结缔组织生长因子诱导瘢痕成纤维细胞增殖的信号转导机制研究

目的：探讨结缔组织生长因子（connective tissue growth factor，CTGF）诱导增生性瘢痕（hypertrophics car，HS）成纤维细胞增殖的信号转导通路。方法：采用3H一胸腺嘧啶核苷（3H-TdR）掺入法观察不同浓度CTGF促Hs成纤维细胞增殖的效应，以Western Blot法检测CTGF刺激HS成纤维细胞0、5、10、15、30、60min后磷酸化及总细胞外信号调节激酶（extracellular-signal regulated kinase，ERK1／2）的表达，以二者的比值AI衡量信号通路活化程度；应用特异性阻断剂PD98059阻断ERK通路，MTT法检测CTGF诱导细胞增殖的变化。结果：CTGF在一定浓度范围内呈浓度依赖性促HS成纤维细胞增殖，ERK1／2在无CTGF刺激时AI为0．0131±0．00361，CTGF刺激HS成纤维细胞5、10、15、30、60min后的AI值分别为0．0221±0．00329，0．131±0．0361，0．209±0．0201，0．171±0．0379，0．0413±0．00361，在15min达高峰；采用P098059选择性阻断ERK1／2通路后（OD值=0．42±0．046）与CTGF刺激纽比较（OD=0．66±O．035），细胞增殖显著受抑（P〈0．01）。结论：ERK1／2是CTGF诱导Hs戍纤维细胞增殖的主要信号通路。

Connective tissue growth factor stimulates hypertrophic scar derived fibroblasts primarily by ERK/MAPK signal pathway

Objective To explore the proliferative signal pathway activated by connective tissue growth factor （CTGF） in hypertrophic scar （HS） derived fibroblasts. Methods 3H-TdR incorporation technique was used to assess the proliferative effect of CTGF in different concentration. The expression of phosphorylated and total ERK1/2 protein 0,5,10,15,30,60rain after CTGF stimulation was semi- quantitatively analyzed by use of Western-Blotting,and the activation index （AI）was defined to measure the activation of ERK1/2 signal pathway. PD98059 was admitted to specifically block the ERK1/2 pathway, followed by the administration of CTGF, and subsequent cell proliferation was studied by means of MFF. Results CTGF could stimulate fibroblasts proliferation with a dose-dependant manner in certain scale, by activating the ERK1/2 signal pathway building up the AI to the maximal value（0.209±0.0201 ）15rain after CTGF treatment. Inhibition of ERK1/2 by PD98059 suppressed CTGF-mediated HS fibroblasts proliferation significantly, while OD droop from 0.66＋0.035 to 0.42±0.046, P〈0.01. Conclusions CTGF induced a proliferative response in HS fibroblasts, and this action is mainly dependent on the activation of ERK1/2 signal pathway.