肺金生方对骨髓间充质干细胞的体外抗窜扰作用  

Anti-tampering effect of Feijinsheng Formula on bone marrow mesenchymal stem cells in vitro

作　　者：陈滨海 张雅丽 陈卫建 楼招欢[2] 李晓娟[3] 高文仓 张光霁[2] CHEN Bin-hai;ZHANG Ya-li;CHEN Wei-jian;LOU Zhao-huan;LI Xiao-juan;GAO Wen-cang;ZHANG Guang-ji(The Second Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310005,China;Zhejiang Chinese Medical University,Hangzhou 310053,China;The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310006,China)

机构地区：[1]浙江中医药大学附属第二医院,杭州310005 [2]浙江中医药大学,杭州310053 [3]浙江中医药大学附属第一医院,杭州310006

出　　处：《中华中医药杂志》2022年第6期3515-3518,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81273904);浙江省自然科学基金项目(No.LY19H270002);浙江省中医药科技计划项目(No.2019ZZ011,No.2020ZB116)。

摘　　要：目的:探讨肺金生方在体外对骨髓间充质干细胞(BMSCs)的抗窜扰作用。方法:利用Transwell小室,建立BMSCs与A549细胞共培养模型,以肺金生方冻干粉作用于BMSCs后,普通倒置显微镜观察细胞的形态,CCK8法检测细胞增殖情况,流式细胞术法分析细胞α-SMA表达阳性率,ELISA法检测成纤维细胞特异蛋白-1(FSP-1)、波形蛋白(vimentin)及α-平滑肌动蛋白(α-SMA)分子的表达,进一步用Western blot检测细胞中FSP-1、vimentin及α-SMA蛋白表达水平。结果:梯度浓度的肺金生方冻干粉作用于BMSCs 24 h后,1 mg/mL肺金生方可显著抑制BMSCs的活力,其细胞存活率较对照组(0μg/mL)显著降低(P<0.01);细胞形态由规则的梭形、椭圆形转变为不规则形,细胞数量减少,聚集情况减少;流式细胞仪检测BMSCs与A549细胞共培养后α-SMA表达阳性率上升(P<0.05)。与空白组比较,肺金生方组显著下调了FSP-1、vimentin及α-SMA表达(P<0.05);Western blot法证实,1000μg/mL肺金生方下调了FSP-1、vimentin及α-SMA的蛋白表达(P<0.05,P<0.01),一定范围内呈浓度梯度依赖性。结论:肺金生方对BMSCs具有一定的抗窜扰作用。Objective:To investigate the anti-tampering effect of Feijinsheng Formula on bone marrow mesenchymal stem cells(BMSCs)in vitro.Methods:Using Transwell chambers,BMSCs and A549 cell co-culture model was established,Transwell chamber was used to establish the co-culture model of BMSCs and A549 cells.After treating BMSCs with Feijinsheng formula freeze-dried powder,the cell morphology was observed under inverted microscope,cell proliferation was detected by CCK8 method,and the positive rate ofα-SMA expression was analyzed by flow cytometry.The expressions of fibroblast specific protein-1(FSP-1),vimentin andα-smooth actin(α-SMA)were detected by ELISA,and the expression levels of FSP-1,vimentin andα-SMA were further detected by Western blot.Results:After treated with gradient concentration of Feijinsheng Formula lyophilized powder for 24 h,1 mg/mL Feijinsheng Formula significantly inhibited the activity of BMSCs,and the cell survival rate was significantly lower than that of the control group(0μg/mL)(P<0.05).The cell morphology changed from regular spindle shape and ellipse shape to irregular shape,the number of cells decreased and the aggregation of cells decreased.The positive rate ofα-SMA expression was increased by flow cytometry after co-culture of BMSCs and A549 cells(P<0.05).ELISA detected that Feijinsheng Formula group down-regulated the expressions of FSP-1,vimentin andα-SMA,and there was a difference compared with blank group(P<0.05).Western blot analysis confirmed that 1000μg/mL Feijinsheng Formula down-regulated the expressions of FSP-1,vimentin andα-SMA in a concentration-gradient dependent manner within a certain range,with statistically significant differences(P<0.05,P<0.01).Conclusion:Feijinsheng Formula has a certain anti-tampering effect on BMSCs.

关 键 词：肺金生方 肺癌 骨髓间充质干细胞 A549细胞 抗窜扰 体外研究 

分 类 号：R285[医药卫生—中药学]