Effect of Sinomenine on Proliferation of Fibroblast-like Synoviocytes and Expression of α7nAChR,TLR2 and TLR4
Author(s): PENG Chong, YI Lang, LYU Yanjun, BAI Shasha, LI Jing, DONG Yan, WANG Peixun, Department of Immunology, Guangzhou University of Chinese Medicine
Pages: 503-508
Year: 2016 Issue: 4
Journal: Traditional Chinese Drug Research & Clinical Pharmacology
Keyword: sinomenine; FLS; α7nAChR; TLR2; TLR4; proliferation;
Abstract: Objective To study the effect of sinomenine（SIN）on the proliferation of rat fibroblast-like synoviocytes（FLS）stimulated by tumor necrosis factor-α（TNF-α）and on the expression of α7 nicotinic acetylcholine receptors（α7n ACh R）,toll-like receptor 2（TLR2）and TLR4 in rats. Methods Primary FLS were cultured using tissue explant method. And then CD90,the surface marker of FLS,was measured by flow cytometry. FLS were divided into blank control group,TNF-α group and SIN groups(the concentration being 100,200,400 μmol·L-1,respectively). SIN groups were pretreated with SIN for 30 min,and then TNF-α group and SIN groups were stimulated by 20 ng·m L-1TNF-α for 24 h. The proliferation of FLS of each group was measured by MTT assay,the release of serum interleukin6（IL-6） was measured by enzyme-linked immunosorbent assay（ELISA）,and the m RNA expression of α7n ACh R,TLR2 and TLR4 was measured by RT-PCR and their protein expression was detected by Western blot method. Results After stimulating by TNF-α, the proliferation of FLS was significantly increased, and the release of IL-6 in the supernatant was also increased distinctly. Furthermore,the m RNA and protein expression of α7n ACh R,TLR2 and TLR4 were upregulated. However,the proliferation of FLS and the release of supernatants IL-6 were counteracted,and the expression of m RNA and protein of α7n ACh R,TLR2 and TLR4 were downregulated by SIN at 200,400μmol·L-1. Conclusion The proliferation of FLS is related with the expression of α7n ACh R,TLR2 and TLR4. SIN can inhibit the proliferation of FLS,and downregulate the expression of α7n ACh R,TLR2 and TLR4.
Pages: 503-508
Year: 2016 Issue: 4
Journal: Traditional Chinese Drug Research & Clinical Pharmacology
Keyword: sinomenine; FLS; α7nAChR; TLR2; TLR4; proliferation;
Abstract: Objective To study the effect of sinomenine（SIN）on the proliferation of rat fibroblast-like synoviocytes（FLS）stimulated by tumor necrosis factor-α（TNF-α）and on the expression of α7 nicotinic acetylcholine receptors（α7n ACh R）,toll-like receptor 2（TLR2）and TLR4 in rats. Methods Primary FLS were cultured using tissue explant method. And then CD90,the surface marker of FLS,was measured by flow cytometry. FLS were divided into blank control group,TNF-α group and SIN groups(the concentration being 100,200,400 μmol·L-1,respectively). SIN groups were pretreated with SIN for 30 min,and then TNF-α group and SIN groups were stimulated by 20 ng·m L-1TNF-α for 24 h. The proliferation of FLS of each group was measured by MTT assay,the release of serum interleukin6（IL-6） was measured by enzyme-linked immunosorbent assay（ELISA）,and the m RNA expression of α7n ACh R,TLR2 and TLR4 was measured by RT-PCR and their protein expression was detected by Western blot method. Results After stimulating by TNF-α, the proliferation of FLS was significantly increased, and the release of IL-6 in the supernatant was also increased distinctly. Furthermore,the m RNA and protein expression of α7n ACh R,TLR2 and TLR4 were upregulated. However,the proliferation of FLS and the release of supernatants IL-6 were counteracted,and the expression of m RNA and protein of α7n ACh R,TLR2 and TLR4 were downregulated by SIN at 200,400μmol·L-1. Conclusion The proliferation of FLS is related with the expression of α7n ACh R,TLR2 and TLR4. SIN can inhibit the proliferation of FLS,and downregulate the expression of α7n ACh R,TLR2 and TLR4.
Related Articles