miR-7敲减CD4+T细胞过继转输对小鼠急性肝损伤模型的影响  

Effect of adoptive transfer of CD4^+ T cells with miR-7 knockdown on acute liver injury in mice

作　　者：岳东旭[1] 赵娟娟[1] 陈慧子 丁涛[1] 胡琳 潘飞飞 郭萌萌[1] 陈超[1] 徐林[1] YUE Dong-xu;ZHAO Juan-juan;CHEN Hui-zi;DING Tao;HU Lin;PAN Fei-fei;GUO Meng-meng;CHEN Chao;XU Lin(Department of Immunology,Zunyi Medical University,Talent Base of Biological Therapy of Guizhou Province,Special Key Lab of Gene Detection and Treatment of Guizhou Province,Zunyi 563099,China)

机构地区：[1]遵义医学院免疫学教研室暨贵州省生物治疗人才基地,贵州省基因检测与治疗特色重点实验室,贵州遵义563099

出　　处：《中国病理生理杂志》2018年第9期1660-1665,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.31760258);贵州省高层次创新人才计划[黔科合人才(2016)4031号];遵义医学院优秀青年人才计划项目(No.15ZY-001)。

摘　　要：目的:观察过继转输微小RNA-7(microRNA-7,miR-7)敲减(knockdown,KD)CD4^+ T细胞对小鼠急性肝损伤模型的影响,并探讨其意义。方法:利用磁珠分选技术分别获得正常野生型(WT)小鼠和miR-7KD小鼠脾脏中CD4^+ CD62L^+ T细胞;CFSE染色标记后,按每只小鼠2×10~6个细胞,经尾部静脉注射给同系WT小鼠,并利用伴刀豆球蛋白A建立急性肝损伤模型;观察过继转输后2组小鼠肝脏的形态、重量及其重量指数的变化;HE染色观察小鼠肝脏组织的病理学变化;real-time PCR检测小鼠肝脏组织中凋亡相关分子Bax和P53的表达;流式细胞术检测小鼠肝脏组织中CFSE标记的CD4^+ T细胞活化相关膜分子CD62L表达水平以及细胞因子白细胞介素4(IL-4)和干扰素γ(IFN-γ)的表达变化。结果:与对照组相比,过继转输miR-7KD小鼠CD4^+ CD62L^+ T细胞组(miR-7KD转输组)小鼠的肝脏重量明显减轻(P<0.01),但重量指数显著增加(P<0.05);HE染色显示miR-7KD转输组的肝脏细胞损伤增加;real-time PCR结果显示miR-7KD转输组肝脏组织中凋亡相关细胞分子Bax和P53的相对表达水平均明显升高(P<0.05);流式细胞术检测结果进一步显示肝脏组织中的CFSE^+细胞活化相关分子CD62L的表达水平明显降低(P<0.01),细胞因子IFN-γ的表达水平明显增加(P<0.05),而IL-4的表达水平显著降低(P<0.01)。结论:过继转输miR-7敲减CD4^+ T细胞可显著加重急性肝损伤模型小鼠的肝组织损伤。这为后续深入探讨急性肝损伤的发生机制提供了重要的实验基础。AIM:To study the effect of adoptive transfer of CD4+T cells with microRNA-7(miR-7)knockdown(KD)on mouse acute liver injury model and to investigate its significance.METHODS:CD4+CD62L+T cells were purified from the spleen of normal wild-type(WT)mice and miR-7KD mice by magnetic bead sorting,and were stained with CFSE.These 2×10 6 CFSE-labeling cells were injected into normal mice via tail vein,and then the mouse acute liver injury model was induced by intraperitoneal injection of 30 mg/kg concanavalin A.After 72 h,the appearance,weight and weight index of the liver were investigated.The pathological change of the liver tissues was observed by HE staining.Real-time PCR was used to examine the mRNA expression of Bax and P53.The expression levels of CD62L,interleukin-4(IL-4)and interferon-γ(IFN-γ)in the CD4+T cells were analyzed by flow cytometry.RESULTS:We found that the liver tissue became lighter,and the weight(P<0.01)and weight index(P<0.05)were changed significantly in miR-7KD mice compared with control group.Moreover,HE staining showed that the liver cell damage was increased in the liver of miR-7KD mice.Meanwhile,the expression levels of Bax and P53 were significantly increased in miR-7KD group(P<0.05).The percentage of CD62L in CD4+T cells was significantly decreased(P<0.01)in miR-7KD mice,with high expression of IFN-γ(P<0.05)and low expression of IL-4(P<0.01)in CD4+T cells.CONCLUSION:These findings suggest that miR-7 knockdown significantly promotes the pathology of CD4+T cell-mediated acute liver injury,which provides a preliminary experimental basis for further exploration on the mechanism of acute liver injury occurrence.

关 键 词：微小RNA-7 急性肝损伤 CD4+T细胞 细胞因子 细胞凋亡 

分 类 号：R394-33[医药卫生—医学遗传学] R363[医药卫生—基础医学]