Inhibitory effect of TGF-β1 on NO production in peritoneal macrophages from collagen-induced arthritis rats involving the LPS-TLR4 pathway

Wang,Ting-Yu; Li,Jun; Jin,Zhigui; Wu,Feihua; Zhou,Qian

doi:10.3892/mmr.2013.1641

Inhibitory effect of TGF-β1 on NO production in peritoneal macrophages from collagen-induced arthritis rats involving the LPS-TLR4 pathway

Authors:
- Ting-Yu Wang
- Jun Li
- Zhigui Jin
- Feihua Wu
- Qian Zhou
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Affiliations: Department of Pharmacy, Shanghai Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, P.R. China, School of Pharmacy, Anhui Medical University, Hefei, Anhui 230032, P.R. China
Published online on: August 16, 2013 https://doi.org/10.3892/mmr.2013.1641
Pages: 1143-1148

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Abstract

Transforming growth factor-β1 (TGF-β1) is critical in controlling inflammatory responses and the prevention of autoimmune diseases. Although the effect of TGF-β1 on macrophages from normal mice or rats has been established, little attention has been paid to its effect on disease conditions. In the present study, we investigated the regulatory effect, and possible mechanism, of TGF-β1 exposure on the secretion of nitric oxide (NO) in peritoneal macrophages (PMΦ) obtained from collagen-induced arthritis (CIA) rats. The CIA model was established by immunizing the emulsion of collagen type II and incomplete Freund's adjuvant (IFA) in Wistar rats. PMΦ were incubated with TGF-β1 (5 ng/ml) for 36 h and the supernatant, and cell mRNA and protein were collected. NO concentration was determined using an NO assay kit. The mRNA expression of inducible nitric oxide synthase (iNOS) and Toll-like receptor 4 (TLR4) was determined using reverse transcription-polymerase chain reaction (RT-PCR). The protein expression of iNOS was tested with western blot analysis. The expression of membrane TLR4 was determined by flow cytometry. We discovered that the secretion of NO from the PMΦ of CIA rats increased compared with normal rats. TGF-β1 significantly inhibited the production of NO in the PMΦ from CIA rats. iNOS mRNA and protein expression in the PMΦ from CIA rats may be suppressed by TGF-β1. TLR4 mRNA and protein expression in PMΦ from CIA rats were upregulated with LPS stimulation and treatment with TGF-β1 inhibited their expression. These results demonstrated that TGF-β1 inhibited lipopolysaccharide (LPS)-induced NO production in the PMΦ from CIA rats, which may be due to the inhibition of the LPS-TLR4 signaling pathway.

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