扶正化瘀方药物血清对人脐带间充质干细胞增殖、迁移及旁分泌能力的影响  

Effects of Fuzheng Huayu Decoction Contained Serum on Proliferation, Migration and Parapocrine Capacity of Human Umbilical Cord Mesenchymal Stem Cells

作　　者：栾玉玲 花东明 陈相波 王芳[1] 孔晓妮 冯煜[1] LUAN Yuling;HUA Dongming;CHEN Xiangbo;WANG Fang;KONG Xiaoni;FENG Yu(Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200120,China;Hangzhou Rongze Biotechnology Group Co.,Ltd,Hangzhou 311200,China)

机构地区：[1]上海中医药大学附属曙光医院,上海200120 [2]杭州荣泽生物科技集团有限公司,浙江杭州311200

出　　处：《中国中医药信息杂志》2021年第12期56-60,共5页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(81873582);上海中医药大学“研究生创新培养”专项科研项目(3489)。

摘　　要：目的观察扶正化瘀方药物血清对人脐带间充质干细胞(HUCMSCs)增殖、迁移及旁分泌能力的影响。方法贴壁法分离HUCMSCs,流式细胞术鉴定HUCMSCs表型并检测其分化能力;取第5代HUCMSCs,随机分为空白血清组及5%、10%、15%药物血清组,CCK-8法检测HUCMSCs增殖能力;克隆形成实验检测HUCMSCs增殖能力;划痕实验检测HUCMSCs迁移能力,测量划痕宽度并计算细胞迁移率。Transwell实验检测HUCMSCs迁移能力,统计迁移细胞数目。ELISA检测HUCMSCs旁分泌碱性成纤维细胞生长因子(bFGF)能力。结果分离培养的HUCMSCs高表达标志物CD90(99.8%)、CD105(97.1%)、CD73(99.9%)、CD44(100%),符合HUCMSCs表型特征并具有成脂/成骨分化功能。CCK-8结果显示,5%、10%、15%药物血清干预HUCMSCs后,均能促进HUCMSCs增殖(P<0.05)。克隆形成实验结果显示,与空白血清组比较,5%、10%、15%药物血清组HUCMSCs克隆形成数量增加。划痕实验和Transwell实验结果显示,与空白血清组比较,5%、10%、15%药物血清组干预后,HUCMSCs迁移率明显升高(P<0.01),且随药物血清浓度增加,细胞迁移率呈上升趋势。ELISA结果显示,与同一时点空白血清组比较,5%、10%、15%药物血清组HUCMSCs旁分泌b FGF能力明显增强(P<0.05)。结论扶正化瘀方药物血清对HUCMSCs增殖、迁移和旁分泌bFGF能力有明显促进作用。Objective To investigate the effects of Fuzheng Huayu Decoction contained serum on the proliferation,migration and paracocrine ability of human umbilical cord mesenchymal stem cells(HUCMSCs).Methods HUCMSCs were isolated by adherence method.The phenotype and the differentiation ability of HUCMSCs were examined by flow cytometry.The fifth generation of HUCMSCs were randomly divided into blank serum group,5%,10%and 15%drug-contained serum group,CCK-8 method was used to detect the proliferation ability of HUCMSCs.The proliferation ability of HUCMSCs was detected by clone formation experiment.The migration ability of HUCMSCs cells was detected by scratch adhesion test.Transwell assay was used to detect the migration ability of HUCMSCs.The ability of HUCMSCs to paracrine bFGF was detected by ELISA.Results The high expression markers of CD90(99.8%),CD105(97.1%),CD73(99.9%)and CD44(100%)in isolated and cultured HUCMSCs were consistent with the phenotypes of mesenchymal stem cells and had the ability of osteogenic and adipogenic differentiation.CCK-8 results showed that 5%,10% and 15%drug-contained serum could promote the proliferation of HUCMSCs(P<0.05).The clone formation experiment showed that,compared with the control group,the number of cell clone formation significantly increased in the 5%,10% and 15% drug-contained serum groups.Scratch adhesion test and Transwell assay results showed that,compared with the control group,the migration rate of HUCMSCs cells in 5%,10% and 15% drug-contained serum groups significantly increased(P<0.01),and the cell migration rate showed increased trend with the increased drug-contained serum.The ELISA result showed that,compared with the control group at the same period,the ability of HUCMSCs to paracrine bFGF in 5%,10% and 15% drug-contained serum group increased(P<0.05).Conclusion Fuzheng Huayu Decoction contained serum can significantly promote the abilities of proliferation,migration and paracrine bFGF of HUCMSCs.

关 键 词：扶正化瘀方 人脐带间充质干细胞 细胞增殖 细胞迁移 旁分泌 

分 类 号：R285.5[医药卫生—中药学]