青心酮对H2O2诱导MC3T3-E1成骨细胞损伤的保护作用

The protective effect of 3,4-dihydroxyacetophenone on HO-induced osteoblastic MC3T3-E1 cells injury

目的探讨3,4-二羟基苯乙酮(青心酮)对过氧化氢(H2O2)诱导MC3T3-E1成骨细胞损伤的保护作用。方法建立青心酮预孵育MC3T3-E1成骨细胞后H2O2再损伤模型,采用MTT、流式细胞术和丙二醛(MDA)细胞盒分别观察青心酮(1、3、10μmol/L)预作用对H2O2诱导MC3T3-E1成骨细胞损伤的保护作用以及细胞内活性氧(ROS)和MDA水平的影响。采用免疫荧光实验检测青心酮对MC3T3-E1成骨细胞核因子E2相关因子2(Nrf2)入核的影响。使用小干扰RNA沉默Nrf2表达检测Nrf2介导青心酮对H2O2诱导MC3T3-E1成骨细胞损伤的保护作用。采用qRT-PCR和Western blot检测青心酮对MC3T3-E1成骨细胞血红素氧化酶1(HO-1)mRNA和蛋白水平的影响。结果青心酮预孵育MC3T3-E1成骨细胞后,能够增加H2O2损伤后细胞的存活率,降低细胞内ROS和MDA的水平(P<0.05)。青心酮可诱导MC3T3-E1成骨细胞中Nrf2进入细胞核,并通过激活Nrf2表达及提高HO-1的mRNA和蛋白表达对细胞起到保护作用(P<0.05)。结论青心酮通过激活Nrf2/HO-1通路对H2O2诱导MC3T3-E1成骨细胞损伤起保护作用。

Objective To study the protective effect of 3,4-dihydroxyacetophenone(DHAP)on H2O2-induced injury of osteoblastic MC3 T3-E1 cells.Methods The model of preincubation of MC3 T3-E1 cells with DHAP and H2O2 injury was established.The MTT,flow cytometry and malondialdehyde(MDA)cell cassette were used to investigate the protective effect of DHAP(1,3,10μmol/L)on MC3 T3-E1 cells induced by H2O2,the contents of intracellular ROS and MDA,respectively.The effect of DHAP on nuclear translocation of Nrf2 was detected by immunofluorescence assay.The protective effect of Nrf2 mediated-DHAP on H2O2-induced MC3 T3-E1 cells was detected by silencing Nrf2 expression using siRNA.qRT-PCR and Western blot were used to detect the effect of DHAP on HO-1 mRNA and protein levels,respectively.Results After pre-incubation of cells with DHAP,the protective effect of H2O2-induced cell injury was increased,and the levels of intracellular ROS and MDA were decreased(P<0.05).DHAP induced Nrf2 into the nucleus,and played a cytoprotective role through Nrf2 expression and increased HO-1 mRNA and protein expression(P<0.05).Conclusion DHAP has a protective effect on H2O2-induced injury of MC3 T3-E1 cells by activating Nrf2/HO-1 pathway.