【摘要】 [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。更多还原

【Abstract】 [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.更多还原