成纤维细胞生长因子2对凋亡成纤维细胞中S100蛋白表达的影响  

The role of in fibroblast growth factor-2 on expression of S100 protein in apoptosis of fibroblasts after heat stress

作　　者：程飚[1] 刘宏伟[1] 付小兵[2] 盛志勇[2] 张萍[3] 孟晋红[3] 

机构地区：[1]广州军区广州总医院整形科,510010 [2]解放军总医院第一附属医院全军创伤修复重点实验室 [3]第四军医大学全军神经科学研究所

出　　处：《中华烧伤杂志》2006年第2期109-112,共4页Chinese Journal of Burns

基　　金：国家重点基础研究发展计划资助项目(2005CB522603);国家自然科学基金资助项目(30170966;30570575);国家杰出青年科学基金资助项目(39525024);国家博士后科学基金资助项目[中博基(2001)14]

摘　　要：目的观察热损伤后成纤维细胞凋亡模型中,外用成纤维细胞生长因子2(FGF2)对凋亡细胞S100蛋白表达的影响。方法用含体积分数5％小牛血清的DMEM溶液培养已大致融合的成纤维细胞24 h,置入45℃恒温水浴锅中孵育10 min,建立热损伤细胞模型(热损伤组,3瓶),在上述基础上立即加入FGF2(FGF2组,10μg/L,3瓶)。两组细胞继续常规培养30 min后作免疫荧光双标记,利用激光扫描共聚焦显微镜观察细胞半胱氨酸天冬氨酸蛋白酶3(caspase-3)和增殖细胞核抗原(PCNA)以及S100蛋白、磷酸酪氨酸蛋白(P-tyr)、src基因产物蛋白同源的酪氨酸激酶催化功能区-2(SH2-P)表达的变化。结果热损伤组细胞可以同时表达caspase-3蛋白和PCNA两种抗体, 其灰度值分别为140±31、52±8;FGF2组细胞亦见上述两种抗体表达,灰度值分别为75±28、131± 17,PCNA表达量与热损伤组比较,差异有统计学意义(P<0．05)。同时,热损伤组成纤维细胞S100、 P-tyr蛋白、SH2-P蛋白表达阳性,FGF2组细胞S100蛋白和SH2-P的表达明显较弱(P<0．05)、P-tyr 的表达略有减弱。结论离体情况下,热损伤可诱导成纤维细胞凋亡信号的表达;S100蛋白在 FGF2对热损伤导致的凋亡成纤维细胞的增殖和抗凋亡作用中扮演重要角色。Objective To investigate the effects of fibroblast growth factor-2 （FGF2） on S100 protein expression and the involvement of S100 protein in FGF-induced cell proliferation and anti-apoptosis effects in cultured fibroblasts after heat injury in vitro. Methods Human fibreblasts were cultured in Dulbecco＇s modified Eagle＇s medium supplemented with 5% calf serum （DMEM） at 37 ℃ in a humidified atmosphere of 5% CO2 and 95% air for 24 hours, then the cells were injured by heating to 45 ℃ for 10 min. They were divided into two groups. In group one FGF2 （ 10μg/L） was added, and group two served as negative control without the addition of FGF2. The cells were incubated at 37 ℃/5% CO2 for 30 rain. Immunofluorescence double labeling of fibroblasts was carried out and the cells were observed under laser scanning confocal microscope. The intensity of fluorescence label in fibroblasts was measured using NIH Imagine software. Resuits Heat injured fibroblasts were found to be able to express both caspase-3 and proliferating cell nuclear protein（PCNA）, with respective gray scale of 140 ± 31 and 52 ± 8. In group one, after the addition of FGF2, the heat injured fibroblasts were found to express these two proteins too, and with respective gray scale of 75 ± 28 and 131 ± 17. There was a statistically significant difference in values of PCNA between two groups（ P 〈 0. 05）. At the same time, there was positive expression of S100 protein, phosphotyrosine （Ptye） and Src homolog region 2 protein（SH2P） in the heat injured fibroblasts. With the addition of FGF2, the expression of S100 protein and SH2P was weaker（ P 〈 0. 05）, and expression of Ptyr was slightly weaker. Conclusion Heat injury can induce apoptotic signals of fibroblasts in vitro. The role of FGF2 in the regulation of proliferation and anti-apoptosis in heat injured fibroblasts may be, at least in part, attributable to S100 proteins expression.

关 键 词：成纤维细胞 成纤维细胞生长因子2 细胞凋亡 细胞增殖 S100蛋白质类 

分 类 号：R641[医药卫生—外科学]