中文摘要：

目的：通过TIPE2表达质粒转染大鼠佐剂型关节炎 （AA） 成纤维样滑膜细胞 （FLS）,体外研究 AA 的发病中 TIPE2 对DR5介导细胞凋亡的作用.方法：应用 MIGR1/TIPE2＋/＋ 表达质粒,经脂质体法导入AA-FLS细胞中,转染72小时后,荧光显微镜检测质粒自带GFP基因的表达情况确认转染效率,以未作处理的AA-FLS为对照组,用半定量逆转录聚合酶链反应 （ RT-PCR ） 检测 TIPE2 mRNA的表达,用 Western blot 法检测TIPE2 蛋白的表达变化,MTT 法以及流式细胞术检测抗 DR5 功能性单链抗体ZF1对两组细胞生长的影响.结果：成功获得荧光强度90% 的 TIPE2过表达 AA-FLS 细胞,TIPE2 mRNA 及蛋白表达显著提高.抗 DR5 单链抗体 ZF1 对 TIPE2＋/＋ 的FLS组细胞具有明显生长抑制及凋亡诱导作用,与不作处理的FLS组相比差异显著.结论：TIPE2＋/＋表达质粒明显提升AA-FLS的TIPE2蛋白表达水平,TIPE2 对 DR5 介导细胞凋亡有重要作用.

英文摘要：

Objective： Fibroblast-like synoviocytes from adjuvant arthritis （AA） rat were transfected with plasmid over-ex- pressed TIPE2＋/＋ , then apoptosis mediated by TIPE2 on DR5 and its pathological role in adjuvant arthritis （AA） were studied in vitro. Methods： The plasmid of MIGR1/TIPE2 ＋/＋ were transfected to AA-FLS by liposome, after 72 h, the expression of GFP gene that plasmid owned was identified by fluorescence microscope in confirm transfection efficiency, TIPE2 mRNA expression was detected by semi-quantitative reverse transcription polymerase chain reaction （ RT-PCR）, TIPE2 protein expression of AA-FLS were detected by Western blot. The growth resistance of cells mediated by ZF1 （ DR5 single chain antibody） was detected by MTF method and flow cy- tometry. Results： Plasmid over-expressed TIPE2 ＋/＋ were transfected to AA-FLS successfully as the fluorescence intensity reached 90% ,TIPE2 mRNA and protein expression were significantly improved. Furthermore, the growth inhibition and apoptosis induction effect on FLS/TIPE2 ＋/＋ were significant, compared with FLS without processing. Conclusion： TIPE2 expression level in AA-FLS are significantly improved by transfecting plasmid expressed TIPE2 ＋/＋ , TIPE2 plays an important pathological role in apoptosis mediated by DRS.