自身免疫糖尿病患者外周血单个核细胞miR-142和miR-143表达的研究

Expression of miR-142 and miR-143 in peripheral blood mononuclear cells from patients with autoimmune diabetes

目的探讨自身免疫糖尿病患者(ADM),包括经典1型糖尿病(T1DM)和成人隐匿性自身免疫糖尿病(LADA),以及2型糖尿病患者(T2DM)和健康对照外周血单个核细胞(PBMC)的microRNA(miRNA)表达水平。方法收集2015年1月至2016年12月在中南大学湘雅二医院就诊的糖尿病患者及健康体检中心体检的健康对照人群,研究对象分为两批,分别进行高通量测序初筛差异表达的miRNA,以及对miR-142和miR-143的表达进行实时定量聚合酶链反应(RT-qPCR)验证。结果(1)T1DM患者、LADA患者、T2DM患者以及健康对照的PBMC中miRNA的表达存在差异;(2)与T2DM患者和健康对照相比,LADA和T1DM患者PBMC中的miR-142-5p表达下调,而miR-143-3p表达上调;(3)RT-qPCR验证实验显示,miR-142-5p在LADA患者中表达水平较T2DM患者显著下调。miR-143-3p在T1DM患者和LADA患者中表达量较T2DM患者和健康对照有升高趋势,但差异无统计学意义。结论(1)T1DM患者、LADA患者、T2DM患者以及健康对照的PBMC中miRNA的表达存在差异;(2)miR-142-5p和miR-143-3p的异常表达可能通过影响细胞凋亡和免疫细胞分化参与ADM发生发展。

Objective To investigate the microRNA ( miRNA ) expression level of peripheral blood mononuclear cell ( PBMC) in autoimmune diabetes mellitus ( ADM) which includes type 1 diabetes mellitus ( T1 DM) and latent autoimmune diabetes in adults ( LADA), T2DM patients, and matched healthy individuals. Methods Patients of T1DM, LADA, and T2DM were recruited in the Second Xiangya Hospital of Central South University from January 2015 to December 2016. The subjects were divided into two groups. The first group was used for high- throughput screening of differentially expressed microRNAs. The second group was used to validate the expression of miR-142-5p and miR-143-3p by real-time quantitative polymerase chain reaction ( RT-qPCR). Results ( 1 ) The different miRNA expression patterns of PBMC were found among T1 DM patients, LADA patients, T2DM patients, and health individuals.(2 ) Compared with T2DM patients and healthy controls, LADA and T1DM patients had down- regulated PBMC miR-142-5p expression, and up-regulated miR-143-3p expression.(3) RT-qPCR validation showed that the expression of miR-142-5p in LADA patients was significantly lower than that in T2DM patients (0.30 ±0.24 vs 1.33 ± 1.29, P<0.05). The expression of miR-143-3p in T1 DM and LADA was higher than that in T2DM and health individuals. However, no significant differences were found. Conclusion The miRNA expression patterns are different in the PBMC of T1DM patients, LADA patients, T2DM patients, and healthy individuals;the abnormal expressions of miR-142-5p and miR-143-3p may participate in the development of ADM by affecting apoptosis and immune cell differentiation.