TRIM28/TIF1β and Fli-1 negatively regulate peroxynitrite generation via DUOX2 to decrease the shedding of membrane-bound fractalkine in human macrophages after exposure to substance P

被引：3

作者：

Yamaguchi, Rui ^{[1
]}

Haraguchi, Misa ^{[1
]}

Yamaguchi, Reona ^{[2
,3
]}

Sakamoto, Arisa ^{[1
]}

Narahara, Shinji ^{[1
]}

Sugiuchi, Hiroyuki ^{[1
]}

Yamaguchi, Yasuo ^{[1
]}

机构：

[1] Kumamoto Hlth Sci Univ, Grad Sch Med Sci, Kitaku Izumi Machi 325, Kumamoto 8615598, Japan

[2] Kyoto Univ, Grad Sch Med, Dept Neurosci, Sakyo Ku, Yoshida Konoe Cho, Kyoto 6068501, Japan

[3] Kyoto Univ, Fac Med, Sakyo Ku, Yoshida Konoe Cho, Kyoto 6068501, Japan

来源：

CYTOKINE | 2020年 / 134卷

关键词：

DUOX2; Fractalkine; ADAM family; Peroxynitrite; Shedding; NITRIC-OXIDE SYNTHASE; ALPHA-CONVERTING ENZYME; NECROSIS-FACTOR-ALPHA; FACTOR-DEPENDENT MACROPHAGES; TGF-BETA; RHEUMATOID-ARTHRITIS; GENE-EXPRESSION; E3; LIGASE; RELEASE; PHOSPHATIDYLSERINE;

D O I：

10.1016/j.cyto.2020.155180

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The chemokine fractalkine is synthesized as a membrane-bound protein, but studies have shown that serum levels of soluble fractalkine are elevated in inflammatory and autoimmune diseases. Patients with autoimmune diseases also have increased serum levels of neuropeptide substance P (SP). The shedding activity of the ADAM family is induced by peroxynitrite, but that of SP is unclear. Treatment of human macrophages with SP upregulated levels of membrane-bound fractalkine. Interestingly, small interfering RNA (siRNA) for DUOX2 further increased membrane-bound fractalkine but decreased soluble fractalkine compared with cells treated with SP alone. SP induced nitric oxide 2/inducible nitric oxide synthase (NOS2/iNOS) mRNA and increased levels of nitrotyrosine, a biomarker of peroxynitrite, whereas transfection with DUOX2 siRNA blunted upregulation of nitrotyrosine. Most importantly, N(w)-nitro-L-arginine methyl ester (L-NAME, a nitric oxide synthase inhibitor) decreased protein levels of nitrotyrosine and concomitantly increased expression of membrane-bound fractalkine after exposure to SP. As for the signaling pathway of TGF beta 1 (an inhibitor of iNOS mRNA expression), silencing of RNA for TAK-1 upregulated membrane-bound fractalkine, but silencing of RNA for the Smad family did not. Interfering RNA of transcription factor specificity protein 1 (Sp1) upregulated protein levels of TGF beta 1/LAP. Most importantly, double transfection with siRNA for Sp1 and TRIM28/TIF1 beta or Fli-1 led to a significant increase in TGF beta 1/LAP levels and a corresponding reduction of NOS2/iNOS, which inhibited the shedding of membrane-bound fractalkine. In conclusion, TRIM28/TIF1 beta and Fli-1 negatively regulate TGF beta 1 expression to upregulate the generation of peroxynitrite, leading to increased shedding of membrane-bound fractalkine induced by SP.

引用

页数：9