芒果苷对H_2O_2诱导的BRL细胞氧化损伤的保护作用

Protective effects of mangiferin on the models of BRL cell oxidative stress induced by H_2O_2

目的探讨芒果苷对过氧化氢诱导的正常大鼠肝BRL细胞氧化损伤的保护作用。方法选取正常大鼠肝细胞株BRL,通过过氧化氢诱导法制备细胞氧化损伤模型,实验设对照组、H_2O_2组、芒果苷组。HE染色、台盼蓝染色观察细胞形态变化和存活率变化;比色法检测细胞中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性和丙二醛(MDA)含量;Western blot检测细胞中Bcl-2、Bax和Caspase-3蛋白表达变化。结果对照组BRL细胞贴壁生长良好,排列紧密。H_2O_2诱导后细胞形态不规则,胞膜皱缩,贴壁能力下降;细胞存活率降低;抗氧化酶SOD、GSH-Px活性降低,MDA含量升高(P<0.05);Bax和Caspase-3蛋白表达升高(P<0.05)、Bcl-2蛋白表达降低(P<0.05)。与H_2O_2组相比,芒果苷可显著改善H_2O_2引起的细胞形态变化,提高细胞存活率;提高SOD、GSH-Px活性,降低MDA含量(P<0.05);抑制Bax和Caspase-3蛋白表达(P<0.05),上调Bcl-2蛋白表达(P<0.05)。结论芒果苷对BRL细胞氧化损伤具有保护作用,其作用机制可能与其提高细胞清除氧自由基能力和抑制细胞凋亡有关。

Objective To study the protective effect of mangiferin on BRL cell line injury induced by H2O2. Methods An oxidative stress model was established by treating BRL cell with H2O2. The BRL cell were divided into control group, H2O2 group, and mangiferin group. The morphology and activity changes of cell were observed by HE and trypan blue staining, respectively. The activity of superoxide dismutase （SOD）, glutathione peroxidase （GSH-Px）, and the content of malondialdehyde （MDA）were assayed by colorimetrie method. The Bcl-2, Bax and Caspase-3 protein expressions were detected by Western blot. Results BRL cells of the control group were adherent good growth, tightly arranged. After H2O2-induced, the shape of cells became irregular, accompanied by membrane shrinkage, and cell adherent ability decreasing. Compared with the control group, cell survival rate decreased, the activity of SOD and GSH-Px was lower （ P〈0.05 ） , and the content of MDA was higher （ P〈0.05 ） ; Caspase-3 and Bax protein expression levels were significantly increased （P〈0.05） , and Bel-2 protein expression levels were decreased （P〈0.05）. Mangiferin can significantly improve the H2O2-induced changes in cell morphology and cell survival rate, increase the activity of SOD and GSH-Px, and decreased the content of MDA（P〈0.05） ; Meanwhile, mangiferin can inhibit Bax, Caspase-3 protein expressions （P〈0.05）, and up-regulate Bcl-2 protein expression （P〈0.05）. Conclusion Mangiferin can protect the BRL cells against oxidative damage, its mechanism may be correlated with increase in oxygen free radicals removal ability and inhibition of apoptosis.