P物质对高糖条件下成纤维细胞增殖及其单核细胞趋化蛋白-1合成的调节作用  

Regulative effect of substance P on proliferation of human skin fibroblast and expression of its monocyte chemoattractant protein-1 under high glucose condition

作　　者：贾志刚[1] 方勇[1] 姚敏[1] 倪涛[1] 李晓光[1] 石有振[1] 

机构地区：[1]上海交通大学医学院附属第三人民医院烧伤整形科,创伤医学研究所,201900

出　　处：《中华创伤杂志》2012年第12期1125-1129,共5页Chinese Journal of Trauma

基　　金：上海市卫生局基金资助项目（2009102）;上海市卫生局青年基金资助项目（2008Y063）;上海交通大学医学院附属第三人民医院院基金资助项目（sy208-03）

摘　　要：目的探讨外源性P物质（substance P，SP）对高糖条件下人皮肤成纤维细胞（human skin fibroblast, HSF）增殖及其单核细胞趋化蛋白-1（monocyte chemoattractant protein - 1,MCP-1）合成的影响。方法用高糖DMEM培养基培养HSF；并用SP在不同浓度和不同时相点处理，ELISA法检测细胞上清液中MCP-1分泌水平，观察时间与剂量的时效关系；半定量RT—PCR、实时荧光定量（RealTime）-PCR、Westernblot分别检测各组HSF内MCP-1mRNA的表达和蛋白的合成；细胞增殖试剂盒（cell counting Kit-8，CCK-8）检测不同浓度SP作用HSF后的增殖情况。结果经SP处理后8h可检测到MCP-1分泌增强（P〈0．05），24h达峰值（P〈0．01），在10，100nmol／L的浓度下分泌最多（P〈0．01）；SP可明显增加高糖条件下HSF MCP-1 mRNA的表达和蛋白的合成，特别是在10nmol／L的浓度下表达最为显著（P〈0．01）；在10，1000nmol／L的SP均明显促进HSF的增殖（P〈0．05）。结论SP可促进高糖条件下HSF的增殖及MCP-1的合成，对促进糖尿病创面修复可能有一定意义。Objective To investigate effects of exogenous substance P （SP） on proliferation of human skin fibroblast （HSF） and expression of its monocyte chemoattractant protein-1 （ MCP-1 ） under high glucose condition. Methods HSF cultured in high glucose DMEM medium were treated with SP of various concentrations at different time points. Levels of MCP-1 in the supernatants were determined by ELISA and time-effect relationship of SP regulating expression of MCP-1 was analyzed. Expression of MCP-1 mRNA in HSF was detected by semi-quantitative RT-PCR and Real Time-PCR, Conclusion, while expression of MCP-1 by Western blot. Proliferation of HSF treated with SP of different concentrations was detected by cell counting Kit-8 （CCK-8）. Results MCP-1 had a strong expression at 8 hours after SP disposal （P 〈 0.05） and reached the peak at 24 hours （P 〈 0.01 ）. Meanwhile, MCP-1 expressed the most at 10 nmol/L and 100 nmol/L SP （P 〈0.01 ）. SP significantly enhanced the expression of MCP-1 mRNA and its synthesis under high glucose condition, especially at the concentration of 10 nmol/L （ P 〈 0.01 ）. Also, SP induced obvious proliferation of HSF at concentrations of 10 nmol/L and 1 000 nmol/L （P 〈 O. 05）. Conclusion SP promotes proliferation of HSF and expression of MCP-1 in high glucose DMEM medium, which may be of significance in promoting diabetic wound healing.

关 键 词：P物质 成纤维细胞 单核细胞化学吸引蛋白质类 伤口愈合 

分 类 号：R587.1[医药卫生—内分泌] R641[医药卫生—内科学]