中华皮肤科杂志 ›› 2013, Vol. 46 ›› Issue (12): 901-903.

• 研究报道 • 上一篇    下一篇

瘦素通过激活STAT3信号通路促进HaCaT细胞增殖

薛柯1,刘海燕2,简强3,张敏3,李承新3   

  1. 1. 第四军医大学西京皮肤医院
    2. 第四军医大学西京医院全军皮肤病研究所；兰州军区兰州总医院皮肤科
    3. 第四军医大学西京医院全军皮肤病研究所
  • 收稿日期:2012-12-26 修回日期:2013-07-23 出版日期:2013-12-15 发布日期:2013-12-01
  • 通讯作者: 李承新 E-mail:li.chengxin@yahoo.cn

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

  • Received:2012-12-26 Revised:2013-07-23 Online:2013-12-15 Published:2013-12-01

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摘要： 【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24 h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。

关键词: 角质形成细胞, 瘦素, STAT3, 细胞增殖

Abstract: XUE Ke*, LIU Hai-yan, JIAN Qiang, ZHANG Min, LI Cheng-xin. *Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi′an 710032, China Corresponding author: LI Cheng-xin, Email: chengxinderm@163.com 【Abstract】 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.

Key words: keratinocytes, Cell proliferation