CAJ | 학술논문

目的观察血竭素高氯酸盐(Dp)对人皮肤成纤维细胞(Fb)生物学特性的影响,探讨其促进创而愈合的机制.方法分离培养人正常Fb,将Dp以不同浓度(0.063、0.125、0.250、0.625、1.250、2.500 mg/L)分别加入培养液,采用噻唑蓝(MTT)比色法检测Dp在不同浓度以及不同时间点对体外培养的Fb增殖作用的影响.通过流式细胞仪,实时荧光定量聚合酶链式反应( real-time PCR)分别检测最适浓度培养条件下Fb的细胞周期变化以及成纤维细胞生长因子(FGF) mRNA的合成表达.结果 Dp在0.625～1.250 mg/L浓度范围内,其吸光度值(0.237±0.012、0.243±0.017)均高于对照组(0.208±0.011)差异有统计学意义(t值分别为2.11、2.23,P＜0.05),此浓度范围可促进Fb增殖,且呈剂量依赖性,在浓度为1.250 mg/L时(A值0.243±0.017),促增殖作用最为显著(t =2.23,P＜0.01),流式细胞仪结果显示,在浓度为1.250 mg/L时,Dp可明显促进Fb通过G1/S及S/G2期限制点,S期及G2/M期细胞与对照组比较明显增多,G0/G1期细胞与对照组比较明显减少(t值分别为4.32、7.53、3.27,P＜0.05).细胞因子mRNA表达测定中,1.250 mg/L Dp组与对照组比较表达明显上调,差异亦有统计学意义(=1.48,p＜0.05).结论 Dp能显著促进Fb增殖,加快Fb周期进程,同时可促进FGF的mRNA表达,可能与血竭促进创面愈合的机制有关.
목적 관찰혈갈소고록산염(Dp)대인피부성섬유세포(Fb)생물학특성적영향,탐토기촉진창이유합적궤제.방법 분리배양인정상Fb,장Dp이불동농도(0.063、0.125、0.250、0.625、1.250、2.500 mg/L)분별가입배양액,채용새서람(MTT)비색법검측Dp재불동농도이급불동시간점대체외배양적Fb증식작용적영향.통과류식세포의,실시형광정량취합매련식반응( real-time PCR)분별검측최괄농도배양조건하Fb적세포주기변화이급성섬유세포생장인자(FGF) mRNA적합성표체.결과 Dp재0.625～1.250 mg/L농도범위내,기흡광도치(0.237±0.012、0.243±0.017)균고우대조조(0.208±0.011)차이유통계학의의(t치분별위2.11、2.23,P＜0.05),차농도범위가촉진Fb증식,차정제량의뢰성,재농도위1.250 mg/L시(A치0.243±0.017),촉증식작용최위현저(t =2.23,P＜0.01),류식세포의결과현시,재농도위1.250 mg/L시,Dp가명현촉진Fb통과G1/S급S/G2기한제점,S기급G2/M기세포여대조조비교명현증다,G0/G1기세포여대조조비교명현감소(t치분별위4.32、7.53、3.27,P＜0.05).세포인자mRNA표체측정중,1.250 mg/L Dp조여대조조비교표체명현상조,차이역유통계학의의(=1.48,p＜0.05).결론 Dp능현저촉진Fb증식,가쾌Fb주기진정,동시가촉진FGF적mRNA표체,가능여혈갈촉진창면유합적궤제유관.
Objective To observe the effects of dracorhodin perchlorate (Dp) on the biological characteristics of human dermal fibroblasts in vitro,and explore the possible therapeutic mechanism on inducing wound healing.Methods Human normal fibroblasts (Fb) were isolated and cultured,and different concentrations of Dp were added into the culture medium.By using methyl thiazol tetrazolium (MTT) colorimetric assay,the effects of Dp at different concentrations and different time points in vitro on proliferation of Fb were analyzed.The changes in the cell cycle of Fb and cytokine expression of fibroblast growth factor (FGF) mRNA synthesis were detected in optimal concentration by flow cytometry and real-time fluorescence quantitative polymerase chain reaction ( rcal-time PCR).Results Dp in the 0.625-1.250 mg/L concentration range could promote Fb proliferation in a dose-dependent manner,and the most significant effect on the proliferation ( t =2.11,2.23,P ＜ 0.01 ) was obtained at a concentration of 1.250 mg/L.1.250 mg/L Dp could promote Fb to pass through G1/S and S/G2 phase restriction point,the number of cells in S phase and G2/M phase was increased significantly compared with the control group,and number of cells in G0/G1 phase was reduced significantly as compared with the control group (t =4.32,7.53,3.27,P ＜ 0.05 ).As compared with the control group,the cytokine mRNA expression was significantly increased in Fb treated with 1.250 mg/L Dp ( t =1.48,P ＜ 0.05 ).Conclusion Dp can significantly promote the proliferation of Fb,accelerate the process of Fb cycle,and promote the mRNA expression of FGF at the same time,which might be contributed to the mechanisms of Dp promoting wound healing.