miR-146a下调小鼠TRAF-6并减少胸腺基质细胞凋亡率的实验研究  

Overexpression of microRNA miR-146a diminish the apoptosis ratio of the thymic stromal cell by inhib-iting the expression of the target gene TRAF-6

作　　者：刘学[1] 王亚君[1] 杜红梅[1] 朱喜科[1] Liu Xue;Wang Yajun;Du Hongmei;Zhu Xike(Medical Research Center, Shengjing Hospital of China Medical University, Shenyang 110004, China)

机构地区：[1]中国医科大学附属盛京医院实验研究中心,沈阳110004

出　　处：《国际免疫学杂志》2019年第2期121-126,共6页International Journal of Immunology

基　　金：国家自然科学基金(81270430).

摘　　要：目的探讨miR-146a在胸腺发育及增龄性萎缩中的作用.方法转染miR-146a模拟物到小鼠胸腺基质细胞(thymic stromal cell,TSC)中,48 h后使用逆转录-聚合酶链反应(reverse transcrip-tion-polymerase chain reaction,RT-PCR)技术测其含量,观察有无成功过表达;72 h后使用Western blot检测其靶基因肿瘤坏死因子受体相关因子6(tumor necrosis factor receptor-associated factor 6,TRAF-6)含量,并采用流式细胞仪观察TSC凋亡率的变化.结果在转染48 h后TSC中miR-146a含量明显升高(mimics组为192.95±54.64,对照组为1,P<0.05);72 h后其靶基因TRAF-6在mimics组中含量相比对照组明显降低(mimics组为0.50±0.25,对照组为1,P<0.05);转染组胸腺基质细胞72 h凋亡率较对照组降低(mimics组为13.90%±1.84%,对照组为22.83%±3.43%,P<0.05).结论 miR-146a随年龄在胸腺基质中含量下降与胸腺萎缩是相关性,考虑其对靶基因的抑制减弱所引起,并且与靶基因TRAF-6能促进细胞凋亡有一定关系.Objective To explore the influence of microRNA miR-146a in the development and aging atrophy of thymus. Methods miR-146a was overexpressed in the thymic stromal cell(TSC)by transfection with miR-146a mimic. After 48 hours of the transfection,the miR-146a content was detected by reverse tran-scription polymerase chain reaction(RT-PCR). After 72 hours of the transfection,the protein level of target gene tumor necrosis factor receptor-associated factor 6(TRAF-6)was tested by Western Blot method and the apoptosis ratio of TSC was assayed by flow cytometry. Results The expression of miR-146a was significantly higher than those in the negative control group[(192. 95 ± 54. 64)vs. 1]P < 0. 05)after 48 hours of the trans-fection. The TRAF-6 protein level was decreased[(0. 50 ± 0. 25)vs. 1]and the apoptosis ratio[(13. 90%± 1. 84%)vs.(22. 83%± 3. 44%)]was diminished in the mimics group by comparing with the negative control group(both P < 0. 05). Conclusion Overexpression of miR-146a could diminish the apoptosis ratio of TSC by inhibiting the expression of the target gene TRAF-6.

关 键 词：胸腺 胸腺基质细胞 MIR-146A 肿瘤坏死因子受体相关因子6 凋亡 

分 类 号：R392.12[医药卫生—免疫学]