LncRNA XIST沉默对非酒精性脂肪肝小鼠T细胞免疫功能的作用机制  

The effect of LncRNA XIST silencing on T cell immune function and its mechanism in nonalcoholic fatty liver mice

作　　者：方金鸣 FANG Jinming(Quality Management Office of Wuhan Fourth Hospital,Wuhan 430033,China)

机构地区：[1]武汉市第四医院质量管理办公室,430033

出　　处：《免疫学杂志》2023年第9期769-776,共8页Immunological Journal

基　　金：武汉市卫生健康委科研基金(WX19Y10)。

摘　　要：目的探讨长链非编码RNA XIST(LncRNA XIST)调节微小RNA-137-3p(miR-137-3p)/Krüppel样转录因子2(KLF2)轴对非酒精性脂肪肝(NAFLD)小鼠T细胞免疫功能的影响。方法建立NAFLD小鼠模型,检测脾脏CD4+T、CD8+T细胞及细胞中LncRNA XIST表达,将CD4+T细胞分为si-NC组、si-XIST组、si-XIST+anti-NC组及si-XIST+anti-miR-137-3p组,分别检测LncRNA XIST、miR-137-3p和KLF2表达水平、细胞增殖与凋亡;将各组转染的CD4+T细胞与人肝细胞HL-7702共培养,分别检测TGF-β1、IL-10、IFN-γ、IL-17水平及Th17、Treg细胞比例;双荧光素酶报告基因实验验证LncRNA XIST、KLF2与miR-137-3p靶向关系。结果NAFLD组小鼠CD4+T细胞比例显著升高,CD8+T细胞比例显著降低(P<0.05),且LncRNA XIST在CD4+T细胞中表达水平显著升高(P<0.05);抑制LncRNA XIST表达可显著抑制CD4+T细胞增殖,诱导细胞凋亡,诱导CD4+T细胞TGF-β1、IL-10分泌,抑制IFN-γ、IL-17水平,升高Treg细胞比例,降低Th17细胞比例;抑制miR-137-3p表达可逆转抑制LncRNA XIST表达对CD4+T细胞的影响;双荧光素酶报告基因实验证实,LncRNA XIST可调控miR-137-3p表达,KLF2是miR-137-3p下游靶点。结论LncRNA XIST在NAFLD小鼠CD4+T细胞中表达水平升高,抑制LncRNA XIST表达可通过调节miR-137-3p/KLF2信号轴,调节炎性因子分泌,升高Treg细胞比例,降低Th17细胞比例。This study was designed to investigate the influence of long non-coding RNA XIST(LncRNA XIST)on T cell immune function in mice with nonalcoholic fatty liver disease(NAFLD)by regulating microRNA-137-3p(miR-137-3p)/Krüppel like transcription factor 2(KLF2)axis.NAFLD mouse model was established,and the levels of CD4+T and CD8+T cells,as well as LncRNA XIST expression in these cells were detected.CD4+T cells were grouped into si-NC group,si-XIST group,si-XIST+anti NC group,and si-XIST+anti miR-137-3p group,then the expression levels of LncRNA XIST,miR-137-3p and KLF2,the cell proliferation and apoptosis were detected respectively.The transfected CD4+T cells in each group were co cultured with human hepatocyte HL-7702,and the levels of TGF-β1,IL-10,IFN-γ,IL-17 and the proportion of Th17 and Treg cells were detected respectively;double luciferase reporter gene experiment was applied to verify the targeting relationship of miR-137-3p with LncRNA XIST or KLF2.Data showed that in NAFLD group,the percentage of CD4+T cells increased obviously,while the percentage of CD8+T cells decreased obviously(P<0.05),the expression level of LncRNA XIST in CD4+T cells increased obviously(P<0.05).Inhibiting the expression of LncRNA XIST could obviously inhibit the proliferation of CD4+T cells,induce apoptosis,induce the secretion of TGF-β1 and IL-10 in CD4+T cells,inhibit the levels of IFN-γand IL-17,increase the proportion of Treg cells,and reduce the proportion of Th17 cells.Inhibition of miR-137-3p expression could reverse the effect of LncRNA XIST inhibition on CD4+T cells;the double luciferase reporter gene experiment confirmed that LncRNA XIST could regulate the expression of miR-137-3p,and KLF2 was the downstream target of miR-137-3p.All data suggested that the expression level of LncRNA XIST in CD4+T cells of NAFLD mice is increased;and inhibition of LncRNA XIST expression could adjust the miR-137-3p/KLF2 signal axis to regulate the secretion of inflammatory factors,increase the proportion of Treg cells and reduce the propor

关 键 词：长链非编码RNA XIST 微小RNA-137-3p Krüppel样转录因子2 非酒精性脂肪肝 细胞免疫 

分 类 号：R575.5[医药卫生—消化系统]