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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 effect and mechanism of total flavonoids of Inula vulgaris (IBFTF) on bone marrow mesenchymal stem cells (BMSC) in aging rats. Methods The senescent cell model was induced by D-galactose in rat BMSCs. BMSCs were treated with (12.5, 25, 50 μg / mL) IBFTF. The cell proliferation was detected by CCK-8 assay. The activity of catalase (CAT) and superoxide dismutase (SOD), malondialdehyde (MDA) and reactive oxygen species (ROS) The protein level of BAX, Bcl-2 and cleaved caspase-3 were detected by Western blot. Results The proliferation rate of cells in aging group and the activity of SOD and CAT were significantly lower than those in control group. However, different doses of IBFTF promoted the proliferation of senescent cells induced by D-Gal and increased the activity of SOD and CAT. The percentage of apoptotic cells, ROS content, MDA level, BAX / Bcl-2 ratio and c-caspase-3 protein expression in aging group were increased, while treatment with different concentrations of IBFTF could significantly reduce the apoptosis of BMSC MDA level, BAX / Bcl2 ratio and c-caspase-3 protein level. Conclusion IBFTF can reduce the apoptosis of BMSCs in aging rats by enhancing the anti-oxidative effect.