miR-125b-5p修饰脐带间充质干细胞介导JAK2/STAT3信号通路对系统性红斑狼疮的免疫调节作用  

Immunomodulatory effect of miR-125b-5p modified umbilical cord mesenchymal stem cells mediated JAK2/STAT3 signaling pathway on systemic lupus erythematosus

作　　者：武志慧 丁丽丽 胡明智 王永福[2] 王慧[2] 张伟[1] 孙晓林 Wu Zhihui;Ding Lili;Hu Mingzhi;Wang Yongfu;Wang Hui;Zhang Wei;Sun Xiaolin(Central Laboratory,Key Laboratory of Autoimmunology,The First Affiliated Hospital of Baotou Medical College,Baotou 014010;Dept of Rheumatology,The First Affiliated Hospital of Baotou Medical College,Baotou 014010)

机构地区：[1]包头医学院第一附属医院中心实验室(内蒙古自治区自体免疫学重点实验室),包头014010 [2]包头医学院第一附属医院风湿免疫科,包头014010

出　　处：《安徽医科大学学报》2023年第1期28-36,共9页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81860294,81860295);内蒙古自治区自然科学基金(编号:2019MS08055,2021MS08045);内蒙古自治区科技计划项目(编号:201802089,2019GG052)。

摘　　要：目的研究miR-125b-5p修饰脐带间充质干细胞(UC-MSCs)介导JAK2/STAT3信号通路对系统性红斑狼疮(SLE)的免疫调节作用。方法UC-MSCs的分离、培养及鉴定;密度梯度离心法分离SLE患者外周血单个核细胞(PBMCs),实时荧光定量PCR检测与各组UC-MSCs共培养48 h后的PBMCs中干扰素γ(IFN-γ)、白细胞介素4(IL-4)、白细胞介素17A(IL-17 A)、叉头状转录因子3(Foxp 3)基因的相对表达量;Western blot检测治疗后MRL/lpr小鼠肾组织中Janus激酶2(JAK2)、磷酸化Janus激酶2(p-JAK2)、信号转导与转录激活因子3(STAT3)、磷酸化信号转导与转录激活因子3(p-STAT3)、白细胞介素18(IL-18)蛋白的相对表达量。结果与PBMCs组相比,UC-MSCs+miR-125b-5p组、UC-MSCs+miR-NC组、UC-MSCs组的IFN-γ、IL-17 A的表达和辅助性T细胞17(Th 17)/调节性T细胞(Treg)的细胞比例显著下调(P<0.01),UC-MSCs+miR-125b-5p组、UC-MSCs+miR-NC组的辅助性T细胞1(Th 1)/辅助性T细胞2(Th 2)细胞比例下调(P<0.05);与MRL/lpr小鼠未处理组相比,UC-MSCs+miR-125b-5p组治疗后的MRL/lpr小鼠肾组织中p-JAK2/JAK2、p-STAT3(s 705)/STAT3、IL-18的表达水平显著下调(P<0.01);UC-MSCs+miR-NC组、UC-MSCs组治疗后的MRL/lpr小鼠肾组织中IL-18的表达水平显著下调(P<0.01)。结论miR-125b-5p基因修饰UC-MSCs后,对调节SLE患者PBMCs中Th2细胞亚群分化以及MRL/lpr小鼠肾组织中p-JAK 2/JAK 2、p-STAT 3(s 705)/STAT 3、IL-18的表达起到了协同作用。miR-125b-5p基因修饰的UC-MSCs可能通过介导JAK2/STAT3通路对SLE起到免疫调节作用。Objective To investigate the immunomodulatory effects of umbilical cord mesenchymal stem cells(UC-MSCs)modified by miR-125b-5p through JAK2/STAT3 pathway on systemic lupus erythematosus(SLE).Methods UC-MSCs were isolated and cultured under aseptic conditions;Peripheral blood mononuclear cells(PBMCs)were separated from SLE patients by density gradient centrifugation.The relative expressions of IL-17A,Foxp3,IFN-γ,IL-4 genes in PBMCs cultured with UC-MSCs for 48 h were detected by RT-qPCR;The relative expressions of JAK2,p-JAK2,STAT3,p-STAT3 and IL-18 proteins in kidney tissues of MRL/lpr mice were detected by western blot.Results Compared with the PBMCs culture group,the expression of IFN-γand IL-17A and the proportion of Th17/Treg cells were significantly down-regulated in UC-MSCs+miR-125b-5p group,UC-MSCs+miR-NC group and UC-MSCs group(P<0.01),the expression of the proportion of Th1/Th2 cells(P<0.05)was down-regulated in UC-MSCs+miR-125b-5p group and UC-MSCs+miR-NC group;Compared with the untreated group of MRL/lpr mice,the relative expressions of JAK2,p-JAK2,STAT3,p-STAT3 and IL-18 proteins in kidney tissues of MRL/lpr mice were significantly down-regulated in UC-MSCs+miR-125b-5p group(P<0.01),and the relative expressions of IL-18 proteins was significantly down-regulated in UC-MSCs+miR-NC group and UC-MSCs group(P<0.01).Conclusion miR-125b-5p plays a synergistic role in UC-MSCs,which regulates the differentiation of Th2 cells in PBMCs of SLE patients and the relative expressions of p-JAK2/JAK2,p-STAT3/STAT3,and IL-18 proteins in kidney tissues of MRL/lpr mice.UC-MSCs modified by miR-125b-5p may play immunomodulatory effect on SLE by JAK2/STAT3 signaling pathway.

关 键 词：基因修饰 脐带间充质干细胞 系统性红斑狼疮 JAK2/STAT3信号通路 

分 类 号：R392[医药卫生—免疫学] R443.8[医药卫生—基础医学] R593.2