TLR4 signaling induces retinoic acid-inducible gene-I and melanoma differentiation-associated gene 5 in mesangial cells

Authors:
Imaizumi T1, Aizawa-Yashiro T, Watanabe S, Matsumiya T, Yoshida H, Tatsuta T, Xing F, Meng P, Hayakari R, Tsuruga K, Tanaka H
In:
Source: Other
Publication Date: (2013)
Issue: 26(5): 886-93
Cells used in publication:
Mesangial cells, Human (NHMC)
Species: human
Tissue Origin: kidney
Abstract
BACKGROUND: It is known that recognition of bacterial lipopolysaccharide (LPS) and various endogenous ligands by Toll-like receptor 4 (TLR4) induces inflammatory reactions. However, the role of TLR4 activation in mesangial inflammation remains to be elucidated. Retinoic acid-inducible gene-I (RIG-I) and melanoma differentiation-associated gene 5 (MDA5) are putative RNA helicases and are involved in immune and inflammatory reactions. The purpose of the present study was to investigate the implication of RIG-I and MDA5 in TLR4 signaling in mesangial cells. METHODS: Normal human mesangial cells in culture were treated with LPS. Expression of RIG-I, MDA5, interferon-ß (IFN-ß), CXCL10 and CXCL8 was examined using real-time RT-PCR, Western blotting and ELISA. The cells were also subjected to RNA interference against TLR4, IFN-ß, RIG-I or MDA5. RESULTS: LPS induced the expression of IFN-ß, RIG-I, MDA5, CXCL8 and CXCL10 in human mesangial cells. RNA interference against either TLR4 or IFN-ß inhibited LPS-induced RIG-I and MDA5 expression. Knockdown of TLR4, IFN-ß, RIG-I or MDA5 resulted in decreased induction of CXCL10, while only TLR4 knockdown inhibited CXCL8 induction. CONCLUSIONS: TLR4 signaling induces the expression of RIG-I and MDA5 in mesangial cells. RIG-I and MDA5 may be involved in inflammatory reactions by regulating CXCL10 expression in the downstream of TLR4 signaling in human mesangial cells.