Abstract | INTRODUCTION: METHODS:
Arthritis in DA rats was induced by pristane and the severity was evaluated by macroscopic and microscopic score systems. Spleen TLR and cytokine expression was detected at different time points by real-time polymerase chain reaction (PCR) and flow cytometry. Polyinosine- polycytidylic acid (polyI:C, a ligand of TLR3) or TLR3 specific short-hairpin RNA plasmid for RNA interference was administrated to PIA rats in vivo. Serum nitrogen oxide concentration was determined by Griess method, and tumor necrosis factor alpha ( TNF-alpha) was determined by L929 biotest. In splenic macrophages, TLR3 expression was measured by flow cytometry. A rat macrophage cell line (NR8383) was stimulated by pristane, and anti-TLR3 antibody were used to block TLR3 pathway. TLR3 and cytokine expression in NR8383 were detected by real-time PCR. RESULTS: By screening the TLR expression profile in spleen of DA rats after pristane injection, we found that TLR3 was the most early and prominently upregulated TLR. Both TLR3 mRNA and protein expression of spleen were upregulated at 6 and 26 days after pristane injection. Furthermore, administration of polyI:C exacerbated, whereas RNA interference targeting TLR3 ameliorated, the arthritis. Particularly, TLR3 expression was induced in splenic macrophages of PIA rats, and also in the NR8383 cell line after pristane stimulation in a dose- and time- dependent manner. Upregulation of interferon beta (IFN-beta) and TNF-alpha by pristane stimulation was blocked by anti-TLR3 antibody in NR8383. CONCLUSIONS: TLR3 plays a pivotal role in the initiation and development of PIA which may dependent on macrophage. These findings are useful to understand the pathogenesis of RA and may provide an intriguing therapeutic opportunity for RA.
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Authors | Liesu Meng, Wenhua Zhu, Congshan Jiang, Xiaojing He, Weikun Hou, Fang Zheng, Rikard Holmdahl, Shemin Lu |
Journal | Arthritis research & therapy
(Arthritis Res Ther)
Vol. 12
Issue 3
Pg. R103
( 2010)
ISSN: 1478-6362 [Electronic] England |
PMID | 20500834
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- TLR3 protein, rat
- Terpenes
- Tlr9 protein, rat
- Toll-Like Receptor 3
- Toll-Like Receptor 9
- pristane
- Poly I-C
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Topics |
- Animals
- Arthritis, Experimental
(chemically induced, metabolism, pathology)
- Cell Line
- Disease Models, Animal
- Macrophages
(metabolism, pathology)
- Poly I-C
(pharmacology)
- RNA Interference
- Rats
- Rats, Inbred Strains
- Severity of Illness Index
- Spleen
(metabolism)
- Terpenes
(adverse effects)
- Toll-Like Receptor 3
(genetics, metabolism)
- Toll-Like Receptor 9
(metabolism)
- Up-Regulation
(drug effects, physiology)
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