中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2014年
7期
494-498
,共5页
相风梅%魏志平%钟连生%杨青%刘彦群
相風梅%魏誌平%鐘連生%楊青%劉彥群
상풍매%위지평%종련생%양청%류언군
阿维A%细胞凋亡%胰岛素样生长因子结合蛋白质类%血管内皮生长因子%HaCaT细胞
阿維A%細胞凋亡%胰島素樣生長因子結閤蛋白質類%血管內皮生長因子%HaCaT細胞
아유A%세포조망%이도소양생장인자결합단백질류%혈관내피생장인자%HaCaT세포
Acitretin%Apoptosis%Insulin-like growth factor binding proteins%Vascular endothelial growth factors%HaCaT cells
目的 探讨阿维A对HaCaT细胞体外凋亡和胰岛素样生长因子结合蛋白7(IGFBP7)及血管内皮生长因子(VEGF)表达的影响.方法 将10-5、10-6、10-7、10-8 mol/L的阿维A分别作用于HaCaT细胞24、48、72 h后,采用CCK8法检测细胞增殖情况;流式细胞仪检测阿维A对HaCaT细胞凋亡率的影响;Western印迹和RT-PCR法检测阿维A对HaCaT细胞IGFBP7、VEGF蛋白及其mRNA表达的影响.结果 10-8 mol/L阿维A处理HaCaT细胞48 h时表现出对细胞增殖的抑制作用,随着时间延长和药物浓度升高,抗增殖作用亦增强,当药物浓度增加到10-5 mol/L时,48 h和72 h的抑制率分别为39.94％±2.27％和49.77％±1.87％.与对照组相比,10-5 mo1/L阿维A作用48 h后,凋亡率由1.803％±0.313％(对照组)上升至7.617％±0.767％(阿维A组)(P＜0.05),IGFBP7的表达由0.436±0.013上升至0.939±0.040 (P＜ 0.05),IGFBP7 mRNA的表达由0.190±0.056上升至0.872±0.079(P＜ 0.05),VEGF的表达由0.798±0.036下降至0.213±0.032(P＜ 0.05),VEGF mRNA的表达由0.933±0.054下降至0.274±0.041 (P＜ 0.05).结论 阿维A可促进HaCaT细胞的体外凋亡,并可在蛋白及mRNA水平上调IGFBP7及下调VEGF的表达.
目的 探討阿維A對HaCaT細胞體外凋亡和胰島素樣生長因子結閤蛋白7(IGFBP7)及血管內皮生長因子(VEGF)錶達的影響.方法 將10-5、10-6、10-7、10-8 mol/L的阿維A分彆作用于HaCaT細胞24、48、72 h後,採用CCK8法檢測細胞增殖情況;流式細胞儀檢測阿維A對HaCaT細胞凋亡率的影響;Western印跡和RT-PCR法檢測阿維A對HaCaT細胞IGFBP7、VEGF蛋白及其mRNA錶達的影響.結果 10-8 mol/L阿維A處理HaCaT細胞48 h時錶現齣對細胞增殖的抑製作用,隨著時間延長和藥物濃度升高,抗增殖作用亦增彊,噹藥物濃度增加到10-5 mol/L時,48 h和72 h的抑製率分彆為39.94％±2.27％和49.77％±1.87％.與對照組相比,10-5 mo1/L阿維A作用48 h後,凋亡率由1.803％±0.313％(對照組)上升至7.617％±0.767％(阿維A組)(P＜0.05),IGFBP7的錶達由0.436±0.013上升至0.939±0.040 (P＜ 0.05),IGFBP7 mRNA的錶達由0.190±0.056上升至0.872±0.079(P＜ 0.05),VEGF的錶達由0.798±0.036下降至0.213±0.032(P＜ 0.05),VEGF mRNA的錶達由0.933±0.054下降至0.274±0.041 (P＜ 0.05).結論 阿維A可促進HaCaT細胞的體外凋亡,併可在蛋白及mRNA水平上調IGFBP7及下調VEGF的錶達.
목적 탐토아유A대HaCaT세포체외조망화이도소양생장인자결합단백7(IGFBP7)급혈관내피생장인자(VEGF)표체적영향.방법 장10-5、10-6、10-7、10-8 mol/L적아유A분별작용우HaCaT세포24、48、72 h후,채용CCK8법검측세포증식정황;류식세포의검측아유A대HaCaT세포조망솔적영향;Western인적화RT-PCR법검측아유A대HaCaT세포IGFBP7、VEGF단백급기mRNA표체적영향.결과 10-8 mol/L아유A처리HaCaT세포48 h시표현출대세포증식적억제작용,수착시간연장화약물농도승고,항증식작용역증강,당약물농도증가도10-5 mol/L시,48 h화72 h적억제솔분별위39.94％±2.27％화49.77％±1.87％.여대조조상비,10-5 mo1/L아유A작용48 h후,조망솔유1.803％±0.313％(대조조)상승지7.617％±0.767％(아유A조)(P＜0.05),IGFBP7적표체유0.436±0.013상승지0.939±0.040 (P＜ 0.05),IGFBP7 mRNA적표체유0.190±0.056상승지0.872±0.079(P＜ 0.05),VEGF적표체유0.798±0.036하강지0.213±0.032(P＜ 0.05),VEGF mRNA적표체유0.933±0.054하강지0.274±0.041 (P＜ 0.05).결론 아유A가촉진HaCaT세포적체외조망,병가재단백급mRNA수평상조IGFBP7급하조VEGF적표체.
Objective To investigate the in vitro effects of acitretin on the apoptosis and expressions of insulin-like growth factor binding protein 7 (IGFBP7) and vascular endothelial growth factor (VEGF) in HaCaT cells.Methods Cultured HaCaT cells were treated with various concentrations (10-5,10-64,10-7,10-8 mol/L) of acitretin for various durations,with those cultured in acitretin-free medium serving as the control group.Then,CCK-8 assay was performed to evaluate the proliferation of cells after 24-,48-and 72-hour treatment,flow cytometry to detect the apoptosis of HaCaT cells,and Western blot and reverse transcription-PCR to quantify the protein and mRNA expressions of IGFBP7 and VEGF in HaCaT cells,respectively,after 48-hour treatment.Statistical analysis was carried out by one-way analysis of variance and Pearson correlation analysis.Results The proliferation of HaCaT cells was inhibited by the treatment with acitretin,and the inhibitory effect increased with the elevation of concentration and prolongation of treatment duration of acitretin.A significant decrease was observed in the proliferative activity of HaCaT cells treated with acitretin of 10-8 mol/L for 48 hours,and when the concentration of acitretin was 10-5 mol/L,the proliferation of HaCaT cells was inhibited by 39.94％ ± 2.27％ and 49.77％ ± 1.87％ at 48 and 72 hours respectively,compared with the control cells.The HaCaT cells treated with acitretin of 10-5 mol/L for 48 hours showed a significant elevation in apoptosis rate (7.617％ ± 0.767％ vs.1.803％ ± 0.313％,P ＜ 0.05),IGFBP7 protein and mRNA expressions (0.939 ± 0.040 vs.0.436 ± 0.013,0.872 ± 0.079 vs.0.190 ± 0.056,both P ＜ 0.05),but a significant reduction in VEGF protein and mRNA expressions (0.213 ± 0.032 vs.0.798 ± 0.036,0.274 ± 0.041 vs.0.933 ± 0.054,both P ＜ 0.05) in comparison to the control cells.Conclusions Acitretin can induce the apoptosis of HaCaT cells,and up-regulate IGFBP7 but down-regulate VEGF expressions in HaCaT cells at protein and mRNA levels.
