摘要
目的探讨结缔组织生长因子（connective tissue growth factor，CTGF）诱导增生性瘢痕（hypertrophic scar，HS）成纤维细胞增殖的信号转导通路。方法采用。H-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印迹法检测CTGF刺激HS成纤维细胞0、5、10、15、30、60min后，磷酸化及非磷酸化细胞外信号调节激酶1／2（extracellular-signal regulated kinase，ERK1／2）的表达，以两者的比值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±0．0036，CTGF刺激HS成纤维细胞5、10、15、30、60min后，AI值分别为0．0221±0．0033，0．1310±0．0361，0．2090±0．0201，0．1710±0．0379，0．0413±0．0036，在15min达高峰；采用PD98059选择性阻断ERK1／2通路后（A=0．420±0．046）与CTGF刺激组比较（A=0．660±0．035），细胞增殖显著受到抑制（P〈0．01）。结论ERK1／2是CTGF诱导HS成纤维细胞增殖的主要活化的信号通路。
Objective To explore the signal mechanism of proliferation stimulating effect of connective tissue growth factor （CTGF） on hypertrophic scar （HS） derived fibroblasts. Methods^3H-TdR incorporation technique was used to determine the proliferative effect of CTGF at different concentration. Western blot was applied to semi-quantitively analyze the expression of phosphorylated and total ERK1/2 protein after 0, 5, 10, 15, 30, and 60 rain of CTGF stimulation, and the relative value of which was defined as AI to measure the activation of ERK1/2 signal pathway. PD98059 was admitted to specifically block the ERK1/2 pathway, and subsequently cell proliferation stimulated by CTGF was studied by MTT. Results CTGF could stimulate fihroblasts proliferation with a dose-dependant manner, and activated the ERK1/2 signal pathway, and AI built up to 0. 209 ±0. 0201, reaching the apex at 15 min after stimulation performed. Inhibition of ERK1/2 activation by PD98059 suppressed CTGF-mediated HS fibroblasts proliferation significantly, while OD significantly dropped. Conclusion CTGF induces a proliferative response in HS fihroblasts, and this action is mainly dependent on the activation of ERK1/2 signal pathway.
出处
《中华医学美学美容杂志》
2009年第3期188-191,共4页
Chinese Journal of Medical Aesthetics and Cosmetology
基金
国家自然科学基金资助项目（编号：30400472）
关键词
结缔组织生长因子
增生性瘢痕
成纤维细胞
细胞外信号调节激酶
信号通路
Connective tissue growth factor
Hypertrophic scar
Fibroblasts
Extracellular-signal regulated protein kinase
Signal pathway
