Effects of fractalkine on nuclear factor-κB activation and endogenous fractalkine mRNA expression in fibroblast-like synoviocytes from patients with rheumatoid arthritis |
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Authors: | GUO Xing-hua PAN Yun-feng SONG Ze-rong WANG Kun GU Jie-ruo |
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Institution: | 1. Department of Rheumatology, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510630, China;
2. Department of Orthopedics, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510630, China |
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Abstract: | AIM: To investigate the effects of fractalkine(FKN) on nuclear factor kappa B (NF-κB) activation and endogenous FKN mRNA expression in fibroblast-like synoviocytes (FLS) from the patients with rheumatoid arthritis (RA). METHODS: RA-FLS were gained through tissue culture. Fractalkine at 100 μg/L was used to stimulate RA-FLS for 0 h, 1 h and 2 h. The expression of NF-κB p65 protein in cytoplasm and nucleus was detected by Western blotting, representing the activation of NF-κB in RA-FLS. RA-FLS was stimulated with fractalkine at concentration of 100 μg/L for 0 h, 12 h or 18 h, and the mRNA expression of FKN in RA-FLS was detected by RT-PCR.RESULTS: After stimulated with recombinant human FKN for 1 h, the expression of NF-κB p65 protein in the cytoplasm of RA-FLS was obviously lower than that in RA-FLS without FKN treatment in control group (P<0.05). After stimulated with FKN for 2 h, the expression of NF-κBp65 protein in nucleus was obviously higher than that in RA-FLS of control group (P<0.05). Recombinant human FKN at concentration of 100 μg/L induced endogenous FKN mRNA expression in RA-FLS in a time-dependent manner. The mRNA expression of FKN in RA-FLS obviously increased after stimulated with FKN for 18 h (P<0.05). CONCLUSION: FKN up-regulates the expression of endogenous FKN mRNA, suggesting a positive feedback. FKN can activate the NF-κB and may play an important role in the beginning of joint inflammation, angiogenesis and bone destruction. |
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Keywords: | Fractalkine Arthritis rheumatoid Fibroblast-like synoviocytes NF-kappa B |
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