槲皮素调控Nrf2/HO-1信号通路抗氧化应激的机制研究

Quercetin Regulates Nrf2/HO-1 Signaling Pathway Against Oxidative Stress

目的:研究氧化应激状态下槲皮素对MC3T3-E1细胞的保护作用及分子机制。方法:使用200μmol/L过氧化氢干预细胞,构建氧化应激损伤模型。细胞增殖实验检测槲皮素对MC3T3-E1细胞增殖的影响;MDA、SOD试剂盒检测细胞中MDA、SOD含量;ALP染色、ARS染色检测槲皮素对MC3T3-E1细胞成骨分化的影响;qRT-PCR、Western blot检测细胞内Runx2、Osterix、Bax、Bcl-2、Caspase3、Nrf2、HO-1等基因和蛋白的表达情况。结果:CCK-8实验结果显示,槲皮素能够改善过氧化氢对MC3T3-E1细胞增殖抑制的影响(P<0.05)。过氧化氢显著提高了细胞中MDA含量,降低了SOD活性(P<0.05),然而,槲皮素处理后提高了MC3T3-E1细胞的抗氧化能力。ALP染色和ARS染色结果显示槲皮素能够挽救过氧化氢造成的MC3T3-E1细胞成骨分化能力减弱。qRT-PCR结果显示槲皮素干预后提高了Osterix、Runx2和Bcl-2的表达,降低了Bax、Caspase3的基因表达(P<0.05)。Western blot结果显示槲皮素提高了Nrf2、HO-1、Osterix、Runx2和Bcl-2的表达,降低了Bax、Caspase3的蛋白表达(P<0.05)。结论:槲皮素能够通过调控Nrf2/HO-1信号通路减轻过氧化氢诱导的氧化应激损伤,挽救MC3T3-E1细胞的增殖和成骨分化能力。

Objective:This study set out to investigate the protective effect of quercetin on MC3T3-E1 cells under oxidative stress and the molecular mechanism.Methods:Cells were intervened with 200μmol/L hydrogen peroxide to construct a model of oxidative stress injury.Cell proliferation assay was performed to detect the effects of quercetin on the proliferation of MC3T3-E1 cells;MDA and SOD assay kits were used to detect the contents of MDA and SOD in cells;ALP staining and alizarin red S staining were used to detect the effects of quercetin on osteogenic differentiation of MC3T3-E1 cells;qRT-PCR and Western blot were used to determine the intracellular expression of Runx2,Osterix,Bax,Bcl-2,Caspase3,Nrf2,HO-1.Results:The results of CCK-8 assay showed that quercetin ameliorated the effect of hydrogen peroxide on proliferation inhibition of MC3T3-E1 cells(P<0.05).Hydrogen peroxide significantly increased the MDA content and decreased the SOD activity in the cells(P<0.05);however,quercetin treatment improved the antioxidant capacity of MC3T3-E1 cells.ALP staining and alizarin red S staining results showed that quercetin was able to rescue the diminished osteogenic differentiation of MC3T3-E1 cells caused by hydrogen peroxide.qRT-PCR results showed that quercetin intervention increased the expression of Osterix,Runx2 and Bcl-2 and decreased the gene expression of Bax and Caspase3(P<0.05).western blot results showed that quercetin increased the expression of Nrf2,HO-1,Osterix,Runx2 and Bcl-2 and decreased the protein expression of Bax and Caspase3(P<0.05).Conclusion:Quercetin attenuates hydrogen peroxide-induced oxidative stress injury and rescues the proliferation and osteogenic differentiation ability of MC3T3-E1 cells by regulating the Nrf2/HO-1 signaling pathway.