miR-499 通过靶向钙调神经磷酸酶减轻微小病变性肾病足细胞损伤。

miR-499 Ameliorates Podocyte Injury by Targeting Calcineurin in Minimal Change Disease.

出版信息

Am J Nephrol. 2018;47(2):94-102. doi: 10.1159/000486967. Epub 2018 Feb 15.

Abstract

BACKGROUND

Podocyte injury is a hallmark of minimal change disease (MCD). Calcineurin inhibitors have been widely used in the current treatment of MCD, and miR-499 may target calcineurin. We aimed to study the function of miR-499 in MCD and test whether miR-499 delivery can improve MCD.

METHODS

An MCD mouse model was generated using puromycin aminonucleoside (PAN). MiR-499 was delivered using lentiviruses. Biochemical indicators including serum albumin, triglyceride, cholesterol, and 24-h urine protein were determined. Targets of miR-499 were confirmed using reporter gene activity assays. The ultrastructure of podocytes was analyzed using transmission electron microscopy.

RESULTS

MiR-499 significantly improved MCD-related symptoms and signs. Foot-process effacement was caused by PAN and partially reversed by miR-499. We identified that both CnAα and CnAβ were targets of miR-499, and were overexpressed in the presence of PAN. However, miR-499 reduced the expression of CnAα and CnAβ, leading to a decreased activity of calcineurin signaling in mouse podocytes in vitro and in vivo. In addition, miR-499 recovered PAN-induced reduction of cell viability.

CONCLUSIONS

MiR-499 ameliorated podocyte injury by targeting CnAα and CnAβ in a PAN-induced MCD mouse model. Delivery of miR-499 can be a novel strategy for MCD treatment.

摘要

背景

足细胞损伤是微小病变性肾病（MCD）的标志。钙调神经磷酸酶抑制剂已广泛用于 MCD 的当前治疗，miR-499 可能是钙调神经磷酸酶的靶点。我们旨在研究 miR-499 在 MCD 中的作用，并测试 miR-499 传递是否可以改善 MCD。

方法

使用嘌呤霉素氨基核苷（PAN）生成 MCD 小鼠模型。使用慢病毒传递 miR-499。测定血清白蛋白、甘油三酯、胆固醇和 24 小时尿蛋白等生化指标。使用报告基因活性测定法确认 miR-499 的靶标。使用透射电子显微镜分析足细胞的超微结构。

结果

miR-499 显著改善了与 MCD 相关的症状和体征。足突融合是由 PAN 引起的，并部分被 miR-499 逆转。我们发现 CnAα 和 CnAβ 都是 miR-499 的靶标，并且在存在 PAN 的情况下表达过度。然而，miR-499 降低了 CnAα 和 CnAβ 的表达，导致体外和体内鼠足细胞中钙调神经磷酸酶信号通路的活性降低。此外，miR-499 恢复了 PAN 诱导的细胞活力降低。