[Pathogenesis of Immune Thrombocytopenic Purpura (ITP) by MiRNA-30a-Mediated Th17 Cell Differentiation].

• DOI: 10.19746/j.cnki.issn.1009-2137.2020.02.039
• Publication Date: 2020-04-01
• Authors: Xiaofang Wu; Jianqin Li; Shaoyan Hu; Zhaoyue Wang; Lan Dai; Chenmei Mao; Jing Ling
• Keywords: Pathogenesis; Thrombocytopenic purpura; Splenocyte

To investigate whether miRNA-30a is involved in the pathogenesis of ITP by affecting differentiation Th17 cells, and to explore its possible mechanism through verification target gene SOCS3 for prediction miRNA-30a.Firstly, a chronic mouse model was established. The expression RORγt spleen mononuclear cells were detected their correlation analyzed. Secondly, luciferase vector containing 3'UTR green fluorescent miRNA constructed. Luciferase fluorescence detection, real-time quantitative PCR (qPCR) Western blot used verify miRNA-30a.The platelet count mice experimental group decreased below 20% normal ones after 48 hours injection anti-mouse serum (APS), which maintained 14 days at least; higher than those control group(P＜0.05), moreover, there positive between them (r=0.54); activity PMDH-GFP-miRNA-30a pMIR-report-UTR significantly lower that PMDH-GFP empty plasmid pMIR-report-UTR(P＜0.05); mRNA protein level not different from group.Chronic has been established successfully; increase, positively correlated with RORγt, contribute cells; able bind site miRNA-30a, but might be functional gene.miRNA-30a通过介导Th17细胞分化影响免疫性血小板减少性紫癜发病的初步探讨.探讨miRNA-30a是否通过影响Th17细胞分化参与ITP发病；通过对miRNA-30a预测靶基因SOCS3的验证，进一步探讨miRNA-30a参与ITP发病的可能机制.建立慢性ITP小鼠模型，检测其脾脏单个核细胞中miRNA-30a、 RoRγt的表达并分析两者相关性；构建含靶基因mRNA 3′UTR的荧光素酶表达载体及含有miRNA的绿色荧光载体，通过Luciferase荧光检测、实时荧光定量PCR（qPCR）、Western blot验证SOCS3是否为miRNA-30a的靶基因.实验组小鼠血小板计数在注射抗小鼠血小板血清（APS）48 h后下降至正常的20%，且至少可维持14 d。实验组小鼠脾脏单个核细胞中miRNA-30a及RoRγt表达较对照组升高（P＜0.05），且两者存在正性相关（r=0.54）。pMDH-GFP-miRNA-30a与pMIR-report-UTR实验组的荧光素酶的活性明显低于pMDH-GFP空质粒与pMIR-report-UTR的对照组（P＜0.05）。转染miRNA-30a质粒的EL4细胞中SOCS3在mRNA及蛋白水平与对照组均无差异.miRNA-30a在ITP小鼠脾脏单个核细胞中表达升高，且与RORγt表达呈正相关性，它可能通过影响Th17细胞分化参与ITP发病；SOCS3能与miRNA-30a靶位点结合，却不是其功能靶基因.