氧鸟苷（8-OHDG）及细胞凋亡标记物半胱氨酸天冬氨酸蛋白酶-3（caspase-3）的检测，分析心肌缺血缺氧、心肌缺血再灌注、心肌细胞氧化应激的病理变化及相关机制，在此基础上研究氧化樟脑对氧化应激损伤后心肌细胞的保护作用。方法：采用H9c2大鼠心肌细胞复制缺氧模型，随机分为4组：正常组、氧化应激组、氧化应激+氧化樟脑A组、氧化应激+氧化樟脑B组。正常对照组：不作任何特殊处理；氧化应激组：将心肌细胞更换为无氧培养液，放入缺氧装置内，继续培养4h后，更换复氧液，再进行有氧培养4h，完成缺氧/复氧模型；氧化应激+氧化樟脑组：在心肌细胞缺氧/复氧完成后分别加入10ul/50ml（A组）、20ul/50ml（B组）浓度的氧化樟脑继续培养6h。通过噻唑蓝微量自动比色法（MTT）检测各组心肌细胞活力。通过免疫化学染色观察各组细胞氧化损伤标记物8-羟基脱氧鸟苷（8-OHDG）及细胞凋亡标记物半胱氨酸天冬氨酸蛋白酶-3（caspase-3）水平。统计学处理以Sigma Stat3.5软件进行统计学处理，应用t-Test或one-wayANOVA分析，结果以Mean±SD表示，P0.05为有统计学差异。结果： 1.氧化应激组H9c2心肌细胞继续培养6h后进行MTT检测。结果表明：氧化应激组吸光度值（A值）低于对照组（P0.05）。 2.氧化应激+氧化樟脑组H9c2心肌细胞继续培养6h后进行MTT检测。结果表明：氧化应激组吸光度值（A值）低于氧化应激+氧化樟脑组，氧化应激+氧化樟脑组吸光度值（A值）低于对照组（P0.05），氧化应激+氧化樟脑组较氧化应激组心肌细胞活力明显升高。氧化应激+氧化樟脑B组心肌细胞活力高于氧化应激+氧化樟脑A组。 3.8-OhdG表达在对照组最低，氧化应激组最高，而氧化应激+氧化樟脑组较氧化应激组明显降低，，氧化应激+氧化樟脑B组8-OhdG表达低于氧化应激+氧化樟脑A组。 4. caspase-3的蛋白表达在对照组最低，氧化应激组最高，而氧化应激+氧化樟脑组较氧化应激组明显降低，氧化应激+氧化樟脑B组的caspase-3蛋白表达低于氧化应激+氧化樟脑A组。结论：氧化樟脑对氧化应激损伤后心肌细胞具有保护作用，它可减轻氧化应激后心肌细胞的损伤，抑制氧化应激后心肌细胞的凋亡。
[Abstract]:Objective: to detect the oxidative damage marker 8-hydroxydeoxyguanosine (8-OHDG) and the apoptotic marker cysteine aspartate protease-3 (caspase-3) to analyze myocardial ischemia and hypoxia and myocardial ischemia-reperfusion. On the basis of pathological changes and related mechanisms of oxidative stress in cardiomyocytes, the protective effect of camphor on myocardial cells after oxidative stress was studied. Methods: hypoxia model was induced by H9c2 rat cardiomyocytes. The rats were randomly divided into 4 groups: normal group, oxidative stress oxidative camphor A group and oxidative stress oxidative camphor B group. Normal control group: without any special treatment, oxidative stress group: the cardiomyocytes were replaced with anaerobic culture medium, put into anoxic equipment, then cultured for 4 hours, then reoxygenated solution was changed, then aerobic culture was carried out for 4 hours to complete the hypoxia / reoxygenation model. Oxidative stress Oxycamphor group: after hypoxia / reoxygenation of cardiomyocytes were completed, 10ul/50ml (group A) 20 ulr / 50ml (group B) was added respectively to culture camphor for 6 h. The activity of myocardial cells in each group was detected by (MTT) with thiazolyl blue microcolorimetry. The levels of 8-hydroxydeoxyguanosine (8-OHDG) and cysteine aspartate protease -3 (caspase-3) were detected by immunocytochemical staining. Statistical processing was performed by Sigma Stat3.5 software, and t-Test or one-wayANOVA analysis was used. The result was that Mean 卤SD was used to show statistical difference (P0.05). Results: 1. In oxidative stress group, H9c2 cardiomyocytes were cultured for 6 h and MTT was detected. The results showed that the absorbance (A value) of oxidative stress group was lower than that of control group (P0.05). H9c2 cardiomyocytes in oxidative stress group were cultured for 6 h and MTT was detected. The results showed that the absorbance value (A value) of oxidative stress group was lower than that of oxidative stress oxidative camphor group. The absorbance (A value) of oxidative stress camphor group was lower than that of control group (P0.05). The activity of myocardial cells in oxidative stress camphor group was significantly higher than that in oxidative stress group. The activity of cardiomyocytes in oxidative stress camphor B group was higher than that in oxidative stress camphor A group. The expression of 3.8-OhdG was the lowest in the control group and the highest in the oxidative stress group, while the oxidative stress oxidative camphor group was significantly lower than that in the oxidative stress group. The expression of 8-OhdG in oxidative stress camphor B group was lower than that in oxidative stress camphor A group. 4. The protein expression of caspase-3 was the lowest in the control group and the highest in the oxidative stress group, while the oxidative stress oxidative camphor group was significantly lower than that in the oxidative stress group. The expression of caspase-3 protein in oxidative stress camphor B group was lower than that in oxidative stress camphor A group. Conclusion: Oxycamphor has protective effect on cardiomyocytes after oxidative stress. It can reduce the injury of cardiomyocytes after oxidative stress and inhibit the apoptosis of cardiomyocytes after oxidative stress.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

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