Decorin对僵直膝关节滑膜B型细胞增殖及I型胶原蛋白表达的影响

目的:探讨重组核心蛋白聚糖(decorin)对人僵直膝关节滑膜B型细胞的增殖和Ⅰ型前胶原蛋白(PcⅠ)mRNA表达及Ⅰ型胶原蛋白(ColⅠ)合成的影响,为decorin治疗人膝关节僵直提供理论依据。方法:分离培养人僵直膝关节滑膜B型细胞,分别加入不同浓度(0.1、5和10 mg/L)的decorin;培养24 h、48 h和72 h后用MTT法测定关节滑膜B型细胞的活力;用流式细胞术检测滑膜B型细胞的周期和凋亡;RT-PCR法检测滑膜B型细胞的PcⅠmRNA表达;Western blot检测ColⅠ的合成。结果:5和10 mg/L的decorin可有效降低滑膜B型细胞的活力(P0.05),明显增加处于G0/G1期细胞的百分比(P0.05),并下调PcⅠmRNA的表达,抑制ColⅠ的合成;同时实验组和对照组均未检测到终末期凋亡细胞。结论:Decorin可抑制人僵直膝关节滑膜B型细胞的增殖、PcⅠmRNA的表达和ColⅠ的合成,在防治膝关节僵直中可能起一定的作用。

Effect of decorin on proliferation and collagen type I synthesis of stiff knee joint synovial type B cells

AIM: To explore the effects of decorin on procollagen type I (PcI), mRNA expression,collagen type I synthesis and proliferation of synovial type B cells of stiff knee joint synovial membrane. METHODS: Type B cells of synovial membrane were isolated from the stiff knee joint synovial membrane and cultured in vitro. The cells were treated with decorin at concentrations of 0.1 mg/L, 5 mg/L and 10 mg/L. After cultured for 24 h, 48 h and 72 h, the cell proli-feration rates were measured by MTT colorimetric determination. Cell cycle distribution and apoptosis were analyzed by flow cytometry. The mRNA level of Pc I was detected by RT-PCR, while collagen type I was measured by Western blot. RESULTS: The proliferation of synovial type B cells was significantly inhibited, the percentage of synovial type B cells at G₁ phase was significantly increased by 5 mg/L and 10 mg/L decorin (P<0.05), and PcⅠmRNA expression and collagen type I synthesis were significantly decreased. The cells with late apoptosis were not found in control group and experimental groups. CONCLUSION: Recombinant human decorin inhibits synovial type B cell proliferation and decreases PcⅠmRNA expression and collagen type I synthesis in synovial type B cells of stiff knee joint synovial membrane in vitro, suggesting that decorin potentially contributes to the therapy of human knee stiffness.