Decorin对人僵直膝关节滑膜囊成纤维样滑膜细胞增殖及蛋白合成的抑制作用

Inhibitory effect of decorin on the proliferation and protein synthesis of fibroblast-like synoviocytes of the stiff knee joint synovial membrane in vitro

[目的]探讨重组核心蛋白多糖(decorin)对培养的人僵直膝关节滑膜成纤维样滑膜细胞(fibroblast-like synoviocytes,FLS)的增殖和Ⅰ型胶原蛋白及mRNA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FLS,分别加入不同质量浓度(10μg/ml,0.1μg/ml)的Decorin;培养24、48、72h后用二甲基噻唑二苯基四唑溴盐(MTT)法测定FLS的增殖速度;用流式细胞仪检测FLS的周期和凋亡;用RTPCR法检测FLS合成Ⅰ型前胶原蛋白(PcⅠ)mRNA的含量,westem Blot检测Ⅰ型胶原蛋白的合成量,并与对照组比较。[结果]10μg/ml的Decorin可有效抑制滑膜FLS的增殖(P<0.05),明显增加FLS处于G0/G1期的百分比(P<0.05),并通过下调PcⅠmRNA的表达,在转录水平上抑制PcⅠ的合成;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FLS的增殖及PcⅠmRNA的表达和Ⅰ型胶原蛋白合成,在防治膝关节僵直中起一定的作用。

[Objective] To investigate the effects of decorin on the proliferation,collagen type I concentration,and mRNA expression of fibroblast- like synoviocytes（ FLSs） of the stiff knee joint synovial membrane in order to provide theory bases for the treatment of knee joint stiffness. [Methods] FLS samples were isolated from the stiff knee joint synovial membrane and cultured in vitro. The FLS samples were treated with decorin at concentrations of 0. 1 μg / ml and 10 μg / ml. After culturing for 24,48,and 72 h,the cell proliferation rates were measured through MTT colorimetric determination. Cell cycle distributions and apoptosis were analyzed by using a flow cytometer. The mRNA level of collagen type I was detected in semiquantitative reverse transcription polymerase chain reaction,while collagen type I concentration was measured by using Western blot and compared with a negative control group. [Results] The FLS proliferations were significantly inhibited,and a significant increase in the percent of FLS at the G1 phase was induced by 10 μg / ml decorin（ P 0. 05）,protein C inhibitor（ PCⅠ） mRNA expression,and PCⅠ synthesis were significantly decreased. FLS in late apoptosis were not found in the negative control and experimental groups. [Conclusion] Recombinant human decorin can inhibit the proliferation of and decrease in PC Ⅰ mRNA expression,and PCⅠ synthesis of FLS of the stiff knee joint synovial membrane in vitro. These results suggest that decorin potentially contributes to therapy for human knee stiffness.