中华皮肤科杂志 ›› 2025, e0250063.doi: 10.35541/cjd.20250063

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痤疮丙酸杆菌通过诱导角质形成细胞发生Gasdermin E介导的焦亡促进痤疮炎症反应

张峰源1    陈思涵2    周子桢1    周蒙2    曾荣2    陈旭2    练霓2    李岷2   

  1. 1北京协和医学院  中国医学科学院  皮肤病医院  皮肤病研究所，南京  210042；2江苏省皮肤医学重点实验室，中国医学科学院  北京协和医学院皮肤病医院，南京  210042
  • 收稿日期:2025-02-10 修回日期:2025-05-20 发布日期:2025-05-30
  • 通讯作者: 李岷 E-mail:limin@pumcderm.cams.cn
  • 基金资助:
    国家自然科学基金（82173432）；达尔肤痤疮研究基金（202335）

Cutibacterium acnes promotes inflammation in acne by inducing Gasdermin E-mediated pyroptosis in keratinocytes

Zhang Fengyuan1, Chen Sihan2, Zhou Zizhen1, Zhou Meng2, Zeng Rong2, Chen Xu2, Lian Ni2, Li Min2   

  1. 1Hospital for Skin Diseases, Institute of Dermatology, Chinese Academy of Medical Sciences ＆ Peking Union Medical College, Nanjing 210042, China; 2Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIs, Hospital for Skin Diseases, Chinese Academy of Medical Sciences ＆ Peking Union Medical College, Nanjing 210042, China
  • Received:2025-02-10 Revised:2025-05-20 Published:2025-05-30
  • Contact: Li Min E-mail:limin@pumcderm.cams.cn
  • Supported by:
    National Natural Science Foundation of China （82173432）; DR.WU Foundation for Acne （202335）

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摘要： 【摘要】 目的 探究痤疮丙酸杆菌（C.acnes）诱导角质形成细胞发生Gasdermin E（GSDME）介导的焦亡在痤疮发病中的作用。方法 用灭活的C.acnes刺激人永生化的角质形成细胞HaCaT 15 min至24 h，采用Western印迹法检测不同时间点HaCaT细胞中GSDME剪切体（GSDME-NT）的表达水平。收集2024年1月2日至12月1日在中国医学科学院皮肤病医院就诊的5例痤疮患者及4例健康对照的皮肤组织样本，另收集痤疮囊肿内容物样本和正常皮肤毛囊内容物样本各3份。分别采用免疫组化法及Western印迹法检测GSDME-NT在表皮中的表达。酶联免疫吸附试验（ELISA）检测痤疮囊肿或正常毛囊内容物中白细胞介素（IL）1β、肿瘤坏死因子（TNF）α的水平。通过转染载有GSDME-shRNA的慢病毒构建GSDME敲低的HaCaT细胞，转染载有无义序列对照（NC）慢病毒的细胞作为对照；ELISA检测GSDME敲低细胞经C.acnes刺激后（C.acnes + GSDME敲低组）IL-1β和TNF-α分泌情况，同时检测PBS + NC组、C.acnes + NC组和PBS + GSDME敲低组中上述炎症因子水平。用视黄醇预处理HaCaT细胞，采用Western印迹法检测视黄醇预处理与未处理的HaCaT细胞经C.acnes刺激后GSDME-NT的表达水平。结果 C.acnes体外刺激HaCaT细胞1 h开始出现GSDME剪切活化，胞内可检测到GSDME-NT。与对照表皮相比，痤疮患者皮损表皮中GSDME-NT阳性的细胞比例（9.34% ± 2.92%比3.05% ± 1.14%，t = -3.47， P = 0.026）及GSDME-NT蛋白表达水平（t = -3.51，P = 0.025）均明显较高。痤疮囊肿内容物中细胞因子IL-1β［1 337.24（1 182.32，2 230.61） pg/ml］、TNF-α［811.31（438.26，817.73）pg/ml］水平显著高于对照正常皮肤毛囊内容物［IL-1β：0.00（0.00，108.21）pg/ml，Z =1.99，P = 0.046；TNF-α：46.67（12.41，53.21）pg/ml，Z = 1.96，P = 0.049］。ELISA检测显示，C.acnes + NC组上清液中IL-1β ［（12.12 ± 3.07） pg/ml］、TNF-α水平［（26.06 ± 1.57） pg/ml］显著高于PBS + NC组［（3.73 ± 2.24） pg/ml、（10.14 ± 0.79） pg/ml；P = 0.003、＜0.001］；与C.acnes + NC组相比，C.acnes + GSDME敲低组上述细胞因子水平［（3.38 ± 0.93） pg/ml、（12.67 ± 2.10） pg/ml］显著降低（P = 0.003、＜0.001）。视黄醇 + C.acnes组细胞中GSDME-NT表达低于C.acnes组（P = 0.029）。结论 C. acnes可能诱导角质形成细胞发生GSDME介导的焦亡，进而促进炎症因子释放，加重痤疮炎症反应。视黄醇或可抑制上述过程。

