重组人内抑素对佐剂性关节炎大鼠成纤维样滑膜细胞周期和PCNA表达的影响

Effects of recombinant human endostatin on cell cycle and PCNA expression in fibroblast-like synoviocytes in rats with adjuvant arthritis

目的观察重组人内抑素(rhEndostatin)对佐剂性关节炎(AA)大鼠成纤维样滑膜细胞(FLS)细胞周期及增殖细胞核抗原(PCNA)表达的影响,探讨rhEndostatin抑制AAFLS增殖的分子机制。方法制备AA大鼠模型,应用流式细胞仪分析rhEndostatin对AA FLS细胞周期的影响;分别采用实时荧光定量PCR和Western blot方法,定量分析rhEn-dostatin对AA大鼠滑膜组织PCNA mRNA及蛋白表达的影响。结果与正常组相比,AA FLS G1期细胞减少(P<0.01),S期和G2/M期细胞增加(P<0.01);AA大鼠滑膜组织PCNA mRNA和蛋白表达增加(P<0.05,P<0.01)。与AA模型组相比,rhEndostatin治疗组细胞G1期比例增加(P<0.01),S期和G2/M期细胞比例减少(P<0.01);滑膜组织PCNA mRNA和蛋白表达减少(P<0.01)。结论rhEn-dostatin可引起AA FLS细胞周期G1期阻滞,该作用可能与其抑制AA FLS PCNA表达有关。

Aim To observe the effects of recombinant human endostatin（rhEndostatin） on cell cycle and the expression of proliferating cell nuclear antigen （PC- NA） in fibroblast-like synoviocytes in rats with adjuvant arthritis （AA）, and to explore the molecular mechanisms of the inhibitory effect of rhEndostatin on proliferation of fibroblast-like synoviocytes （ FLS ） in AA rats. Methods Adjuvant arthritis was induced by Freund＇s complete adjuvant in rats. Flow cytometry was applied to measure the effects of rhEndostatin on the cell cycle in AA FLS. The effect of rhEndostatin on the expression of PCNA mRNA and protein in synovial tissue in AA rats was examined quantitatively by realtime fluorescent quantitative PCR and Western blot assays, respectively. Results The percentage of cells in G1 phase in AA FLS decreased significantly （P 〈 0. 01 ） , and the rate of cells in S and GE/M phase increased significantly（P 〈 0.01 ） , compared with that innormal controls. The expression levels of PCNA mRNA and protein in AA synovial tissue were significantly higher than those in normal samples （ P 〈 0. 05, P 〈 0. 01）. However, the percentage of cells in Gl phase in FLS treated with rhEndostatin increased significantly （P 〈0.01）, and the rate of cells in S and G2/M phase decreased significantly （ P 〈 0. 01 ）, compared with that in AA controls. The expression levels of PC-NA mRNA and protein in synovial tissue treated with rhEndostatin were significantly lower than those in AA group （ P 〈 0.01 ）. Conclusion Recombinant human endostatin causes cell cycle arrest in G1 in AA FLS, which may be involved in its inhibitory effect on PCNA expression in AA FLS.