Ni Fen-Fen, Liu Guang-Lei, Jia Shi-Lei, Chen Ran-Ran, Liu Li-Bing, Li Cheng-Rong, Yang Jun, Gao Xiao-Jie
Department of Nephrology, Shenzhen Children's Hospital, Shenzhen, China.
The Fifth Affiliated (Zhuhai) Hospital of Zunyi Medical University, Zhuhai, China.
Front Pediatr. 2021 Apr 21;9:651544. doi: 10.3389/fped.2021.651544. eCollection 2021.
We investigated the pathogenesis of idiopathic nephrotic syndrome (INS) by measuring the effects two specific miRNAs on Th2 cells in children with this disease. After informed consent, we enrolled 20 children with active INS before steroid initiation, 20 children with INS in remission after steroid therapy, and 20 age-matched healthy controls. Flow cytometry was used to measure the levels of Th2 cells and a cytometric bead array was used to measure the levels of IgE, interleukin (IL)-4, and IL-13. RT-PCR was used to measure the levels of miR-24 and miR-27 in CD4TCD25 cells. PBMCs were isolated using Ficoll density gradient centrifugation, and transfected with different mimic or inhibitor miRNAs. RT-PCR was used to measure the expression of different RNAs, and flow cytometry was used to determine the percentage of Th2 cells. Relative to healthy controls, children with active INS had higher percentages of Th2 cells ( < 0.05), but there was no significant difference in controls and children in remission. The plasma levels of IgE, IL-4, and IL-13 were significantly increased in children with active INS ( < 0.05). There were lower levels of miR-24 and miR-27 in children with active non-atopic INS ( < 0.05). Transfection experiments indicated that upregulation of each miRNA decreased the percentage of Th2 cells and the level of IL-4 ( < 0.05), and down-regulation of each miRNA had the opposite effects ( < 0.05). Children with active INS, with or without atopy, had higher levels of IgE, possibly related to their higher levels of IL-13 and IL-4 due to a drift toward Th2 cells. miR-24 and miR-27 suppressed the expression of Th2 cells and have a critical function regulating Th2 cell expression in INS.
我们通过测量两种特定的微小RNA（miRNA）对患有特发性肾病综合征（INS）儿童的Th2细胞的影响，来研究该疾病的发病机制。在获得知情同意后，我们招募了20名在开始使用类固醇之前处于活动期的INS儿童、20名在接受类固醇治疗后处于缓解期的INS儿童以及20名年龄匹配的健康对照。采用流式细胞术测量Th2细胞水平，采用细胞计数珠阵列测量免疫球蛋白E（IgE）、白细胞介素（IL）-4和IL-13水平。采用逆转录聚合酶链反应（RT-PCR）测量CD4TCD25细胞中miR-24和miR-27的水平。使用Ficoll密度梯度离心法分离外周血单个核细胞（PBMC），并用不同的模拟物或抑制剂miRNA进行转染。采用RT-PCR测量不同RNA的表达，采用流式细胞术确定Th2细胞的百分比。相对于健康对照，处于活动期的INS儿童的Th2细胞百分比更高（P<0.05），但对照组和缓解期儿童之间无显著差异。活动期INS儿童的血浆IgE、IL-4和IL-13水平显著升高（P<0.05）。活动期非特应性INS儿童的miR-24和miR-27水平较低（P<0.05）。转染实验表明，每种miRNA的上调均降低了Th2细胞的百分比和IL-4水平（P<0.05），而每种miRNA的下调则产生相反的效果（P<0.05）。有或无特应性的活动期INS儿童的IgE水平较高，这可能与其向Th2细胞漂移导致的IL-13和IL-4水平较高有关。miR-24和miR-27抑制Th2细胞的表达，并在INS中对调节Th2细胞表达起关键作用。
