中国麻风皮肤病杂志 ›› 2021, Vol. 37 ›› Issue (5): 265-270.doi: 10.12144/zgmfskin202105265

• 论著 • 上一篇    下一篇

N-乙酰半胱氨酸对PM2.5引起HaCaT细胞损伤的保护作用

李锦濯,林志鹏,曾倩雯,孙仁山   

  1. 陆军军医大学大坪医院皮肤科，重庆，400042
  • 出版日期:2021-05-15 发布日期:2021-04-29
  • 通讯作者: 孙仁山，E-mail: pharsunr@126.com

Protective effect of N-acetylcysteine on damage of HaCaT cells induced by PM2.5

LI Jinzhuo, LIN Zhipeng, ZENG Qianwen, SUN Renshan   

  1. Department of Dermatology, Daping Hospital, Army Medical University, Chongqing 400042, China
  • Online:2021-05-15 Published:2021-04-29
  • Contact: SUN Renshan, E-mail: pharsunr@126.com

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摘要： 目的：研究N-乙酰半胱氨酸（N-Acetyl-L-cysteine,NAC）对PM2.5诱导的HaCaT细胞损伤的影响。方法：HaCaT细胞分为NAC干预组、不同浓度PM2.5组(25、50、100、200、400 μg/mL)及空白对照组。 DCFH-DA荧光探针法检测HaCaT细胞内活性氧（ROS）的水平，RT-PCR和ELISA法分别检测IL-1β、IL-33、TSLP mRNA表达及蛋白水平。CCK-8法和Western blot法分别检测细胞存活率和NF-κB信号通路相关蛋白表达水平。结果：不同浓度PM2.5组HaCaT细胞ROS水平均显著高于对照组(P<0.05)；200 μg/mL和400 μg/mL PM2.5处理组细胞中IL-1β、TSLP、IL-33 mRNA表达水平显著高于对照组(P<0.05)。与PM2.5组比较，NAC干预组HaCaT细胞存活率增高，ROS水平降低，NF-κB p-p65蛋白表达降低，IL-1β、TSLP、IL-33分泌水平降低，差异有统计学意义(P＜0.05)。结论：NAC对PM2.5诱导的HaCaT细胞损伤具有保护作用，其机制可能与其降低细胞内氧化应激水平有关。

关键词: PM2.5, HaCaT细胞, 细胞内活性氧, N-乙酰半胱氨酸

Abstract: Objective: To investigate the effect of N-acetylcysteine (NAC) on damage of human immortalized keratinocytes(HaCaT cells) induced by PM2.5. Methods: The HaCaT cells were randomly divided into the NAC group, PM2.5 groups with different concentrations (25, 50, 100, 200, 400 μg/mL) and blank control group. The levels of reactive oxygen species (ROS) in HaCaT cells were detected by DCFH-DA fluorescent probe, and the mRNA expression and protein levels of IL-1β, IL-33 and TSLP were detected by RT-PCR and ELISA respectively. CCK-8 and Western blot were used to detect the cell survival rate and the expression of proteins related to NF-κB signaling pathway. Results: The ROS levels of HaCaT cells in PM2.5 groups were significantly higher than those in the control group (P<0.05). The mRNA levels of IL-1β, TSLP and IL-33 in 200 μg/mL and 400 μg/mL PM2.5 groups were significantly higher than those in the control group (P<0.05). