p53治疗白介素-1β诱导兔膝关节炎的实验研究

p53 reduces synovitis and arthritis in IL-1β induced knee arthritis model

目的:以腺病毒(Ad)为载体、p53为治疗基因,研究p53基因对人白介素(IL)-1β诱导兔膝关节炎的疗效和机制。方法:以Ad-LacZ为对照,将Ad-p53和Ad-LacZ分别注入6只人白介素1β(hIL-1β)诱导兔膝关节炎模型的膝关节。分析滑膜组织病理改变和关节炎的相关指标变化,检测滑膜组织细胞凋亡情况;培养人类风湿关节炎(RA)患者的滑膜成纤维细胞(FLS),用四氮唑蓝比色法(MTT)测定p53对滑膜细胞生长及增殖的影响,流式细胞仪检测细胞周期和凋亡情况,测定上清IL-6的水平。此外,逆转录多聚酶链反应(RT-PCR)和蛋白印迹(Western blot)方法评价腺病毒能否将治疗基因p53有效转移至滑膜组织。结果:Ad-p53感染滑膜成纤维细胞和滑膜组织后,治疗基因p53 mRNA和蛋白的表达明显上调。p53能显著减轻hIL-1β诱导兔膝关节炎的炎症,抑制滑膜细胞增生,减少炎症细胞浸润。p53不能诱导RA-FLS凋亡,但能显著下调促炎症细胞因子IL-6的产生。结论:抑癌基因p53很可能主要通过抑制促炎症因子的产生来减轻关节和滑膜炎症,是RA基因治疗的有效因子之一。

Objective：To address the efficacy of p53 on the therapy of IL-1β-induced rabbit knee arthritis and identify the mechanism.Methods：Adenovirus（Ad）-p53 and Ad-LacZ particles were individually injected into knee joint through patellar ligaments of human IL-1β-induced rabbit arthritis model,six each group.Analysis of arthritis and characteristics of histology was used to evaluate the efficacy as well as the identification of apoptosis.Human rheumatoid arthritis（RA）fibroblast-like synoviocytes（FLS）were cultured and infected by Ad-p53.MTT analysis and flow cytometry were used to evaluate the growth and proliferation,and ELISA to detect cytokine production.In addition,the abilities of adenoviral vectors to transfer p53 gene to synovium and synoviocytes were assessed with RT-PCR and Western blot.Results：Delivery of p53 mediated by adenovirus was successful in FLS and synovium with up-regulation of p53 mRNA and p53 protein.P53 significantly improved synovitis and arthritis of hIL-1β-induced lapine arthritis.Infiltration of inflamed cells was decreased.But apoptosis was not detected in FLS and synovium as expected.Interestingly,proinflammatory cytokine IL-6 was significantly suppressed by p53.Conclusion：Tumor suppressor,p53 might improve synovitis and arthritis mainly through the inhibition of proinflammatory cytokines production in FLS,and will be a novel target for the therapy on RA.