Objective To observe the protective effect of resveratrol(Res) against formaldehyde (FA)-induced oxidative stress in PC12 cells. Methods Rat adrenal pheochromocytoma monoclonal lines PC12 were cultured with 1640 in vitro and the cells in the exponential phase of growth to experiment were selected. The protective effect of Res on damages of PC12 induced by FA was investigated, the cells were divided into control, 80 μmol/L FA, solvent (2%Odms0), aminoguanidine (100 μmol/L) and Res (6.25, 12.5, 25, 50, 100 μmol/L) groups. Various concentrations Res were preincubated with cells for 2 h to detect indexes mentioned below. The viability of PC12 cells were detected by a colorimetric 3-[4, 5-dimethyl thiazol-2-yl]-2, 5-diphenyltetrazolium bromide ( MTT); the transudation content of lactate dehydrogenase ( LDH) was observed by colorimetry reaction; the degree of PC 12 cells injury was evaluated by measuring the content of malondialdehyde (MDA) in conditioned medium; the activity of superoxide dismutase (SOD) and the activity of glutathione (GSH-Px) in cells were measured by kits respectively. The level of nitric oxide(NO) was determined by nitrate reductase method; the activity of NOS and Inos were measured by biochemical approach. The cells in each group were collected, Western blot, PCR were used to detect the levels of Cytochrome C oxidase subunit II (CO II) protein and Mrna in PC1 2 cells. Apoptosis was measured by Hoechst 33258 fluorescence staining and Flow cytometry. Results FA induced cell membrane injury. When cells were preincubated with Res for 2 h, the levels of LDH, MDA, NO, NOS and Inos were reduced; the levels of SOD, GSH-Px were raised; the expressions of CO II protein and Mrna were enhanced, compared with those of 80 μmol/L FA group( P < 0. 01 or P < 0.05 ). Res alleviated FA induced oxidative damages in dose-dependent manner by Hoechst 33258 fluorescence staining and Flow cytometry, and apoptosis rate was decreased. Conclusions Res can protect PC12 cells from FA oxidatie stress injuries, improve cell survival rate. Res can enhance the expressions of CO II protein and Mrna, decrease the release of Cyt C from mitochondria, thereby Res can attenuate cell apoptosis.%目的 探讨白藜芦醇对甲醛诱导PC12细胞损伤干预的可能机制.方法 以甲醛损伤PC12细胞为氧化应激损伤模型,用不同浓度(6.25、12.5、25、50、100 μmol/L)的白藜芦醇预处理PC12细胞2 h后,加入终浓度为80 μmol/L甲醛作用24 h.并设氨基胍100 μmol/L为阳性对照组.采用四甲基偶氮噻唑蓝比色法检测细胞活力;分光光度法测定乳酸脱氢酶(LDH)漏出量;硫代巴比妥酸法测定MDA含量;黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)活性;二硫对二硝基苯甲酸(DTNB)比色法测定谷胱甘肽过氧化物酶(GSH-Px)活性;硝酸还原酶法测定一氧化氮(NO)含量、一氧化氮合酶(iNOS)活性;并收集各组细胞,检测细胞色素C氧化酶亚基Ⅱ(COⅡ)蛋白活性及mRNA的表达;Hoechst 33258荧光染色法和流式细胞技术检查细胞凋亡情况.结果 白藜芦醇能明显改善甲醛所致PC12细胞损伤,与甲醛处理组相比,细胞存活率随着白黎芦醇浓度的增加而加强,且100 μmol/L白藜芦醇组细胞存活率为(92.66±3.27)%,高于氨基胍阳性对照组[(82.18±2.06)%];在白黎芦醇6.25～100 μmol/L浓度范围内,SOD、GSH-Px活性、COⅡ蛋白及mRNA表达水平随着白藜芦醇浓度增加而逐渐升高;LDH漏出量、NO、MDA含量、iNOS活性随着处理浓度的增加而逐渐降低.结论 白藜芦醇对甲醛诱导PC12细胞损伤具有保护作用,在一定范围内呈剂量-效应关系;能有效改善甲醛所致PC12细胞的氧化损伤,提高细胞存活率;这种抗氧化活性可能与抑制甲醛/ROS、甲醛/NOS/NO氧化应激途径、减轻线粒体损伤有关.
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