中国麻风皮肤病杂志 ›› 2019, Vol. 35 ›› Issue (1): 1-5.doi: 10.12144/zgmfskin201901001

• 论著 •    下一篇

α-硫辛酸对UVB诱导的角质形成细胞氧化应激的影响

黄丹1  鞠梅1  陈崑1  吴敏智2  顾恒1   

  1. 1中国医学科学院北京协和医学院皮肤病研究所理疗科、江苏省皮肤病与性病分子生物学重点实验室，南京，210042
    2苏州市第五人民医院,苏州，215007
  • 出版日期:2019-01-15 发布日期:2019-01-21
  • 通讯作者: 陈崑，E-mail: kunchen181@aliyun.com 顾恒，E-mail: doctor_guheng@hotmail.com

Effect of α-lipoic acid on oxidative stress of keratinocytes induced by UVB

HUANG Dan1, JU Mei1, CHEN Kun1, WU Minzhi2, GU Heng1   

  1. 1. Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIs；Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing 210042, China;  2.The Fifth People's Hospital of  Suzhou, 215007, China
  • Online:2019-01-15 Published:2019-01-21
  • Contact: CHEN Kun, E-mail: kunchen181@aliyun.com GU Heng, E-mail: doctor_guheng@hotmail.com

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摘要： 目的：明确抗氧化剂α-硫辛酸(α-lipoic acid, LA)对UVB辐射后人原代角质形成细胞氧化应激的影响。方法：建立UVB辐射和外源性H2O2诱导的氧化应激模型，倒置显微镜下观察UVB和H2O2刺激后细胞形态和显微结构的变化，2'7'-二氯荧光乙酰乙酸钠(2'7'-dichlorofluorescein etheric acid,DCFH-DA)荧光探针对细胞内活性氧片段(Reactive oxygen species,ROS)进行染色，倒置荧光显微镜下观察荧光强度的变化，并用流式细胞术对绿色荧光进行定量。运用化学法检测各组细胞内脂质过氧化物(Malonaldehyde, MDA)和总抗氧化能力(Total antioxidant capacity, TAOC)。结果：UVB照射后4h细胞的荧光强度为35311±3072.3，MDA为(11.61±0.68)nmol/mg·prot，较阴性对照组明显升高［（24465±2085.3，(8.25±1.21)nmol/mg·prot］,差异均具有统计学意义（P＜0.05）。UVB照射加LA干预4 h后荧光强度下降至26926±3502.7，(9.14±1.12)nmol/mg·prot。TAOC在各组间无统计学差异。结论：LA能够减轻UVB照射后相关的氧化应激损伤。

关键词: 角质形成细胞, 中波紫外线, α-硫辛酸, 氧化应激

Abstract: Objective: To determine the effect of α-lipoic acid on human primary keratinocyte oxidative stress induced by UVB radiation. Methods: The oxidative stress model was established by H2O2 and UVB. The cell morphology and fluorescent intensity were observed under inverted microscope or fluorescence microscope. The intracellular ROS was detcted with fluorescent nuclear staining of 2'7'-acetyl chloride fluorescence sodium acetate (DCFH-DA). Green fluorescent was quantified by flow cytometry. The total antioxidant capacity of total antioxidant capacity (TAOC) and malonaldehyde (MDA) were measured by chemical method. Results: The fluorescence intensity and MDA in cells irradiated by UVB for 4hs were 35311±3072.3 and 11.61±0.68 nmol/mg·prot , which were higher than that in negative control group (24465±2085.3 and 8.25±1.21), with significant differences (Ps<0.05). After incubated 4hs with antioxidant LA, the fluorescence intensity and MDA were reduced to 26926±3502.7 and 9.14±1.12 vnmol/mg·prot. There was no significant difference in the level of TAOC between different groups. Conclusion: LA can alleviate the oxidative stress of the keratinocyte induced by UVB irradiation. 

Key words: keratinocyte, ultraviolet B, α-lipoic acid, oxidative stress