摘要
目的:比较结缔组织生长因子(CTGF)在瘢痕疙瘩成纤维细胞(KF)和正常真皮成纤维细胞(NF)中的表达以及对血清刺激的反应;研究介导CTGF在KF中表达的信号途径。方法:在血清刺激前30min给予小分子人工合成的信号通路抑制剂预处理,分别为:PI3K抑制剂wortmannin(100nM);ERK抑制剂PD98059(50mM);p38MAPK抑制剂SB203580(10mM);JNK抑制剂SP600125(25mM);以及TGF-βI型受体抑制剂SB431542(0.1μM、0.5μM、1μM和10μM),空白对照组仅加入溶剂DMSO预处理。使用定量实时反转录聚合酶链反应比较CTGF在瘢痕疙瘩成纤维细胞(KF)和正常皮肤来源的成纤维细胞(NF)中的表达。结果:KF血清刺激后CTGF的表达迅速升高,而在NF血清刺激后仅少量上升。JNK抑制剂SP600125和TGF-βI型受体抑制剂SB431542对CTGF表达的升高具有显著的抑制作用(P<0.05)。结论:KF中的CTGF血清反应需要JNK和TGF-β信号途径的协同作用。
Objective To compare Connective Tissue Growth Factor （CTGF） expression between keloid derived fibroblasts （KFs） and normal dermal fibroblast （NFs） upon serum stimulation, and study the signal transduction pathways. Methods Small molecule synthesized inhibitors of various signal pathways were added to culture medium 30 min before serum stimulation to block specific signal transduction pathways, including PI3K inhibitor wortmannin （100 nM）. ERK inhibitor PD98059 （50 mM）. p38 MAPK inhibitor SB203580 （10 mM）.JNK inhibitor SP600125 （25 mM）,and TGF--β type Ⅰ receptor inhibitor SB431542 （0.1μM, 0.5μM, 1μM and 10μM）. RNA was collected 0, 1, 6, 12 and 24 hours after serum stimulation and the expression of CTGF in KFs and NFs was analyzed with quantitative real-time PCR. Results The expression of CTGF was elevated significantly in KFs versus NFs. JNK inhibitor SP600125 and TGF-β type Ⅰ receptor inhibitor SB431542 decreased the CTGF up-regulation in KFs after serum stimulation significantly （P〈0.05）. Conclusion Both JNK and TGF-19/SMAD signaling pathway took part in the over-expression of CTGF in KFs.
出处
《中国美容医学》
CAS
2009年第4期506-509,共4页
Chinese Journal of Aesthetic Medicine
基金
国家自然科学基金项目(编号:30371469和30801190)
陕西省自然科学基金项目(编号:2006C253)
