实用医学杂志 ›› 2023, Vol. 39 ›› Issue (14): 1733-1739.doi: 10.3969/j.issn.1006⁃5725.2023.14.002

• 基础研究 • 上一篇    下一篇

大黄素通过抑制IGF⁃1介导的Akt/FoxO1信号通路调控人角质形成细胞脂质分泌的机制 

邓方祺1，2 刘思1，2 罗小华1，2 柳宇峰1，2 施歌1，2    

  1. 1 中山大学附属第六医院医学美容整形中心（广州510655）；2 广州市黄埔区中六生物医学创新研究院 （广州510799）
  • 出版日期:2023-07-25 发布日期:2023-07-25
  • 通讯作者: 施歌 E⁃mail：shig9@ mail.sysu.cn
  • 基金资助:
    国家自然科学基金项目（编号：81773345） 

Effects of emodin on proliferation，cell cycle，apoptosis and lipid synthesis in HaCaT keratinocytes 

DENG Fangqi*，LIU Si，LUO Xiaohua，LIU Yufeng，SHI Ge.   

  1. The Sixth Affiliated Hospital，Sun Yat ⁃ sen University， Guangzhou 510655，China 

  • Online:2023-07-25 Published:2023-07-25
  • Contact: SHI Ge E⁃mail：shig9@ mail.sysu.cn

摘要：

目的 探究中药单体大黄素（Emodin）对 HaCaT 人永生化角质形成细胞增殖、周期、凋亡、皮 脂合成与分泌的影响。方法 用 0、25、50、100 μmol/L 大黄素处理 HaCaT，行 CCK⁃8 细胞活力检测、结晶紫 染色观察细胞集落、TUNEL 荧光染色检测细胞集落、流式细胞术检测细胞周期与凋亡、油红 O 染色观察并 定量检测皮脂分泌水平、Western blot 检测蛋白表达；利用胰岛素样生长因子⁃1（IGF⁃1）诱导体外痤疮模型， 检测皮脂分泌情况及 Akt/FoxO1 通路分子蛋白表达水平。结果 大黄素处理组的 HaCaT 细胞融合度及集落形成数量随药物浓度增加而逐渐减少，HaCaT 细胞活力降低，且具有浓度依赖性（P < 0.001），PCNA 表 达水平降低（P < 0.05）；给药组内细胞周期发生 G1/S 阻滞，早期和晚期凋亡率均有所上升，Bax 蛋白表达上 调，Bcl⁃2 蛋白表达下调；细胞内脂滴堆积减少（P < 0.05），成脂因子 PPARγ、SREBP⁃1、LXR 表达水平降低 （P < 0.05）；IGF⁃1 干预组内皮脂分泌增加，皮脂水平显著上调（P < 0.001），Akt 和 FoxO1 蛋白磷酸化程度增 加（P < 0.001）；与IGF⁃1处理组相比，大黄素干预后，细胞皮脂分泌显著减少（P < 0.001），Akt 及FoxO1磷酸 化程度明显降低（P < 0.001）。结论 大黄素能够减少 HaCaT 细胞皮脂合成与分泌，可能与抑制细胞增 殖、阻滞细胞周期、促进细胞凋亡有关，并通过抑制IGF⁃1介导的Akt/FoxO1通路激活阻止痤疮发生。 

关键词: 大黄素, HaCaT 角质形成细胞, 皮脂分泌, 胰岛素样生长因子?1（IGF?1）, 痤疮, Akt/FoxO1信号通路

Abstract:

Objective To investigate the effects of emodin on the proliferation，cell cycle，apoptosis， sebum synthesis and secretion of human HaCaT keratinocytes. Methods HaCaT cells were cultured and treated with emodin（0，25，50，100 μmol/L）. The cellular viability was detected by CCK ⁃ 8 assay. The colony formation was observed by crystal violet staining. Flow cytometry was used to determine cell cycle and apoptosis. Lipid content levels were visualized and analyzed by Oil Red O staining. Western blot was applied for detection of protein expression. Moreover，we used insulin⁃like growth factor⁃1（IGF⁃1）to stimulate lipid synthesis，and then the protein levels Akt/FoxO1 pathway molecules were quantified by western blot analysis. Results Compared to the control group，the cell confluence，numbers of colony formation as well as the viability of HaCaT cells in emodin⁃treated groups were significantly decreased in a dose⁃dependent manner（P < 0.001），while the expression of PCNA was downregulating（P < 0.05）. Flow cytometry analysis revealed that emodin induced G1/S arrest and cell apoptosis in HaCaT cells. The proportion of cell population in early and late apoptosis was markedly increased after emodin treatment，along with upregulation of pro⁃apoptotic protein Bax，downregulation of anti⁃apoptotic protein Bcl⁃2. Our results also showed that emodin reduced the accumulation of intracellular lipid droplets in HaCaT cells（P < 0.05）， and the expression levels of lipid factors（PPARγ，SREBP⁃1 and LXR）were decreased，as well（P < 0.05）. IGF⁃1 enhanced the lipid production（P < 0.001）and the phosphorylation of Akt and FoxO1（P < 0.001）when compared to the untreated group. However，emodin could reverse IGF⁃1⁃induced lipid overproduction and elevated phosphory⁃ lated levels of Akt and FoxO1（P < 0.001）. Conclusion Emodin could reduce sebum secretion in HaCaT cells，possibly due to its inhibitory effects on growth and lipogenesis on HaCaT cells，including suppressing proliferation， arresting cell cycle，inducing apoptosis，and inhibiting IGF⁃1⁃mediated activation of Akt/FoxO1 pathway. 

Key words: Emodin, human HaCaT keratinocytes, sebum synthesis, insulin?like growth factor?1, acne vulgaris

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