5-氮杂-2-脱氧胞苷对系统性硬皮病皮损成纤维细胞胶原蛋白及相关抑制因子表达的影响

Effects of 5-aza-2-deoxycytidine on the expression of collagen and its inhibitors in fibroblasts from systemic sclerosis patients

目的:探讨5-氮杂-2-脱氧胞苷对系统性硬皮病皮损成纤维细胞胶原蛋白及相关抑制因子表达的影响。方法:首先,体外培养建立疾病组和对照组皮肤成纤维细胞系。然后,采用实时定量RT-PCR技术,分别测定两组成纤维细胞Dnmt1、Dnmt3a、Dnmt3b、MBD1、MBD2、MeCP2、Kaiso等基因的表达水平。最后,观测5-氮杂-2-脱氧胞苷处理前后Col1α2、Smad7、MMP1等基因的表达水平。结果:与健康对照组比较,系统性硬皮病组皮损成纤维细胞Dnmt1、Col1α2表达增高,Smad7、MMP1表达减少(P<0.05)。经5-氮杂-2-脱氧胞苷处理后,系统性硬皮病组皮损成纤维细胞异常表达的Col1α2降至正常水平,而Smad7、MMP1表达也恢复至正常范围。结论:Dnmt1等表观遗传机制可能参与了系统性硬皮病发病过程。5-氮杂-2-脱氧胞苷可能成为系统性硬皮病治疗的新选择。

Objective：To study effects of 5 -aza -2 -deoxycytidine on the expression of collagen and its inhibitors in fibroblasts from systemic sclerosis patients. Methods：Firstly, we established cultured fibroblast cell lines from skin samples of systemic sclerosis patients and healthy controls. Secondly, the mRNA levels of Dnmtl, Dnmt3a, Dnmt3b, MBD1, MBD2, MeCP2 and Kaiso were determined by Real Time RT- PCR. Finally, effects of 5 -aza- 2 -deoxycytidine on the expression of Col1α2 ,Smad7 ,MMP1 were observed. Results：Compared with the results from controls, transcription levels of Dnmtl and Col1α2 were increased, whereas the mRNA levels of Smad7 and MMP1 were lower in the fibroblasts from systemic sclerosis patients. After the treatment of 5 -aza- 2 -deoxycytidine, the fibroblasts from patients produced less Dnmtl and Col1α2, and transcription of Smad7 and MMP1 was normalized. Conclusion： Epigenetie factors such as Dnmtl may be involved in the pathogenesis of systemic sclerosis, and 5 - aza - 2 - deoxycytidine may become a novel method for the treatment of systemic sclerosis.