微小RNA-17-5p在小儿肾病综合征发病中的作用及其机制研究

Role of microRNA-17-5p in the pathogenesis of pediatric nephrotic syndrome and related mechanisms

摘要:

目的 分析微小RNA-17-5p（miR-17-5p）在小儿肾病综合征（NS）发病中的意义及其通过激活素A（ActA）/Smads通路对肾足细胞凋亡的影响。方法 选取2018年3月至2019年3月收治的55例NS患儿为NS组，另选取50例同期体检的健康儿童为正常对照组，比较两组外周血miR-17-5p表达情况。培养人肾足细胞株，分别转染含miR-17-5p反义寡核苷酸重组质粒（抑制组）、含无意义随机序列的对照载体（阴性对照组），未处理的人肾足细胞为空白组。比较各组细胞凋亡情况，以及转染后细胞miR-17-5p、ActA mRNA、Smads mRNA和相关蛋白表达情况。结果 NS组外周血miR-17-5p水平高于正常对照组（P < 0.001）。与空白组和阴性对照组比较，抑制组细胞凋亡率、miR-17-5p相对表达量更低，ActA、Smad2、Smad3三者的mRNA及蛋白相对表达量更高（P < 0.001）。结论 miR-17-5p在小儿NS外周血的含量升高，低表达miR-17-5p能抑制人肾足细胞的凋亡，其机制可能与上调ActA、Smad2、Smad3三者的mRNA及蛋白表达有关。

Abstract:

Objective To study the role of microRNA-17-5p (miR-17-5p) in the pathogenesis of pediatric nephrotic syndrome (NS) and its effect on renal podocyte apoptosis via the activin A (ActA)/Smads pathway. Methods An analysis was performed on 55 children with NS (NS group) who were admitted from March 2018 to March 2019. Fifty healthy children who underwent physical examination during the same period of time were enrolled as the control group. The mRNA expression of miR-17-5p in peripheral blood was measured and compared between the two groups. Human renal podocytes were transfected with antisense oligonucleotide recombinant plasmid containing miR-17-5p (inhibition group) or control vector containing nonsense random sequence (negative control group), and untreated human renal podocytes were used as the blank group. These groups were compared in terms of cell apoptosis and the mRNA and protein expression of miR-17-5p, ActA, and Smads after transfection. Results The NS group had a significantly higher level of miR-17-5p in peripheral blood than the control group (P < 0.001). Compared with the blank and negative control groups, the inhibition group had significantly lower apoptosis rate and relative mRNA expression of miR-17-5p and significantly higher relative mRNA and protein expression of ActA, Smad2, and Smad3 (P < 0.001). Conclusions There is an increase in the content of miR-17-5p in peripheral blood in children with NS. Low expression of miR-17-5p can inhibit the apoptosis of human renal podocytes, which may be associated with the upregulation of the mRNA and protein expression of ActA, Smad2 and Smad3.