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Abstract
Purpose: We investigated the pathogenesis of idiopathic nephrotic syndrome (INS) by measuring the effects two specific miRNAs on Th2 cells in children with this disease.Methods: 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 CD4+TCD25− 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.Results: Relative to healthy controls, children with active INS had higher percentages of Th2 cells (P < 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 (P < 0.05). There were lower levels of miR-24 and miR-27 in children with active non-atopic INS (P < 0.05). Transfection experiments indicated that upregulation of each miRNA decreased the percentage of Th2 cells and the level of IL-4 (P < 0.05), and down-regulation of each miRNA had the opposite effects (P < 0.05).Conclusion: 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.
Highlights
Idiopathic nephrotic syndrome (INS) is the most common renal disease in children, and a main cause of chronic renal failure in children from China [1]
The results showed that the levels of miR-24 and miR-27 were significantly lower in the ANA group than in the Ctrl group
We further examined the relationship between miR-24, miR27, and Th2 cells by transfection of PBMCs isolated from healthy volunteers and INS patients with ANA using six different miRNAs: negative control (Ctrl-m); miR-24 mimic; miR-27 mimic; miRNA inhibitor negative control (Ctrl-i); miR-24 inhibitor; or miR-27 inhibitor miRNAs in Idiopathic Nephrotic Syndrome
Summary
Idiopathic nephrotic syndrome (INS) is the most common renal disease in children, and a main cause of chronic renal failure in children from China [1]. The clinical manifestations of INS are proteinuria, low level of plasma albumin, hyperlipidemia, and edema. The prevalence is greatest in preschool children who are 3–5 years-old. About 80–90% of patients with kidney disease who are under 10 years-old have minimal change disease (MCD), a common cause of INS [2]. The specific etiology and mechanism of INS in children remain unclear. Previous studies showed that miRNAs in Idiopathic Nephrotic Syndrome
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