月腺大戟素A干预CXCR4表达对间充质干细胞向心肌梗死区迁移的影响  

Effects of Ebracteolatain A on Migration of MSCs to Infarcted Region of Heart by Regulating CXCR4 Expression

作　　者：赵玥 王莹[2] 赵亮 李兰英 王奕[1] 牌铭欣 张贵振 王强利[1] ZHAO Yue;WANG Ying;ZHAO Liang;LI Lanying;WANG Yi;PAI Mingxin;ZHANG Guizhen;WANG Qiangli(School of Basic Medical Sciences,Shanghai University of TCM,Shanghai 201203,China;School of Pharmacy,Shanghai University of TCM,Shanghai 201203,China;Shanghai Baoshan Luodian Hospital,Shanghai 201908,China;Fourth People’s Hospital Affiliated to Tongji University,Shanghai 200081,China)

机构地区：[1]上海中医药大学基础医学院,上海201203 [2]上海中医药大学中药学院,上海201203 [3]上海宝山区罗店医院,上海201908 [4]同济大学附属第四人民医院,上海200081

出　　处：《中国中医药信息杂志》2020年第7期65-69,共5页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81873366、81673729);国家自然科学基金青年项目(81703763);上海中医药大学基础医学院攀登计划(GY010101)。

摘　　要：目的观察月腺大戟素A对CXC趋化因子受体4(CXCR4)表达的影响,探讨其调节间充质干细胞(MSCs)向心肌梗死区迁移的作用机制。方法采用双荧光素酶报告基因检测技术评价月腺大戟素A增强CXCR4启动子活性的量效关系,采用CCK-8试剂盒检测月腺大戟素A干预MSCs细胞活力,采用Transwell实验检测月腺大戟素A干预MSCs迁移能力,采用Western blot检测月腺大戟素A干预MSCs的CXCR4表达,采用小干扰RNA(siRNA)技术检测月腺大戟素A干预CXCR4表达MSCs的迁移。建立大鼠心肌梗死模型,在体评估月腺大戟素A干预绿色荧光蛋白(GFP)标记MSCs的迁移,同时采用Western blot检测月腺大戟素A干预MSCs中蛋白激酶B(Akt)活性和基质金属蛋白酶2(MMP2)的表达。结果月腺大戟素A增强CXCR4启动子活性的作用呈剂量依赖性。0~20μmol/L月腺大戟素A对MSCs无明显细胞毒性,能促进基质细胞衍生因子-1的MSCs迁移,上调MSCs的CXCR4表达水平。采用siRNA下调CXCR4表达可明显减弱月腺大戟素A促进MSCs的迁移。在体实验显示,月腺大戟素A预处理能增加心肌梗死区域内GFP-MSCs的细胞数量,增加MSCs中Akt活性,MMP2表达明显升高。结论月腺大戟素A通过上调CXCR4/Akt/MMP2信号通路促进MSCs向心肌梗死区迁移。Objective To evaluate the effects of ebracteolatain A(EA)on regulating CXCR4 expression and its mechanism of regulating the migration of mesenchymal stem cells(MSCs)to infarcted region of heart.Methods The double-luciferase reporter gene detection technology was used to evaluate the dose-effect relationship of EA in enhancing the activity of CXCR4 promoter.CCK-8 kit was used to detect the effect of EA on the cell viability of MSCs.Transwell assay was used to detect the effects of EA on the capability of MSCs migration.Western blot was used to evaluate the role of EA in regulating the CXCR4 expression of MSCs.Small interfering RNA(siRNA)technology was used to detect the role of CXCR4 in EA modulating MSCs migration.Models of myocardial infarction were established in rats and the effects of EA on the migration of green fluorescent protein(GFP)-labeled MSCs was evaluated in vivo.Western blot was used to detect EA intervention for protein kinase B(Akt)activity and matrix metalloproteinase 2(MMP2)expression in MSCs.Results EA enhanced the CXCR4 promoter activity in a dose-dependent manner.EA(0‒20μmol/L)had no obvious cytotoxicity to MSCs,could promote the migration of MSCs to SDF-1,and up-regulate the CXCR4 expression of MSCs.Using siRNA to down-regulate CXCR4 expression could significantly diminish the promoting effects of EA on MSCs migration.In vivo,pretreatment of MSCs with EA could increase cell number of GFP-MSCs in infarcted region of heart,enhance Akt activity in MSCs,and significantly increase MMP2 expression.Conclusion EA promotes MSCs to migrate to the infarcted region of heart by up-regulating the CXCR4/Akt/MMP2 signaling pathway.

关 键 词：月腺大戟素A 心肌梗死 间充质干细胞 迁移 CXCR4表达 

分 类 号：R285.5[医药卫生—中药学]