中华皮肤科杂志 ›› 2019, Vol. 52 ›› Issue (4): 248-252.doi: 10.3760/cma.j.issn.0412-4030.2019.04.005

• 论著 • 上一篇    下一篇

异维A酸对肽聚糖诱导的SZ95人皮脂腺细胞相关炎症基因表达的影响

曹珂1    侯霄枭1    李昕1    Christos C. Zouboulis2    鞠强1   

  1. 1上海交通大学医学院附属仁济医院皮肤科  200127； 2Departments of Dermatology, Venereology, Allergology and Immunology, Dessau Medical Center, Brandenburg Medical School Theodore Fontane，Dessau  06847, Germany
  • 收稿日期:2018-11-02 修回日期:2019-02-01 发布日期:2019-04-01
  • 通讯作者: 鞠强 E-mail:qiangju@aliyun.com
  • 基金资助:
    国家自然科学基金（81472894）

Effect of isotretinoin on peptidoglycan-induced expression of inflammatory genes in human SZ95 sebocytes

Cao Ke1, Hou Xiaoxiao1, Li Xin1, Christos C. Zouboulis2, Ju Qiang1   

  1. 1Department of Dermatology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200127, China;  2Departments of Dermatology, Venereology, Allergology and Immunology, Dessau Medical Center, Brandenburg Medical School Theodore Fontane, Dessau 06847, Germany
  • Received:2018-11-02 Revised:2019-02-01 Published:2019-04-01
  • Contact: Ju Qiang E-mail:qiangju@aliyun.com
  • Supported by:
    National Natural Science Foundation of China （81472894）

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摘要： 目的 观察异维A酸体外对肽聚糖诱导的SZ95人皮脂腺细胞中与痤疮发病可能相关的炎症基因表达的影响，探讨异维A酸治疗痤疮的分子机制。方法 将培养的SZ95细胞分成3组，对照组不做处理，另外两组用20 mg/L肽聚糖单独（肽聚糖组）或联合10-5 mol/L异维A酸（共刺激组）共同处理。作用3 h后，实时荧光定量PCR检测上述各组SZ95细胞中白细胞介素（IL）-1α、IL-1β、IL-6、IL-8、肿瘤坏死因子α（TNF-α）以及Toll样受体2（TLR2）和其下游基因MyD88 mRNA表达水平；作用24 h后酶联免疫吸附试验检测各组细胞培养上清液中IL-1α、IL-1β、IL-6、IL-8、 TNF-α蛋白表达水平；作用48 h后Western印迹检测TLR2和MyD88蛋白表达水平。采用SPSS 23软件， 3组间指标的比较采用单因素方差分析，并采用Bonferroni校正进行两两比较。结果 对照组、肽聚糖组和共刺激组IL-1α、IL-1β、IL-6、IL-8及TNF-α mRNA和蛋白表达水平差异均有统计学意义（均P < 0.01）；其中，肽聚糖组上述指标均高于对照组及共刺激组，差异均有统计学意义（均P < 0.016 7）。上述3组间MyD88 mRNA表达差异也均有统计学意义（分别为6.707 ± 0.950、10.270 ± 0.477、7.892 ± 0.900，F = 10.17，P < 0.01），肽聚糖组高于对照组及共刺激组，差异均有统计学意义（t值分别为4.740、3.298，均P < 0.016 7）。肽聚糖组TLR2 mRNA和蛋白表达水平均高于对照组，但与共刺激组比较差异无统计学意义。结论 异维A酸抑制肽聚糖诱导的SZ95人皮脂腺细胞中与痤疮发病可能相关的炎症因子的表达，其机制可能是通过抑制天然免疫应答中MyD88的表达而非TLR2本身，这可能是异维A酸治疗痤疮的机制之一。

关键词: 寻常痤疮； 异维A酸； 药理作用分子作用机制； 肽聚糖； 细胞因子类； Toll样受体2； 免疫, 先天； 人皮脂腺细胞

Abstract: 【Abstract】 Objective To evaluate the effect of isotretinoin on expression of acne-associated inflammatory genes induced by peptidoglycan in human SZ95 sebocytes, and to explore the molecular mechanism underlying the treatment of acne with isotretinoin. Methods Cultured SZ95 sebocytes were divided into 3 groups: control group receiving no treatment, peptidoglycan group treated with 20 mg/L peptidoglycan alone, and costimulation group treated with 20 mg/L peptidoglycan combined with 10-5 mol/L isotretinoin. After 3-hour treatment, real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of pro-inflammatory cytokines including interleukin （IL）-1α, IL-1β, IL-6, IL-8, tumor necrosis factor （TNF）-α, Toll-like receptor 2 （TLR2） and MyD88 （a downstream gene of TLR2） in SZ95 sebocytes in the above groups. After 24-hour treatment, enzyme-linked immunosorbent assay （ELISA） was performed to detect levels of IL-1α, IL-1β, IL-6, IL-8 and TNF-α in the cell culture supernatant in the above groups. After 48-hour treatment, Western blot analysis was conducted to determine the protein expression of TLR2 and MyD88. Statistical analysis was carried out with SPSS 23 software by one-way analysis of variance （ANOVA） for the comparison among the 3 groups, and by Bonferroni method for multiple comparisons. Results The mRNA and protein expression of pro-inflammatory cytokines including IL-1α, IL-1β, IL-6, IL-8 and TNF-α all significantly differed among the 3 groups （all P < 0.01）, and was significantly higher in the peptidoglycan group than in the control group and costimulation group （both P < 0.016 7）. The mRNA expression of MyD88 also significantly differed among the control group, peptidoglycan group and costimulation group （6.707 ± 0.950, 10.270 ± 0.477, 7.892 ± 0.900 respectively, F = 10.17, P < 0.01）, and was significantly higher in the peptidoglycan group than in the control group and costimulation group （t = 4.740, 3.298 respectively, both P < 0.016 7）. The mRNA and protein expression of TLR2 were markedly higher in the peptidoglycan group than in the control group, but did not differ between the peptidoglycan group and the costimulation group. Conclusion Isotretinoin can inhibit peptidoglycan-induced expression of inflammatory factors possibly associated with the occurrence of acne in human SZ95 sebocytes, likely by inhibiting the expression of MyD88, but not TLR2, in the innate immune response, which may be one of the mechanisms underlying the treatment of acne with isotretinoin.

Key words: Acne vulgaris, Isotretinoin, Molecular mechanisms of pharmacological action, Peptidoglycan, Cytokines, Toll-like receptor 2;Immunity, innate, Human sebocytes

引用本文

曹珂 侯霄枭 李昕 Christos C. Zouboulis 鞠强. 异维A酸对肽聚糖诱导的SZ95人皮脂腺细胞相关炎症基因表达的影响[J]. 中华皮肤科杂志, 2019,52(4):248-252. doi:10.3760/cma.j.issn.0412-4030.2019.04.005

Cao Ke, Hou Xiaoxiao, Li Xin, Christos C. Zouboulis, Ju Qiang. Effect of isotretinoin on peptidoglycan-induced expression of inflammatory genes in human SZ95 sebocytes[J]. Chinese Journal of Dermatology, 2019, 52(4): 248-252.doi:10.3760/cma.j.issn.0412-4030.2019.04.005