目的：探讨红芪小分子提取物对骨髓间充质干细胞（ MSCs）体外增殖及染色体的影响。方法利用水提法结合超滤法提取红芪中小分子物质，并筛选其促进MSCs增殖的最佳浓度。设单独培养的MSCs为对照组，红芪最佳浓度（20 mg／L）培养72 h 后 MSCs为实验组，于倒置相差显微镜下观察各组细胞形态学的改变；采用MTT法检测各组细胞生长曲线；流式细胞术检测细胞周期；染色体显色与计数分析细胞染色体；West-ern印迹检测各组细胞基质金属蛋白酶（MMP）-2、MMP-9蛋白表达水平的变化。结果红芪浓度20 mg／L为最佳促进MSCs增殖浓度（ P＜0．05）。倒置相差显微镜下观察对照组细胞呈成纤维细胞样；红芪组细胞呈长梭形，形态类似于对照组。与对照组比较，红芪组细胞生长速度显著增快（P＜0．05）；细胞周期G0／G1期细胞减少，S期和G2／M期细胞增多，但差异无统计学意义（P＞0．05）；染色体及 MMP-2、MMP-9蛋白表达水平无异常改变（P＞0．05）。结论红芪小分子提取物20 mg／L体外培养MSCs 72 h后，对其增殖有明显促进作用，且维持其遗传物质染色体的稳定性，其作用机制可能与调控MSCs的MMP-2、MMP-9等相关蛋白表达有关。%Objective To study the effects of micromolecule-aqueous extracts of Hongqi on proliferation and chromosome of mesene-hymal stem cells ( MSCs) in vitro.Methods The water-boiling method combined with ultrafiltration were used to extract the micromolecule-aqueous substances of Hongqi and the best concentration was screened for proliferative effect on MSCs.MSCs cultured alone was control group,and MSCs cultured with the best concentration of Hongqi (20 mg/L) for 72 h was experimental groups.The morphological changes in the cells were observed by phase-contrast microscopy.MTT assay was used to measure the cells changes in growth.The cell cycle of MSCs was determined by flow cytometry.Chromosome was examined by colored and counted.The expressions of MMP-2,MMP-9 proteins were deter-mined by Western blot.Results 20 mg/L micromolecule-aqueous extracts of Hongqi was best concentrations to promote the growth of MSCs (P<0.05).The cell morphologies were observed by phase-contrast microscopy,cells of MSCs were fibroblast-like,cells of Hongqi were long fusiform,closed to MSCs cells in morphology.Compared with MSCs,cells of Hongqi grow were faster (P<0.05),the proportion of G0/G1 phase cells was reduced,while the proportion of S phase and G2/M phase were increased,but there was no statistical significance(P>0.05), the chromosome and the proteins expressions of MMP-2,MMP-9 did not change abnormally(P>0.05).Conclusions Micromolecule-aqueous extracts of Hongqi (20 mg/L) culturing MSCs for 72 h could improve proliferation of MSCs significantly,and preserve the genetic stability of MSCs'chromosome in vitro.The mechanism is related with the regulation of the expressions of MMP-2,MMP-9 proteins of MSCs.
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