欧亚旋覆花总黄酮通过抗氧化作用降低衰老大鼠骨髓间充质干细胞的凋亡

Inula Britannica flower total flavonoids reduces the apoptosis of aging bone marrow mesenchymal stem cells by anti-oxidation

目的探讨欧亚旋覆花总黄酮(IBFTF)对衰老大鼠骨髓间充质干细胞(BMSC)的改善效果及其机制。方法用D-半乳糖(D-galactose)诱导大鼠BMSC建立衰老细胞模型,采用(12.5、25、50)μg/m L IBFTF处理BMSC。CCK-8法检测细胞增殖,试剂盒测定过氧化氢酶(CAT)和超氧化物歧化酶(SOD)活性,丙二醛(MDA)和活性氧(ROS)的含量,流式细胞术检测细胞凋亡,Western blot法检测BAX、Bcl-2、裂解的胱天蛋白酶3(c-caspase-3)的蛋白水平。结果衰老组细胞的增殖率、SOD和CAT活性较对照组明显降低,而不同剂量的IBFTF促进D-Gal诱导的衰老细胞的增殖、增加SOD和CAT活性。衰老组的凋亡细胞比例、ROS含量、MDA水平、BAX/Bcl-2比值和c-caspase-3的蛋白表达量增加,而用不同浓度IBFTF治疗可显著降低衰老大鼠BMSC的凋亡、ROS的生成、MDA水平、BAX/Bcl2比值和c-caspase-3的蛋白水平。结论 IBFTF通过增强抗氧化作用减少衰老大鼠BMSC的凋亡。

Objective To investigate the beneficial effect of Inula Britannica flower total flavonoids（IBFTF） on aging bone mesenchymal stem cell（BMSC） and its potential mechanism.Methods The aging BMSCs were induced by D-galactose,and then treated with 12.5,25,50 μg/m L IBFTF.The cell viability was detected by CCK-8 assay.The activity of catalase（CAT） and superoxide dismutase（SOD）,the content of malondialdehyde（MDA） and reactive oxygen species（ROS）were measured by a commercial kit.The apoptosis was assessed by flow cytometry.The protein expressions of BAX,Bcl-2 and cleaved-caspase-3（c-caspase-3） were determined by Western blotting.Results The cell viability and the activity of SOD and CAT in the aging group decreased significantly compared with the normal group,whereas different concentrations of IBFTF promoted the cell viability,and simultaneously increased the activity of SOD and CAT.The apoptosis,the ROS production,the content of MDA,BAX/Bcl-2 ratio and the protein expression of c-caspase-3 in the aging group increased obviously compared with the normal group.However,the treatment of different concentrations of IBFTF reduced the apoptosis,the ROS production,the content of MDA,BAX/Bcl-2 ratio and the protein expression of c-caspase-3.Conclusion IBFTF can attenuate the apoptosis of aging BMSCs by anti-oxidation.