中国修复重建外科杂志

中国修复重建外科杂志

S100 钙结合蛋白 B 在骨性关节炎软骨损伤修复中的作用及机制研究

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目的 探讨 S100 钙结合蛋白 B(S100B)在骨性关节炎(osteoarthritis,OA)软骨损伤修复中的作用及机制。 方法 取 20 只新西兰兔随机分为对照组和模型组,每组 10 只。模型组兔右膝关节制动法制备软骨损伤模型,对照组不作任何处理。4 周后采用 ELISA 法检测关节液 IL-1β、TNF-α 水平,实时荧光定量 PCR(real-time fluorescence quantitative PCR,qRT-PCR)和 Western blot 检测软骨组织 S100B、FGF-2、FGF 受体 1(FGF receptor 1,FGFR1)基因及蛋白表达。分离培养人滑膜成纤维细胞(synovial fibroblasts,SF),观察过表达、干扰 S100B 以及拮抗 FGFR1 对细胞 IL-1β 和 TNF-α 水平(ELISA 法)以及 FGF-2 和 FGFR1 基因(qRT-PCR 检测)和蛋白(Western blot 检测)表达的影响。 结果 ELISA 检测示模型组兔关节液中 IL-1β 和 TNF-α 表达水平均明显高于对照组(P<0.05);qRT-PCR 和 Western blot 检测示,模型组兔软骨组织 S100B、FGF-2、FGFR1 mRNA 和蛋白表达量均显著高于对照组(P<0.05)。过表达和干扰 S100 能够分别显著升高和降低脂多糖(lipopolysaccharides,LPS)诱导的 IL-1β 和 TNF-α 水平及 FGF-2 和 FGFR1 mRNA 和蛋白表达,差异均有统计学意义(P<0.05)。而拮抗 FGFR1 能够显著降低 LPS 诱导的 IL-1β 和 TNF-α 水平及 FGF-2 和 FGFR1 mRNA 和蛋白表达,差异均有统计学意义(P<0.05)。 结论 S100B 能够调节 SF 炎性反应并可能影响 OA 软骨损伤修复,其机制可能与激活 FGF-2/FGFR1 信号通路有关。

Objective To investigate the role and mechanism of S100 calcium binding protein B (S100B) in osteoarthritis (OA) cartilage damage repair. Methods Twenty New Zealand rabbits were randomly divided into control group and model group, with 10 rabbits in each group. Rabbits in the model group were injured by the right knee joint immobilization method to make the artilage injury model, while the control group did not deal with any injury. After 4 weeks, the levels of interleukin-1β (IL-1β) and tumor necrosis factor α (TNF-α) in synovial fluid were detected by ELISA method; the mRNA and protein expressions of S100B, fibroblast growth factor 2 (FGF-2), and FGF receptor 1 (FGFR1) in cartilage tissue were examined by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot assay. Human synovial fibroblasts (SF) were isolated and cultured in vitro. The effects of S100B overexpression and knockdown on the levels of IL-1β and TNF-α (ELISA method) and the expressions of FGF-2 and FGFR1 gene (qRT-PCR) and protein (Western blot) were observed. Moreover, the effects of FGFR1 knockdown in above S100 overexpression system on the levels of IL-1β and TNF-α (ELISA method) and the expressions of FGF-2 and FGFR1 gene (qRT-PCR) and protein (Western blot) were observed. Results ELISA detection showed that the expressions of IL-1β and TNF-α in the synovial fluid of the model group were significantly higher than those of the control group (P<0.05); qRT-PCR and Western blot detection showed that the mRNA and protein expressions of S100B, FGF-2, and FGFR1 in cartilage tissue were significantly higher than those of the control group (P<0.05). Overexpression and knockdown S100 could respectively significantly increase and decrease lipopolysaccharides (LPS) induced IL-1β and TNF-α levels elevation and the mRNA and protein expressions of FGF-2 and FGFR1 (P<0.05); whereas FGFR1 knockdown could significantly decrease LPS induced IL-1β and TNF-α levels elevation and the mRNA and protein expressions of FGF-2 and FGFR1 (P<0.05). Conclusion S100B protein can regulate the inflammatory response of SF and may affect the repair of cartilage damage in OA, and the mechanism may be related to the activation of FGF-2/FGFR1 signaling pathway.

