Expressions of Notch receptors and target gene in cultured fibroblast-like synoviocytes from osteoarthritis
WANG Hui1, HUA Shen-hao1, WANG Wen-hong2, WANG Xu-hui1,LIU Qiang1, CHEN Yan1,YOU Hai-yan1, JIAO Zhi-jun1
(1.Department of Laboratory Medicine, the Affiliated Hospital of Jiangsu University，Zhenjiang Jiangsu 212001；2.School of Medical Science and Laboratory Medicine，Jiangsu University，Zhenjiang Jiangsu 212013, China)
Abstract：Objective：To investigate the expression of Notch signaling molecules in fibroblast-like synoviocytes(FLS) from patients with osteoarthritis(OA). Methods: OA FLSs were isolated from synovial tissues and treated with LPS or LPS + DAPT between the passages of 3 and 5. RT-PCR was used to detect the expression of mRNA of the Notch receptors and Notch target gene Hes-1. In addition, the protein level of Hes-1 was detected by Western blot. Results: Compared with normal FLSs, the mRNA expression of Notch receptors 1, 2, 3 and Hes-1 was markedly increased in OA FLSs(P<0.05). LPS treatment significantly up-regulated the expression of Notch receptors1, 2, 3(Notch 4 level was low) and Hes-1. However, DAPT, a Notch signaling inhibitor, could down-regulated the protein level of Hes-1. Conclusion: Notch signaling was found to be activated in OA FLS, which may be involved in the pathophysiology of the inflammatory in OA joints.
[1]Ringdahl E, Pandit S. Treatment of knee osteoarthritis[J]. Am Fam Physician，2011， 83(11): 1287-1292.[2]de Lange-Brokaar BJ, Ioan-Facsinay A, van Osch GJ, et al. Synovial inflammation, immune cells and their cytokines in osteoarthritis: a review[J]. Osteoarthritis Cartilage， 2012， 20(12): 1484-1499.[3]Jiao Z, Wang W, Guo M, et al. Expression analysis of Notch-related molecules in peripheral blood T helper cells of patients with rheumatoid arthritis[J]. Scand J Rheumatol， 2010， 39(1): 26-32.[4]Jiao Z, Wang W, Ma J,et al. Notch signaling mediates TNF-alpha-induced IL-6 production in cultured fibroblast-like synoviocytes from rheumatoid arthritis[J]. Clin Dev Immunol， 2012，2012: 350209.[5]Nam Y, Sliz P, Song L, et al. Structural basis for cooperativity in recruitment of MAML coactivators to Notch transcription complexes[J]. Cell， 2006， 124(5): 973-983.[6]Hoyne GF. Notch signaling in the immune system[J]. J Leukoc Biol， 2003，74(6): 971-981.[7]Wu L, Huang X, Li L, et al. Insights on biology and pathology of HIF-1alpha/-2alpha, TGFbeta/BMP, Wnt/beta-catenin, and NFkappaB pathways in osteoarthritis[J]. Curr Pharm Des， 2012， 18(22): 3293-3312.[8]Philippe L, Alsaleh G, Suffert G, et al. TLR2 expression is regulated by microRNA miR-19 in rheumatoid fibroblast-like synoviocytes[J]. J Immunol， 2012， 188(1): 454-461.[9]Nakazawa M, Ishii H, Aono H, et al. Role of Notch-1 intracellular domain in activation of rheumatoid synoviocytes[J]. Arthritis Rheum， 2001， 44(7): 1545-1554.[10]Nakazawa M, Ishii H, Nakamura H, et al. NFkappaB2(p52) promoter activation via Notch signaling pathway in rheumatoid synoviocytes[J]. Int J Mol Med， 2001， 7(1): 31-35.[11]Ishii H, Nakazawa M, Yoshino S,et al. Expression of notch homologues in the synovium of rheumatoid arthritis and osteoarthritis patients[J]. Rheumatol Int， 2001， 21(1): 10-14.[12]Mahjoub M, Sassi N, Driss M, et al. Expression patterns of Notch receptors and their ligands in human osteoarthritic and healthy articular cartilage[J]. Tissue Cell， 2012， 44(3): 182-194.[13]Czerwinski A, Valenzuela F, Afonine P, et al. N-{N-[2-(3,5-Difluorophenyl)acetyl]-(S)-alanyl}-(S)-phenylglycine tert-butyl ester(DAPT): an inhibitor of gamma-secretase, revealing fine electronic and hydrogen-bonding features[J]. Acta Crystallogr C， 2010， 66(Pt 12): 585-588.
2013, Vol. 23 
