中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2010年
2期
101-104
,共4页
康健%陈文琦%夏济平%李燕华%杨波%纪超%宋秀祖%相文忠%万寅生%毕志刚
康健%陳文琦%夏濟平%李燕華%楊波%紀超%宋秀祖%相文忠%萬寅生%畢誌剛
강건%진문기%하제평%리연화%양파%기초%송수조%상문충%만인생%필지강
紫外线%细胞衰老%细胞外基质%丝裂原激活蛋白激酶激酶类
紫外線%細胞衰老%細胞外基質%絲裂原激活蛋白激酶激酶類
자외선%세포쇠로%세포외기질%사렬원격활단백격매격매류
Ultraviolet rays%Cell aging%Extracellular matrix%Mitogen-activated protein kinase kinase
目的 探讨UVB诱导的衰老(UVB-SIPS)成纤维细胞分泌的细胞外基质(ECM)对HaCaT细胞增殖的影响以及细胞外信号调节激酶(ERK)信号途径的作用.方法 采用辐射诱导人皮肤成纤维细胞衰老,分别制备由未衰老和衰老成纤维细胞分泌的ECM包被的培养皿(分别定为PRE-ECM组、SCIP-ECM组).以空白培养皿(NON-ECM组)为参照,在HaCaT细胞接种于培养皿后,采用MTT法和流式细胞法检测其细胞增殖、细胞周期等指标的变化;免疫印迹法分析ERK活化水平.干预性研究ERK信号通路对上述功能的影响.结果 三组中,SCIP-ECM组可诱导最强烈的ERK1/2的活化.采用U0126抑制ERK信号活化可完全抑制衰老成纤维细胞来源的ECM的促细胞增殖效应.阻断HaCaT细胞的ERK活化,NON-ECM组、PRE-ECM组和SCIP-ECM组S+G2/M期细胞的比例明显下降,分别由处理前37.40%、41.34%和43.31%减少至29.41%、36.48%到39.96%.结论 UVB-SCIP成纤维细胞分泌的ECM通过刺激ERK磷酸化促进HaCaT细胞增殖.
目的 探討UVB誘導的衰老(UVB-SIPS)成纖維細胞分泌的細胞外基質(ECM)對HaCaT細胞增殖的影響以及細胞外信號調節激酶(ERK)信號途徑的作用.方法 採用輻射誘導人皮膚成纖維細胞衰老,分彆製備由未衰老和衰老成纖維細胞分泌的ECM包被的培養皿(分彆定為PRE-ECM組、SCIP-ECM組).以空白培養皿(NON-ECM組)為參照,在HaCaT細胞接種于培養皿後,採用MTT法和流式細胞法檢測其細胞增殖、細胞週期等指標的變化;免疫印跡法分析ERK活化水平.榦預性研究ERK信號通路對上述功能的影響.結果 三組中,SCIP-ECM組可誘導最彊烈的ERK1/2的活化.採用U0126抑製ERK信號活化可完全抑製衰老成纖維細胞來源的ECM的促細胞增殖效應.阻斷HaCaT細胞的ERK活化,NON-ECM組、PRE-ECM組和SCIP-ECM組S+G2/M期細胞的比例明顯下降,分彆由處理前37.40%、41.34%和43.31%減少至29.41%、36.48%到39.96%.結論 UVB-SCIP成纖維細胞分泌的ECM通過刺激ERK燐痠化促進HaCaT細胞增殖.
목적 탐토UVB유도적쇠로(UVB-SIPS)성섬유세포분비적세포외기질(ECM)대HaCaT세포증식적영향이급세포외신호조절격매(ERK)신호도경적작용.방법 채용복사유도인피부성섬유세포쇠로,분별제비유미쇠로화쇠로성섬유세포분비적ECM포피적배양명(분별정위PRE-ECM조、SCIP-ECM조).이공백배양명(NON-ECM조)위삼조,재HaCaT세포접충우배양명후,채용MTT법화류식세포법검측기세포증식、세포주기등지표적변화;면역인적법분석ERK활화수평.간예성연구ERK신호통로대상술공능적영향.결과 삼조중,SCIP-ECM조가유도최강렬적ERK1/2적활화.채용U0126억제ERK신호활화가완전억제쇠로성섬유세포래원적ECM적촉세포증식효응.조단HaCaT세포적ERK활화,NON-ECM조、PRE-ECM조화SCIP-ECM조S+G2/M기세포적비례명현하강,분별유처리전37.40%、41.34%화43.31%감소지29.41%、36.48%도39.96%.결론 UVB-SCIP성섬유세포분비적ECM통과자격ERK린산화촉진HaCaT세포증식.
Objective To explore the influences of extracellular matrices (ECM) secreted by ultraviolet B (UVB)-induced senescent fibroblasts on the proliferation of and extracellular signal-regulated kinase (ERK) signaling in HaCaT cells. Methods Fibroblasts were irradiated with UVB of 15 mJ/cm2 once daily for 5 days to induce premature senescence, which was identified by SA-β-gal staining 72 hours after the last irradiation.HaCaT cells were divided into 3 groups and inoculated into plates coated with extracellular matrices secreted by non-senescent (PRE-ECM) or senescent fibroblasts (SIPS-ECM) or into uncoated plates (NON-ECM), fol-lowed by additional culture. U0126, an inhibitor of ERK1/2, was used to treat the HaCaT cells 1 hour before inoculation. Then, MTT assay was carried out to detect the proliferation of HaCaT cells after a 3-day culture,Western blot to assess the phosphorylation of ERK at 0.5, 1, 2 and 4 hours after the inoculation, flow cytometry to analyse cell cycle and apoptosis after 24 hours of culture. Results The most rapid and intense phosphory-lation of ERK was observed in SIPS-ECM group. Inhibiting the activation of ERK pathway with U0126 could completely suppress the promoting effect of ECM from senescent fibroblasts on the proliferation of HaCaT cells.After the blocking of ERK activation, the proportion of HaCaT cells in S and G2/M phase decreased from 37.40%, 41.34% and 43.31% to 29.41%, 36.48% and 39.96%, respectively, in NON-ECM, PRE-ECM and SCIP-ECM group. Conclusion The ECM produced by UVB-induced senescent fibroblasts promote the prolifera-tion of HaCaT cells via inducing the phosphorylation of ERK.
