HucMSC-ex通过Let-7b调控TGF-βR_1抑制肝星状细胞胶原合成  

HucMSC-ex inhibit collagen synthesis of hepatic stellate cells through Let-7b mediated TGF-βR_1 downregulation

作　　者：蔡梦洁 钱嘉莉 陈逸菲 王岩金 赵婷[1] 刘锦文 沈松[2] 严永敏 CAI Meng-jie;QIAN Jia-li;CHEN Yi-fei;WANG Yan-jin;ZHAO Ting;LIU Jin-wen;SHEN Song;YAN Yong-min(School of Medicine;School of Pharmacy,Jiangsu University,Zhenjiang Jiangsu H2013,China)

机构地区：[1]江苏大学医学院,江苏镇江212013 [2]江苏大学药学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2018年第6期478-482,共5页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(81670549);江苏省高校自然科学基金资助项目(15KJB350001)

摘　　要：目的:探讨人脐带间质干细胞来源的外泌体(human umbilical cord mesenchymal stem cells derived exosome,Huc MSC-ex)对肝星状细胞LX-2胶原合成的影响及作用机制。方法:提取Huc MSC-ex,采用透射电镜分析Huc MSC-ex形态,蛋白质印迹法检测特异性标志蛋白CD9和CD63的表达;建立TGF-βR_1诱导的人肝星状细胞(hepatic stellate cells,HSC) LX-2细胞株活化模型,分别加入25,50和100μg/m L Huc MSC-ex共培养,另设对照组(常规培养),qRT-PCR检测4组Col1、Col3、TGF-βR_1和Let-7b mRNA表达,免疫印迹法检测对应的蛋白表达; Let-7b过表达转染LX-2细胞株,qRT-PCR和免疫印迹法检测LX-2中TGF-βR_1、αSMA蛋白和mRNA表达。结果:Huc MSC-ex是30～100 nm的膜性囊泡,表达外泌体的特异标志CD9和CD63。与对照组相比,Huc MSC-ex处理组中Col1、Col3和TGF-βR_1表达明显下调(P均<0. 001),而Let-7b表达明显上调(P均<0. 001); Let-7b过表达显著抑制LX-2细胞中TGF-βR_1和α-SMA的表达(P均<0. 05)。结论:Huc MSC-ex转运Let-7b到肝星状细胞,调控肝星状细胞TGF-βR_1表达,抑制肝星状LX-2细胞胶原合成。Objective: To study the effect of human umbilical cord mesenchymal stem cells derived exosomes( HucMSC-ex) on collagen synthesis in hepatic stella cell line( LX-2) and its mechanism.Methods: HucMSC-ex was identified by transmission electron microscopy for morphology and western blotting for exosomal markers CD9 and CD63 expression. The mRNA levels of Col1,Col3,TGF-βR1 and Let-7 b in LX-2 cells were detected by qRT-PCR. The level of TGF-βR1 protein in LX-2 cells was detected by western blotting. Results: HucMSC-ex are membrane vesicles with a diameter of 30-100 nm and express exosomal markers CD9 and CD63. Compared with the control group,HucMSC-ex significantly reduced Col1,Col3 and TGF-βR1 expression( all P < 0. 001),promoted Let-7 b expression( all P <0.001). Let-7b mimics transfection down-regulated TGF-βR1 and α-SMA expression in LX-2 cells( both P< 0.05).Conclusion: HucMSC-ex may inhibit LX-2 activation through delivering Let-7 b to regulate TGF-βR1 expression in hepatic stellate cells.

关 键 词：Let-7b 人脐带间质干细胞来源的外泌体 肝星状细胞 

分 类 号：R393[医药卫生—基础医学]