瘦素通过激活STAT3信号通路促进HaCaT细胞增殖  

Leptin increases the proliferation of human HaCaT keratinocytes through activation of STAT3 pathway

作　　者：薛柯[1] 刘海燕[2] 简强[1] 张敏[1] 李承新[1] 

机构地区：[1]第四军医大学西京皮肤医院,西安710032 [2]兰州军区兰州总医院皮肤科

出　　处：《中华皮肤科杂志》2013年第12期901-903,共3页Chinese Journal of Dermatology

摘　　要：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印迹分析瘦索激活的下游信号分子活化程度。采用GraphPadPrism5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，50μg，L及100μg／L瘦素作用24及48h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，100μg／L瘦素作用24h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，100μg，L瘦素可使HaCaT细胞STAT3的磷酸化程度明显增高。STAT3抑制剂Diceatannol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。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 ceils treated with leptin of 100 μg/L, Western blot to determine the phosphorylation level of Erkl/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）. Pieeatannol 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 Ixg/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.

关 键 词：T细胞增殖 STAT3 瘦素 Western印迹分析 信号通路 激活 HACAT细胞 人角质形成细胞 

分 类 号：R341[医药卫生—基础医学]