关键词: S100 钙结合蛋白 B; 滑膜成纤维细胞; 骨性关节炎; 软骨损伤; FGF-2; FGF 受体 1

Key words: S100 calcium binding protein B; synovial fibroblasts; osteoarthritis; cartilage damage; fibroblast growth factor 2; fibroblast growth factor receptor 1

引用本文: 朱立帆, 周建新, 曾金才, 张晓剑, 沈鹏程, 翁峰标. S100 钙结合蛋白 B 在骨性关节炎软骨损伤修复中的作用及机制研究. 中国修复重建外科杂志, 2018, 32(11): 1429-1434. doi: 10.7507/1002-1892.201804060 复制

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1. Allen KD, Golightly YM. State of the evidence. Curr Opin Rheumatol, 2015, 27(3): 276-283.
2. Heijink A, Gomoll AH, Madry H, et al. Biomechanical considerations in the pathogenesis of osteoarthritis of the knee. Knee Surg Sports Traumatol Arthrosc, 2012, 20(3): 423-435.
3. Goldring MB, Otero M, Plumb DA, et al. Roles of inflammatory and anabolic cytokines in cartilage metabolism: signals and multiple effectors converge upon MMP-13 regulation in osteoarthritis. Eur Cell Mater, 2011, 21: 202-220.
4. Montaseri A, Busch F, Mobasheri A, et al. IGF-1 and PDGF-bb Suppress IL-1β-Induced Cartilage Degradation through Down-Regulation of NF-κB Signaling: Involvement of Src/PI-3K/AKT Pathway. PLoS One, 2011, 6(12): e28663.
5. Yan D, Chen D, Cool SM, et al. Fibroblast growth factor receptor 1 is principally responsible for fibroblast growth factor 2-induced catabolic activities in human articular chondrocytes. Arthritis Res Ther, 2011, 13(4): R130.
6. Davidson D, Blanc A, Filion D, et al. Fibroblast growth factor (FGF) 18 signals through FGF receptor 3 to promote chondrogenesis. J Biol Chem, 2005, 280(21): 20509-20515.
7. Zreiqat H, Belluoccio D, Smith MM, et al. S100A8 and S100A9 in experimental osteoarthritis. Arthritis Res Ther, 2010, 12(1): R16.
8. Choi IY, Karpus ON, Turner JD, et al. Stromal cell markers are differentially expressed in the synovial tissue of patients with early arthritis. PLoS One, 2017, 12(8): e0182751.
9. Abramson SB, Attur M. Developments in the scientific understanding of osteoarthritis. Arthritis Res Ther, 2009, 11(3): 227.
10. Yammani RR. S100 proteins in cartilage: Role in arthritis. Biochim Biophys Acta, 2012, 1822(4): 600-606.
11. Choi W, Kawanabe H, Sawa Y, et al. Effects of bFGF on suppression of collagen type I accumulation and scar tissue formation during wound healing after mucoperiosteal denudation of rat palate. Acta Odontol Scand, 2008, 66(1): 31-37.
12. Deng T, Huang S, Zhou S, et al. Cartilage regeneration using a novel gelatin-chondroitin-hyaluronan hybrid scaffold containing bFGF-impregnated microspheres. J Microencapsul, 2007, 24(2): 163-174.
13. Wang X, Manner PA, Horner A, et al. Regulation of MMP-13 expression by RUNX2 and FGF2 in osteoarthritic cartilage. Osteoarthritis Cartilage, 2004, 12(12): 963-973.
14. Schmal H, Zwingmann J, Fehrenbach M, et al. bFGF influences human articular chondrocyte differentiation. Cytotherapy, 2007, 9(2): 184-193.
15. Im HJ, Muddasani P, Natarajan V, et al. Basic fibroblast growth factor stimulates matrix metalloproteinase-13 via the molecular cross-talk between the mitogen-activated protein kinases and protein kinase Cdelta pathways in human adult articular chondrocytes. J Biol Chem, 2007, 282(15): 11110-11121.
16. Muddasani P, Norman JC, Ellman M, et al. Basic fibroblast growth factor activates the MAPK and NFkappaB pathways that converge on Elk-1 to control production of matrix metalloproteinase-13 by human adult articular chondrocytes. J Biol Chem, 2007, 282(43): 31409-31421.
17. Yan D, Chen D, Im HJ. Fibroblast growth factor-2 promotes catabolism via FGFR1-Ras-Raf-MEK1/2-ERK1/2 axis that coordinates with the PKCδ pathway in human articular chondrocytes. J Cell Biochem, 2012, 113(9): 2856-2865.