维甲酸诱导基因-I 由双链 RNA 诱导，并调节人肾小球系膜细胞中 CC 趋化因子配体（CCL）5 的表达。

Retinoic acid-inducible gene-I is induced by double-stranded RNA and regulates the expression of CC chemokine ligand (CCL) 5 in human mesangial cells.

机构信息

Department Vascular Biology, Hirosaki University Graduate School of Medicine, Zaifu-cho, Hirosaki, Japan.

出版信息

Nephrol Dial Transplant. 2010 Nov;25(11):3534-9. doi: 10.1093/ndt/gfq270. Epub 2010 May 19.

DOI:10.1093/ndt/gfq270

PMID:20484300

Abstract

BACKGROUND

Retinoic acid-inducible gene-I (RIG-I) is a putative RNA helicase involved in immune reactions against RNA viruses and various inflammatory and autoimmune diseases. The purpose of the present study was to investigate the role of RIG-I in glomerular diseases.

METHODS

We treated human mesangial cells in culture with polyinosinic-polycytidylic acid (poly IC), which is an authentic double-stranded RNA, and analysed the expression of RIG-I, CC chemokine ligand 5 (CCL5) and interferon (IFN)-β by western blotting, reverse transcriptase-polymerase chain reaction (RT-PCR) or enzyme-linked immunosorbent assay (ELISA). To elucidate the poly IC-signalling pathway, we subjected the cells to RNA interference (RNAi) against RIG-I, IFN-β or Toll-like receptor (TLR) 3. Furthermore, we studied the effects of IFN-β receptor blocking and IFN-β overexpression.

RESULTS

Poly IC induced the expression of RIG-I and CCL5 in human mesangial cells, and RNAi against RIG-I inhibited this poly IC-induced CCL5 expression. Poly IC-induced RIG-I expression was also inhibited by RNAi against IFN-β and by an antibody against the IFN-β receptor. IFN-β overexpression induced the expression of both RIG-I and CCL5. The knockdown of TLR3 abolished poly IC-induced RIG-I expression.

CONCLUSIONS

The TLR3/IFN-β/RIG-I/CCL5 signalling pathway may mediate immune and inflammatory responses against viral infection in mesangial cells, suggesting the role of this pathway in the aggravation of glomerulonephritis due to viral infection.

摘要

背景

维甲酸诱导基因-I（RIG-I）是一种假定的 RNA 解旋酶，参与针对 RNA 病毒和各种炎症及自身免疫性疾病的免疫反应。本研究旨在探讨 RIG-I 在肾小球疾病中的作用。

方法

我们用 poly IC（一种真实的双链 RNA）处理培养的人系膜细胞，通过 Western blot、逆转录-聚合酶链反应（RT-PCR）或酶联免疫吸附试验（ELISA）分析 RIG-I、CC 趋化因子配体 5（CCL5）和干扰素（IFN）-β的表达。为阐明 poly IC 信号通路，我们用 RIG-I、IFN-β 或 Toll 样受体（TLR）3 的 RNA 干扰（RNAi）处理细胞。此外，我们还研究了 IFN-β 受体阻断和 IFN-β 过表达的影响。

结果

poly IC 诱导人系膜细胞中 RIG-I 和 CCL5 的表达，用 RIG-I 的 RNAi 可抑制这种 poly IC 诱导的 CCL5 表达。IFN-β 的 RNAi 和 IFN-β 受体的抗体也抑制了 poly IC 诱导的 RIG-I 表达。IFN-β 过表达诱导 RIG-I 和 CCL5 的表达。TLR3 的敲低消除了 poly IC 诱导的 RIG-I 表达。