程序化细胞死亡因子5过表达促进雷公藤内醇酯诱导的类风湿关节炎成纤维样滑膜细胞凋亡

Overexpression of progrmmed cell death 5 factor enhances triptolides-induced fibroblast-like synoviocytes apoptosis in rheumatoid arthritis

目的:探讨程序化细胞死亡因子5(progrmmed cell death5,PDCD5)过表达对雷公藤内醇酯(triptolide)诱导类风湿关节炎成纤维样滑膜细胞(rheumatiod arthritis fibroblast-likes synoviocytes,RAFLS)凋亡的作用。方法:体外分离培养RAFLS进行腺病毒Ad-PDCD5(含有PDCD5基因的腺病毒载体)的转染,分别用免疫印迹法和流式细胞术检测Ad-PDCD5转染后的RAFLS中PDCD5蛋白表达及经雷公藤内醇酯处理后的PDCD5蛋白过表达的RAFLS的凋亡率。结果:用感染复数(multiplicity of infection,MOI)分别为50、100、200和300的Ad-PDCD5感染RAFLS36h后,PDCD5蛋白表达水平呈现剂量依赖性增加。未经雷公藤内醇酯处理的非感染组、Ad-null组(不含目的基因的空载体)和Ad-PDCD5组RAFLS的凋亡之间差异无统计学意义,经雷公藤内醇酯处理后,其RAFLS的凋亡率分别为(22.51±3.87),(28.77±12.97)和(48.87±12.69)。表明此腺病毒体系能够有效地使PDCD5蛋白过表达,在RAFLS中,单独使用雷公藤内醇酯或者单独转染PDCD5都不能明显地诱导细胞凋亡,但过表达PDCD5再进行雷公藤内醇酯处理可以诱导细胞凋亡。结论:腺病毒Ad-PDCD5转染使PDCD5过表达,从而能够促进雷公藤内醇酯诱导的RAFLS凋亡,为类风湿关节炎的临床治疗提供了一个可能的干预靶点。

Objective： To explore the effect of prognnmed cell death 5 （ PDCD5 ） on apoptosis of rheumatoid arthritis fibroblast-like synoviocytes（ RA FLS） induced by triptolide. Methods： Cultured synovial ceils in vitro from RA patients were transfected with Ad-PDCD5. At protein level, expression of PDCD5 in RA FLS infected with Ad-PDCD5 was detected by Western blot. RA FLS infected with Ad-PDCD5 were cultured in presence or absence of triptolide and apoptosis of RA FLS was determined by flow cytometry. Results： Infection of RA FLS with increasing concentrations of Ad-PDCD5 （50 -300 MOI） resulted in a does-dependent increase in the production of PDCD5. Apoptotic cells percentage for noinfection group, Adull group and Ad-PDCD5 group were （ 22.41 ± 3. 87 ） %, （ 28.77 ± 12. 97 ） % and （ 48.87 ± 12. 69）%, respectively. Alternatively, infection without addition of triptolide stimuli had no effect. The data showed that gene transfection of PDCD5 alone without addition of triptolide was not sufficient to activate RA FLS apoptosis, PDCD5 acted as an enhancer rather than inductor of apoptosis. Conclusion： Overexpression of PDCD5 could enhance apoptosis of RA FLS induced by triptolide, PDCD5 may be a potential therapeutic target to RA.