中国组织工程研究Issue(7)：1243-1250,8.DOI:10.3969/j.issn.2095-4344.2013.07.019
过氧化氢对金属硫蛋白Ⅰ/Ⅱ敲除小鼠脾细胞的氧化应激***★
Hydrogen peroxide induced oxidative stress in the spleen of metallothionein knockout mice
摘要
Abstract
BACKGROUND: In the early stage of cerebral ischemia-reperfusion, there is a tremendous amount of inflammatory factor expression in the spleen. These inflammatory factor cause oxidative stress damage which leads to cel apoptosis in the spleen after cerebral ischemia-reperfusion. OBJECTIVE: To investigate the effect of exogenous hydrogen peroxide on spleen cel viability and the effect of N-acetyl-L-cytokine on protection of induced oxidative stress in the spleen of metal othionein-Ⅰ/Ⅱ knockout mice. METHODS: Metal othionein-Ⅰ/Ⅱ knockout mice spleen cellsuspension was prepared and treated with various concentrations of hydrogen peroxide (0.1, 0.2, 0.5, 1, 2 mmol/L) for 2 hours, and the cel viability was detected by MTT colorimetric method. Based on the cells and mitochondria level, hydrogen peroxide induced spleen cells were divided into six group: control group, N-acetyl-L-cytokine group, 0.5 mmol/L hydrogen peroxide group, 1 mmol/L hydrogen peroxide group, N-acetyl-L-cytokine+0.5 mmol/L hydrogen peroxide group, N-acetyl-L-cytokine+1 mmol/L hydrogen peroxide group. Then, the cel viability was detected by MTT colorimetric method, lactate dehydrogenate activity were assayed by microplate reader; mitochondrial permeability transition pore was evaluated by ultraviolet spectrophotometer after 2 hours. RESULTS AND CONCLUSION: With the increase of the concentration of hydrogen peroxide, the spleen cel viability was significantly decreased (P < 0.01), and 0.2, 0.5, 1, 2 mmol/L hydrogen peroxide groups had the greatest reduction. Compared with the control group, N-acetyl-L-cytokine could significantly increase spleen cel viability (P < 0.01), decrease lactate dehydrogenate activity (P < 0.01) and decrease the opening state of mitochondrial permeability transition pore (P < 0.01). Compared with 0.5 mmol/L hydrogen peroxide group and 1 mmol/L hydrogen peroxide group, N-acetyl-L-cytokine+0.5 mmol/L hydrogen peroxide group and N-acetyl-L-cytokine+1 mmol/L hydrogen peroxide group could also increase spleen cel viability (P < 0.01), decrease lactate dehydrogenate activity (P < 0.01) and decrease the opening state of mitochondrial permeability transition pore (P < 0.01), respectively. These findings suggest that with the increase of the concentration of hydrogen peroxide, the spleen cel viability is significantly decreased in a dose-dependent manner, especial y for 0.2, 0.5, 1, 2 mmol/L hydrogen peroxide; N-acetyl-L-cysteine can relieve the oxidative stress damage induced by hydrogen peroxide in metal othionein-Ⅰ/Ⅱ knockout mice spleen cells, through reducing the lactate dehydrogenase release and the opening state of mitochondrial permeability transition pore, and increasing spleen cel viability.关键词
组织构建/组织构建细胞学实验/金属硫蛋白/金属硫蛋白Ⅰ/Ⅱ基因敲除小鼠/脾/凋亡/N-乙酰-L-半胱氨酸/过氧化氢/氧化应激/线粒体通透性转换孔/脑缺血再灌注/乳酸脱氢酶/国家自然科学基金分类
医药卫生引用本文复制引用
和静,韦思宇,刘凤勇,叶静,沈炳玲,姚小梅..过氧化氢对金属硫蛋白Ⅰ/Ⅱ敲除小鼠脾细胞的氧化应激***★[J].中国组织工程研究,2013,(7):1243-1250,8.基金项目
国家自然科学基金(81273009)，天津市高等学校科技发展基金项目(20050107)，天津市应用基础及前沿技术研究计划(09JCYBJC11700)。Supported by：the National Natural Science Foundation of China, No.81273009* （81273009）
Tianjin High School Science &Technology Fund Planning Project, No.20050107* （）
Tianjin Science & Technology Council Grant of China, No.09JCYBJC11700* （）
