CAJ | 학술논문

目的探讨瘦素对人角质形成细胞的生物学作用及其分子机制.方法以人永生化角质形成细胞系HaCaT细胞为研究对象,不同浓度人重组瘦素处理细胞,CCK-8法检测瘦素对细胞增殖影响,流式细胞仪检测细胞周期变化,Western印迹分析瘦素激活的下游信号分子活化程度.采用GraphPad Prism 5软件进行统计分析,组间差异采用t检验.结果 CCK-8法检测显示,50 μg/L及100 μg/L瘦素作用24及48 h后可使细胞增殖活性不同程度增加,且瘦素在24h内的促增殖效应呈剂量依赖方式(r=0.9989,P＜0.05).流式细胞仪检测发现,与未经瘦素处理的对照组相比,100 μg/L瘦素作用24 h后S期细胞比例增多,而G0/G1期细胞比例减少;处理组细胞增殖指数为0.603±0.0157,显著高于对照组(0.564±0.0144),差异有统计学意义(P＜0.05).Western印迹发现,100 μg/L瘦素可使HaCaT细胞STAT3的磷酸化程度明显增高.STAT3抑制剂piceatannol能明显抑制瘦素刺激的HaCaT细胞促增殖作用.结论瘦素可能通过激活STAT3信号转导途径促进角质形成细胞增殖.
목적 탐토수소대인각질형성세포적생물학작용급기분자궤제.방법 이인영생화각질형성세포계HaCaT세포위연구대상,불동농도인중조수소처리세포,CCK-8법검측수소대세포증식영향,류식세포의검측세포주기변화,Western인적분석수소격활적하유신호분자활화정도.채용GraphPad Prism 5연건진행통계분석,조간차이채용t검험.결과 CCK-8법검측현시,50 μg/L급100 μg/L수소작용24급48 h후가사세포증식활성불동정도증가,차수소재24h내적촉증식효응정제량의뢰방식(r=0.9989,P＜0.05).류식세포의검측발현,여미경수소처리적대조조상비,100 μg/L수소작용24 h후S기세포비례증다,이G0/G1기세포비례감소;처리조세포증식지수위0.603±0.0157,현저고우대조조(0.564±0.0144),차이유통계학의의(P＜0.05).Western인적발현,100 μg/L수소가사HaCaT세포STAT3적린산화정도명현증고.STAT3억제제piceatannol능명현억제수소자격적HaCaT세포촉증식작용.결론 수소가능통과격활STAT3신호전도도경촉진각질형성세포증식.
Objective To estimate the biological effects of leptin on human HaCaT keratinocytes and explore their molecular mechanisms.Methods Cell counting kit-8 (CCK-8) was used to evaluate the proliferation of cultured HaCaT cells treated with different concentrations of leptin for 24 and 48 hours.Some HaCaT cells were classified into four groups to remain untreated,be treated with leptin (100 μg/L) and piceatannol (a specific inhibitor of STAT3 phosphorylation) alone or in combination for 24 hours,respectively,followed by the evaluation of cell proliferation using CCK-8 kit.Flow cytometry was performed to assess cell cycle of HaCaT cells treated with leptin of 100 μg/L,Western blot to determine the phosphorylation level of Erk1/2 and STAT3 in HaCaT cells treated with leptin of 100 μg/L for different durations.Statistical analysis was done by Student's t-test for unpaired data using GraphPad Prism 5 software.Results The proliferation of HaCaT cells was accelerated to different degrees after treatment with leptin of 50 and 100 μg/L for 24 and 48 hours,and the accelerating effect was in a dose-dependent manner within 24 hours (r =0.9989,P ＜ 0.05).Piceatannol apparently inhibited the promotive effect of leptin on the proliferation of HaCaT cells.There was an obvious elevation in the percentage of cells at S phase ((57.70 ± 5.88)％ vs.(42.50 ± 7.55)％,P ＞ 0.05),but a significant decrease in that at G0/G1 phase ((39.70 ± 1.57)％ vs.(45.20 ± 1.44)％,P ＜ 0.05),with a significant increase in proliferation index (0.603 ±0.0157 vs.0.564 ± 0.0144,P ＜ 0.05) in HaCaT cells treated with leptin of 100 μg/L for 24 hours compared with the untreated controls.Western blot showed that leptin of 100 μg/L markedly enhanced the phosphorylation level of STAT3 in HaCaT cells.Conclusion Leptin may upregulate the proliferation of HaCaT cells through activation of STAT3 pathway.