原因不明复发性自然流产患者外周血IL-6、let-7d-5p水平变化及其关系

摘要

Objective To investigate the changes of IL-6 and microRNA ( miRNA) let-7d-5p in the peripheral blood of patients with unexplained recurrent spontaneous abortion ( URSA) and to explore the relationships between them. Methods Thirty cases of normal pregnant women ( normal pregnancy group) and 30 cases of URSA patients ( URSA group) were included in this study. The serum and peripheral blood mononuclear cells ( PBMCs) of all subjects were taken from each group. The protein level of IL-6 in plasma was detected by ELISA, and let-7d-5p expression was detected by q PCR.Target Scan 7. 1 software was used to predict the binding site between let-7d-5p and IL-6; besides, 3' untranslated region ( 3' UTR) luciferase reporter assay was used to confirm the binding between them. The 293 T cells were divided into the intervention group and control group, and cells were transfected with let-7d-5p mimics ( blank negative control) and widetype or mutant IL-6 mRNA 3' UTR fluorescent reporter gene pmir GLO vector, respectively. Luciferase reporter gene activity was detected by using dual luciferase reporter gene system. In addition, 293 T cells were divided into 3 groups. The cells in the overexpression group and the inhibition group were transfected with leto-7d-5p mimics and inhibitor by using Lipofectamine 2000, respectively, and the cells in the blank control group were transfected with negative control primers. The cells were harvested 24 h after transfection. Then, the expression of IL-6 mRNA was detected by q PCR, and the protein level of IL-6 was detected by ELISA. Results Compared with the normal pregnancy group, the serum IL-6 protein level increased significantly while the let-7d-5 expression in the PBMCs decreased significantly in the URSA group ( both P 0. 05) . Compared with the blank control group, IL-6 mRNA expression and IL-6 protein level decreased in the overexpression group, but increased in the inhibition group ( both P 0. 05) .与空白对照组比较, 过表达组细胞中IL-6 mRNA表达及细胞上清液中IL-6水平降低 (P均<0. 05) , 抑表达组细胞中IL-6 mRNA表达及细胞上清液中IL-6水平升高 (P均<0. 05) .结论 URSA患者血清IL-6蛋白水平升高, 而PBMC中let-7d-5p表达降低; let-7d-5p与IL-6 mRNA 3'UTR结合并抑制IL-6表达, 靶向let-7d-5p调控IL-6表达可能成为URSA治疗的有效途径.