IL-6通过促进人胎盘间充质干细胞上CD73的表达调节其迁移、黏附和增殖能力  

IL-6 regulates the migration,adhesion and proliferation of human placenta-derived mesenchymal stem cells by promoting CD73 expression

作　　者：张恒超 赵楠楠 张加深 韩凯悦 赵雅萱 戚荣 栾希英[1] Zhang Hengchao;Zhao Nannan;Zhang Jiashen;Han Kaiyue;Zhao Yaxuan;Qi Rong;Luan Xiying(Department of Immunology,Binzhou Medical University,Yantai 264003,China)

机构地区：[1]滨州医学院免疫学教研室,烟台264003

出　　处：《中华微生物学和免疫学杂志》2022年第12期940-948,共9页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金(32070781);山东省自然科学基金(ZR2020MH166)。

摘　　要：目的探讨IL-6影响人胎盘间充质干细胞(human placenta-derived mesenchymal stem cells,hPMSCs)表面CD73的表达,以及调节其迁移、黏附和增殖的作用机制。方法流式细胞术(flow cytometry,FCM)和Western blot分析外源性IL-6和hPMSCs分泌的IL-6对hPMSCs上CD73表达的影响。单克隆抗体阻断和Western blot方法检测IL-6对hPMSCs中信号转导及转录激活蛋白3(signal transducer and activator of transcription 3,STAT3)磷酸化的影响。实时无标记动态细胞分析技术(real-time cellular analysis,RTCA)分析慢病毒转染后低表达CD73的hPMSCs和APCP(5′-α,β亚甲基-二磷酸腺苷)预处理后hPMSCs的迁移、黏附和增殖变化。结果FCM与Western blot检测结果显示,外源性和hPMSCs分泌的IL-6均能促进CD73在hPMSCs上的表达(P<0.001,P<0.01)。IL-6R抑制剂处理后,hPMSCs上CD73表达明显下降(P<0.01);IL-6可上调hPMSCs内STAT3磷酸化和CD73水平(P<0.05,P<0.01),而加入STAT3抑制剂后,CD73表达下降(P<0.01)。RTCA分析结果显示,敲低hPMSCs上CD73的表达后,hPMSCs的黏附和增殖能力均明显降低(P<0.01,P<0.05),迁移能力明显上升(P<0.05)。使用APCP抑制hPMSCs上CD73水解酶活性后,hPMSCs同样表现出黏附和增殖能力明显降低,而迁移能力增强(P<0.05)。结论IL-6能够通过JAK/STAT3通路增强hPMSCs上CD73的表达,进而影响其迁移、黏附和增殖能力。Objective To investigate the mechanism of IL-6 affecting the expression of CD73 on human placenta-derived mesenchymal stem cells(hPMSCs)and regulating their migration,adhesion and proliferation.Methods Flow cytometry(FCM)and Western blot were used to analyze the effects of exogenous IL-6 or IL-6 secreted by hPMSCs on the expression of CD73 on hPMSCs.The influence of IL-6 on the phosphorylation of signal transducer and activator of transcription 3(p-STAT3)in hPMSCs was detected by monoclonal antibody blocking test and Western blot.Real-time cellular analysis(RTCA)was used to analyze the changes in the migration,adhesion and proliferation of hPMSCs after knockdown of CD73 expression or APCP pretreatment.Results FCM and Western blot showed that both exogenous and autocrine IL-6 from hPMSCs promoted the expression of CD73 on hPMSCs(P<0.001,P<0.01).Moreover,CD73 expression decreased significantly with the presence of IL-6R inhibitor(P<0.01).IL-6 could up-regulate the levels of both p-STAT3 and CD73 in hPMSCs(P<0.05,P<0.01),while CD73 expression decreased after adding STAT3 inhibitor(P<0.01).RTCA showed that knockdown of CD73 expression on hPMSCs significantly inhibited the adhesion and proliferation ability of hPMSCs(P<0.01,P<0.05),but promoted the migration ability of hPMSCs(P<0.05).Similarly,inhibiting the hydrolase activity of CD73 on hPMSCs by APCP also resulted in a significant decrease in the adhesion and proliferation capacities of hPMSCs,and an increase in the migration capacity of hPMSCs(P<0.05).Conclusions IL-6 enhanced the expression of CD73 on hPMSCs via the JAK/STAT3 pathway,thus affecting the migration,adhesion and proliferation of hPMSCs.

关 键 词：间充质干细胞 胎盘 IL-6 CD73 迁移 黏附 增殖 

分 类 号：R392[医药卫生—免疫学]