不同频次调髓中药对人脐带间充质干细胞的作用

Effect of different frequencies of marrow-modulating herbs on human umbilical cord-derived mesenchymal stem cells

背景:间充质干细胞(MSCs)作用的微环境需要改善,调髓中药有不同频次,并且对MSCs具有调控作用。目的:比较不同频次调髓中药对人脐带间充质干细胞(huc-MSCs)的作用。方法:将肉桂、肉苁蓉、当归、川芎、白术和何首乌稀释成不同浓度,对huc-MSCs进行干预,采用CCK-8法检测不同浓度、不同频次调髓中药对huc-MSCs活力的影响;细胞划痕实验和Transwell迁移实验检测肉苁蓉和川芎对huc-MSCs迁移能力的影响;qPCR检测肉苁蓉和川芎对huc-MSCs CDC25A、CCNA2、E2F1、E2F2、MMP-2、TWIST1、VITMENTIN、PCNA mRNA表达的影响。结果:CCK-8结果显示高频调髓中药肉桂、肉苁蓉、当归能显著增强huc-MSCs的细胞活力,而低频调髓中药川芎、白术、何首乌对huc-MSCs的促增殖作用随着浓度的增加而降低;细胞划痕实验和Transwell迁移实验结果显示肉苁蓉可以促进huc-MSCs迁移(P<0.05或0.01),而川芎抑制huc-MSCs迁移(P<0.05或0.01);qPCR结果显示肉苁蓉能促进huc-MSCs CDC25A、CCNA2、E2F1、E2F2、MMP-2、TWIST1、VITMENTIN、PCNA mRNA表达(P<0.05或0.01),川芎抑制huc-MSCs E2F1、E2F2、MMP-2 mRNA表达(P<0.05或0.01)。结论:高频调髓中药在体外可以促进huc-MSCs增殖和迁移,而且其促增殖作用大于低频调髓中药。低频调髓中药抑制huc-MSCs迁移。其作用可能与细胞周期进程、细胞迁移、细胞增殖相关基因有关。

Background:The microenvironment of mesenchymal stem cells(MSCs)needs to be improved.Marrowmodulating herbs have different frequencies and modulating effects on MSCs.Objective:To compare the effects of different frequencies of marrow-regulating herbs on human umbilical cord-derived MSCs(huc-MSCs).Methods:Cinnamon,Cistanche,Angelica,Chuanxiong,Atractylodes and He Shou Wu were diluted into different concentrations to interfere with huc-MSCs.The CCK-8 method was used to detect the effects of different concentrations and frequencies of marrow-modulating herbs on the viability of huc-MSCs.Cell wound scratch assay and Transwell assay were used to detect the effects of Cistanche and Chuanxiong on the migration ability of huc-MSCs.qPCR was performed to detect the effects of Cistanche and Chuanxiong on the mRNA expression of huc-MSCs CDC25A,CCNA2,E2F1,E2F2,MMP-2,TWIST1,VITMENTIN,and PCNA.Results:CCK-8 results showed that high-frequency Cinnamon,Cistanches and Angelicae significantly stimulated the cell viability of huc-MSCs;low-frequency Chuanxiong,Atractylodes and He Shou Wu promoted proliferation of huc-MSCs,but the effect decreased with increasing drug concentrations.Cell scratching assay and Transwell assay showed that Cistanches promoted the migration of huc-MSCs(P<0.05,0.01),while Chuanxiong inhibited the migration of huc-MSCs(P<0.05,0.01).The qPCR results showed that Cistanche upregulated CDC25A,CCNA2,E2F1,E2F2,MMP-2,TWIST1,VITMENTIN and PCNA mRNA expression of huc-MSCs(P<0.05,0.01),while Chuanxiong inhibited E2F1,E2F2 and MMP-2 mRNA expression of hucMSCs(P<0.05,0.01).Conclusions:High-frequency marrow-modulating herbs can promote the proliferation and migration of huc-MSCs in vitro,and their pro-proliferative function was higher than those of low-frequency marrow-modulating herbs.In addition,low-frequency marrow-modulating herbs inhibit the migration of huc-MSCs,which may be related to the regulation of cell cycle,cell migration,and cell proliferation-related genes.