CAJ | 학술논문

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,观察时问与剂量的时效关系；半定量RT-PCR、实时荧光定量(Real Time)-PCR、Western blot分别检测各组HSF内MCP-1 mRNA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10,100 nmol/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-1mRNA的表达和蛋白的合成,特别是在10 nmol/L的浓度下表达最为显著(P＜0.01)；在10,1 000nmol/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.
목적 탐토외원성P물질(substance P,SP)대고당조건하인피부성섬유세포(human skin fibroblast,HSF)증식급기단핵세포추화단백-1(monocyte chemoattractant protein-1,MCP-1)합성적영향. 방법 용고당DMEM배양기배양HSF；병용SP재불동농도화불동시상점처리,ELISA법검측세포상청액중MCP-1분비수평,관찰시문여제량적시효관계；반정량RT-PCR、실시형광정량(Real Time)-PCR、Western blot분별검측각조HSF내MCP-1 mRNA적표체화단백적합성；세포증식시제합(cell counting Kit-8,CCK-8)검측불동농도SP작용HSF후적증식정황. 결과 경SP처리후8h가검측도MCP-1분비증강(P＜0.05),24h체봉치(P＜0.01),재10,100 nmol/L적농도하분비최다(P＜0.01)；SP가명현증가고당조건하HSF MCP-1mRNA적표체화단백적합성,특별시재10 nmol/L적농도하표체최위현저(P＜0.01)；재10,1 000nmol/L적SP균명현촉진HSF적증식(P＜0.05). 결론 SP가촉진고당조건하HSF적증식급MCP-1적합성,대촉진당뇨병창면수복가능유일정의의.
Objective To investigate effects of exogenous substance P (SP) on proliferation of humau 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 cxpression 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 ＜ 0.05).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.