程序性细胞死亡因子5增强雷公藤甲素诱导的类风湿关节炎成纤维样滑膜细胞凋亡

Programmed cell death 5 factor enhances triptolide-induced fibroblast-like synoviocyte apoptosis of rheumatoid arthritis.

作者信息

Jiang Jun, Wang Ning, Guan Zhenpeng, Houshan L V

机构信息

Arthritis Clinical & Research Center, Peking University People's Hospital, Beijing, China.

出版信息

Artif Cells Blood Substit Immobil Biotechnol. 2010;38(1):38-42. doi: 10.3109/10731190903495769.

DOI:10.3109/10731190903495769

PMID:20047520

Abstract

OBJECTIVE

To study the effect of programmed cell death 5 (PDCD5) on apoptosis of rheumatoid arthritis fibroblast-like synoviocyte (RAFLS) induced by triptolide.

METHOD

Cultured synovial cells in vitro from RA patients were transfected with Ad-PDCD5. In protein level, expression of PDCD5 protein in Ad-PDCD5 transfected RAFLS was detected by Western blot. RAFLS transfected with Ad-PDCD5 were cultured in presence or absence of triptolide and RAFLs apoptosis was determined by flow cytometry.

RESULT

Transfection of RAFLS with increasing concentration of Ad-PDCD5 (50-300 MOI) resulted in dose-dependent increase of PDCD5 production. Apoptotic cells percentage of no transfection group, Ad-null group and Ad-PDCD5 group were, respectively, (22.41 +/- 3.87)%, (28.77 +/- 12.97)%, and (48.87 +/- 12.69)%. Alternatively, transfection without triptolide stimuli had no effect. The data showed that gene transfection of PDCD5 alone without triptolide was not sufficient to activate RAFLS 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.

摘要

目的

研究程序性细胞死亡因子5（PDCD5）对雷公藤甲素诱导的类风湿关节炎成纤维样滑膜细胞（RAFLS）凋亡的影响。

方法

用Ad-PDCD5转染类风湿关节炎患者体外培养的滑膜细胞。在蛋白质水平，通过蛋白质免疫印迹法检测Ad-PDCD5转染的RAFLS中PDCD5蛋白的表达。将转染Ad-PDCD5的RAFLS在有或无雷公藤甲素的情况下培养，通过流式细胞术检测RAFLS凋亡情况。

结果

用不同浓度（50 - 300 MOI）的Ad-PDCD5转染RAFLS，导致PDCD5表达呈剂量依赖性增加。未转染组、Ad-空载体组和Ad-PDCD5组的凋亡细胞百分比分别为（22.41±3.87）%、（28.77±12.97）%和（48.87±12.69）%。另外，无雷公藤甲素刺激的转染没有效果。数据表明，单独转染PDCD5基因而无雷公藤甲素不足以激活RAFLS凋亡；PDCD5作为凋亡的增强剂而非诱导剂。