维生素C增强衰老个体来源的骨髓间充质干细胞的增殖能力

Vitamin C promotes in vitro proliferation of bone marrow mesenchymal stem cells derived from aging mice

目的观察维生素C(Vc)能否通过升高端粒酶活性促进衰老个体来源骨髓间充质干细胞(BMMSCs)的增殖能力。方法以骨特异性早衰小鼠(SAMP6)为实验组,不发生早衰的R品系小鼠(SAMR1)为对照组,采用显微CT扫描技术验证SAMP6小鼠的骨衰老表型。分离培养两组小鼠的BMMSCs,其中SAMP6小鼠来源的BMMSCs分别应用不同浓度的Vc进行处理。采用MTT法检测BMMSCs的增殖能力,绘制生长曲线,使用端粒酶试剂盒检测端粒酶活性,并用PCR和Western blot方法检测端粒酶逆转录酶(TERT)的表达水平。结果 SAMP6小鼠具有骨衰老表型,SAMP6小鼠来源的BMMSCs的增殖能力和端粒酶活性均低于SAMR1小鼠来源的BMMSCs,差异均有统计学意义(P<0.05)。加入Vc后,在一定浓度范围内SAMP6小鼠来源的BMMSCs增殖能力随Vc浓度的升高而显著改善(P<0.05),同时其端粒酶活性和TERT表达水平有所提高(P<0.05),与增殖能力具有相关性。其中,Vc发挥促进作用的最适浓度为100μg/m L,在1000μg/m L时有抑制作用。结论维生素C能够增强衰老个体来源BMMSCs的增殖能力,且这种作用可能是通过升高端粒酶活性实现的。

Objective To investigate whether vitamin C can promote the proliferation ability of bone marrow mesenchymal stem cells（BMMSCs） derived from aging mice. Methods The senescence- accelerated mouse prone 6（SAMP6） mice and senescence- accelerated mouse resistant 1（SAMR1） mice were used as the test group and the control group, respectively, and the SAMP6 mice were examined by micro-CT to verify the senescent phenotype. BMMSCs were harvested from the two mouse lines and cultured in vitro, and the cells from SAMP6 mice were subjected to treatment with different concentrations of vitamin C. The proliferation ability of the cells from the two mouse lines was tested using MTT assay and growth curves, and Telo TAGGG Telomerase PCR ELISA was used to measure the telomerase activity; PCR and Western blotting were performed to detect the expression level of telomerase reverse transcriptase（TERT） in the cells. Results The SAMP6 mice displayed a bone senescent phenotype. The proliferation ability of BMMSCs derived from SAMP6 mice and their telomerase activity were significantly lower than those derived from SAMR1 mice（P〈0.05）. Vitamin C treatment significantly enhanced the proliferation ability of BMMSCs derived from SAMP6 mice in a dose- dependent manner（P〈0.05） and increased telomerase activity and TERT expression in the cells（P〈0.05）. At the concentration of 100 μg/m L, vitamin C produced the strongest effect in promoting the proliferation of BMMSCs from SAMP6 mice, while at the concentration of 1000 μg/ml, growth suppression occurred in the cells. Conclusion Vitamin C can promote the proliferation of BMMSCs from aging mice possibly by increasing the cellular telomerase activity.