上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (7): 905-.doi: 10.3969/j.issn.1674-8115.2011.07.008

• 论著（基础研究） • 上一篇    下一篇

机械损伤对人皮肤角质形成细胞氧化应激和前列腺素E2分泌的影响

傅秀军, 石有振, 方 勇, 姚 敏, 王 莹   

  1. 上海交通大学 医学院附属第三人民医院烧伤整形科创伤医学研究所, 上海 201900
  • 出版日期:2011-07-28 发布日期:2011-07-27
  • 通讯作者: 王 莹, 电子信箱: wy04wy@hotmail.com。
  • 作者简介:傅秀军（1985—）, 男, 硕士生；电子信箱: fuxj84@yahoo.com.cn。
  • 基金资助:

    上海市科委基金（10ZR1418400）

Effects of mechanical damage on oxidative stress and prostaglandin E2 secretion in human skin keratinocytes

FU Xiu-jun, SHI You-zhen, FANG Yong, YAO Min, WANG Ying   

  1. Department of Burns and Plastic Surgery, the Third People's Hospital, Institute of Traumatic Medicine, Shanghai Jiaotong University School of Medicine, Shanghai 201900, China
  • Online:2011-07-28 Published:2011-07-27
  • Supported by:

    Shanghai Science and Technology Committee Foundation, 10ZR1418400

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摘要：

目的 探讨机械损伤对人皮肤角质形成细胞前列腺素E2（PGE2）分泌的影响及其可能的机制。方法 将体外培养的人皮肤角质形成细胞（HaCaT）分为损伤组、NADPH氧化酶(NOX)抑制剂氯化二亚苯基碘鎓（DPI）预处理组和抗氧化剂N-乙酰半胱氨酸(NAC)预处理组，三组细胞经划痕法建立体外机械损伤细胞模型，以未建模的HaCaT细胞作为对照组。于建模后不同时点收集损伤组、DPI预处理组和对照组的细胞及各组细胞的培养上清液，分别采用荧光探针技术和化学发光法测定细胞内活性氧（ROS）的生成和NOX活性，以酶联免疫吸附试验检测各组细胞培养上清液中前列腺素E2（PGE2）的质量浓度。结果 建模后各时点，损伤组和DPI预处理组细胞内ROS生成和NOX活性均显著高于对照组（P<0.05或P<0.01）；DPI预处理组建模后各时点细胞内ROS生成和NOX活性均显著低于损伤组（P<0.05或P<0.01）。各组细胞培养上清液中PGE2质量浓度的检测结果显示：建模后各时点，损伤组、DPI预处理组和NAC预处理组均显著高于对照组（P<0.01），DPI预处理组和NAC预处理组均显著低于损伤组（P<0.01）。结论 机械损伤后人皮肤角质形成细胞的PGE2分泌量增加，其机制可能与细胞内ROS生成增加和NOX活性升高所介导的氧化应激有关。

关键词: 人皮肤角质形成细胞, 机械损伤, 氧化应激, 前列腺素E2

Abstract:

Objective To investigate the effects of mechanical damage on prostaglandin E2 (PGE2) secretion in human skin keratinocytes, and explore its possible mechanism. Methods Human skin keratinocytes (HaCaT) cultured in vitro were divided into injury group, NADPH oxidase (NOX) inhibitor diphenyleneiodonium chloride (DPI) pretreatment group and antioxidant N-acetylcysteine (NAC) pretreatment group. Mechanical damage cell models were established by scratch method, and HaCaT cells without model establishment were served as control group. Cells and culture supernatant were collected from injury group, DPI pretreatment group and control group at different time points after model establishment, fluorescent probe and chemiluminescence method were employed to detect the intracellular reactive oxygen species (ROS) generation and NOX activity respectively, and the mass concentration of prostaglandin E2 (PGE2) in culture supernatant in each group was determined by enzyme-linked immunosorbent assay. Results At each time point after model establishment, the intracellular ROS generation and NOX activity in injury group and DPI pretreatment group were significantly higher than those in control group (P<0.05 or P<0.01), and the intracellular ROS generation and NOX activity in DPI pretreatment group were significantly lower than those in injury group (P<0.05 or P<0.01). The mass concentrations of PGE2 in culture supernatant in injury group, DPI pretreatment group and NAC pretreatment group were significantly higher than that in control group （P<0.01）, while the mass concentrations of PGE2 in culture supernatant in DPI pretreatment group and NAC pretreatment group were significantly lower than that in injury group at each time point after model establishment (P<0.01). Conclusion PGE2 secretion in human skin keratinocytes may increase after mechanical damage, which may be associated with oxidative stress mediated by increased intracellular ROS generation and NOX activity.

Key words: human skin keratinocyte, mechanical damage, oxidative stress, prostaglandin E2