关键词: 痤疮, 寻常, 痤疮丙酸杆菌, 细胞焦亡, HaCaT细胞, Gasdermin E蛋白, 白细胞介素1β, 肿瘤坏死因子α, 视黄醇

Abstract: 【Abstract】 Objective To investigate the role of Gasdermin E （GSDME）-mediated keratinocyte pyroptosis induced by Cutibacterium acnes （C.acnes） in the pathogenesis of acne. Methods The human immortalized keratinocyte HaCaT cells were stimulated with heat-inactived C.acnes for 15 minutes to 24 hours, and Western blot analysis was performed to determine the expression of cleaved GSDME（GSDME-NT）in HaCaT cells at different time points. Skin tissue samples were collected from 5 acne patients and 4 healthy controls, who visited the Hospital for Skin Diseases, Chinese Academy of Medical Sciences from January 2 to December 1, 2024; additionally, 3 samples of acne cyst contents and 3 samples of normal follicle contents were collected. Immunohistochemical study and Western blot analysis were conducted to determine GSDME-NT expression in the epidermis. Enzyme-linked immunosorbent assay （ELISA） was performed to detect levels of interleukin （IL）-1β and tumor necrosis factor （TNF）-α in acne cyst or normal follicle contents. GSDME-knockdown HaCaT cells were constructed by transfection with lentivirus carrying GSDME-shRNA, and HaCaT cells transfected with lentivirus carrying the nonsense sequence control （NC） served as controls; ELISA was performed to detect the levels of IL-1β and TNF-α in GSDME-knockdown HaCaT cells after C. acnes stimulation （C. acnes + GSDME knockdown group）, as well as in a phosphate-buffered saline （PBS） + NC group, a C. acnes + NC group, and a PBS + GSDME knockdown group. Western blot analysis was conducted to determine the GSDME-NT expression in HaCaT cells pretreated with or without retinol after C. acnes stimulation. Results The cleavage of GSDME in HaCaT cells began at 1 hour after in vitro C. acnes stimulation, with the intracellular detection of GSDME-NT. Compared with the control epidermis, the proportion of GSDME-NT-positive HaCaT cells （9.34% ± 2.92% vs. 3.05% ± 1.14%, t = -3.47, P = 0.026） and GSDME-NT protein expression levels （t = -3.51, P = 0.025） significantly increased in the lesional epidermis of acne patients. The levels of IL-1β （1,337.24 ［1,182.32, 2,230.61］ pg/ml） and TNF-α （811.31 ［438.26, 817.73］ pg/ml） were significantly higher in the acne cyst contents than in the normal follicle contents （IL-1β: 0.00 ［0.00, 108.21］ pg/ml, Z = 1.99, P = 0.046; TNF-α: 46.67 ［12.41, 53.21］ pg/ml, Z = 1.96, P = 0.049）. ELISA showed that the levels of IL-1β and TNF-α were significantly higher in the C. acnes + NC group （12.12 ± 3.07 pg/ml, 26.06 ± 1.57 pg/ml, respectively） than in the PBS + NC group （3.73 ± 2.24 pg/ml, 10.14 ± 0.79 pg/ml, P = 0.003, < 0.001, respectively）; compared with the C. acnes + NC group, the levels of IL-1β and TNF-α significantly decreased in the C. acnes + GSDME knockdown group （3.38 ± 0.93 pg/ml, 12.67 ± 2.10 pg/ml, P = 0.003, < 0.001, respectively）. The GSDME-NT expression was significantly lower in the retinol + C. acnes group than in the C. acnes group （P = 0.029）. Conclusion C. acnes may induce GSDME-mediated pyroptosis in keratinocytes, thereby promoting the release of inflammatory factors and aggravating the inflammatory response in acne. Retinol may inhibit this process.

Key words: Acne vulgaris, Propionibacterium acnes, Pyroptosis, HaCaT cells, GSDME, Interleukin-1beta, Tumor necrosis factor-alpha, Retionl

引用本文

张峰源 陈思涵 周子桢 周蒙 曾荣 陈旭 练霓 李岷. 痤疮丙酸杆菌通过诱导角质形成细胞发生Gasdermin E介导的焦亡促进痤疮炎症反应[J]. 中华皮肤科杂志, 2025,e0250063. doi:10.35541/cjd.20250063

Zhang Fengyuan, Chen Sihan, Zhou Zizhen, Zhou Meng, Zeng Rong, Chen Xu, Lian Ni, Li Min. Cutibacterium acnes promotes inflammation in acne by inducing Gasdermin E-mediated pyroptosis in keratinocytes[J]. Chinese Journal of Dermatology,2025,e0250063. doi:10.35541/cjd.20250063