CAJ | 학술논문

目的研究小鼠心脏移植排斥反应中小分子RNA494(miR-494)与辅助性T淋巴细胞17(TH 17细胞)分化的相关性.方法分别建立颈部异位心脏移植模型:对照组,供者和受者均为C57BL/6小鼠;实验组,供者为Balb/c小鼠,受者为C57BL/6小鼠.利用逆转录聚合酶链反应(RT-PCR)检测移植物中miR-494与IL-17A mRNA的表达.分离移植物中CD4+T淋巴细胞、CD8+T淋巴细胞及CD45+细胞,检测各组细胞中miR-494与IL-17A mRNA的表达.体外分离C57BL/6小鼠脾细胞,利用免疫磁珠法进行CD4+T淋巴细胞分选,定向分化为TH 1、TH2、TH 17和调节性T淋巴细胞(Treg细胞)亚群,采用RT-PCR检测各细胞亚群IL-17AmRNA及miR-494的表达.结果小鼠心脏移植术后7d,实验组移植物中IL-17A mRNA表达高于对照组,而miR-494的表达低于对照组(均P＜0.05).移植物CD4+T淋巴细胞IL-17AmRNA表达高于对照组,而miR-494的表达降低(均P＜0.05).体外培养TH 17细胞亚群中,miR-494的表达显著低于TH 1、TH2和Treg细胞.结论 miR-494可能与移植排斥反应中TH 17细胞分化密切相关,miR-494可能通过调控TH17细胞在移植排斥反应中发挥作用.
목적 연구소서심장이식배척반응중소분자RNA494(miR-494)여보조성T림파세포17(TH 17세포)분화적상관성.방법 분별건립경부이위심장이식모형:대조조,공자화수자균위C57BL/6소서;실험조,공자위Balb/c소서,수자위C57BL/6소서.이용역전록취합매련반응(RT-PCR)검측이식물중miR-494여IL-17A mRNA적표체.분리이식물중CD4+T림파세포、CD8+T림파세포급CD45+세포,검측각조세포중miR-494여IL-17A mRNA적표체.체외분리C57BL/6소서비세포,이용면역자주법진행CD4+T림파세포분선,정향분화위TH 1、TH2、TH 17화조절성T림파세포(Treg세포)아군,채용RT-PCR검측각세포아군IL-17AmRNA급miR-494적표체.결과 소서심장이식술후7d,실험조이식물중IL-17A mRNA표체고우대조조,이miR-494적표체저우대조조(균P＜0.05).이식물CD4+T림파세포IL-17AmRNA표체고우대조조,이miR-494적표체강저(균P＜0.05).체외배양TH 17세포아군중,miR-494적표체현저저우TH 1、TH2화Treg세포.결론 miR-494가능여이식배척반응중TH 17세포분화밀절상관,miR-494가능통과조공TH17세포재이식배척반응중발휘작용.
Objective To investigate correlation between microRNA (miR-494) and TH 17 cell differentiation in murine cervical heterotopic cardiac transplant model.Method The heterotopic cardiac transplant models of Balb/c→C57BL/6 mice were established as experimental group,and those of C57BL/6→C57BL/6 mice as control group.Real time-polymerase chain reaction(PCR) was used to detect miR-494 and interleukin(IL)-17A mRNA expression in the grafts.CD4+ T cells,CD8+ T cells and CD45+ myeloid cells were isolated from the grafts,and miR-494 and IL-17 mRNA expression was detected.In vitro,lymphocytes in the spleen from C57BL/6 mice were harvested,and CD4+ T cells were isolated with MACS and then stimulated to TH 1,TH 2,TH 17,Treg subset cells.The expression of IL-17A mRNA and miR-494 in different T subsets was examined by Reverse transcription-polymerase chain reaction(RT-PCR).Result Two grafts from each study group were harvested on the 7th day post-transplantation.In experimental group,the IL-17A mRNA expression was increased,while the expression of miR-494 was decreased as compared with control group with the difference being significant between two groups.The expression of IL-17A rnRNA in CD4+ T cells of the grafts was significantly increased,while that expression of miR-494 was decreased.In vitro,the expression of miR-494 in TH 17 cells was significantly lower than that in TH 1,TH 2 and Treg cells.Conclusion miR-494 is related closely to TH 17 cells differentiation in the transplant rejection,which may play a role in transplant rejection through regulating TH 17 cells.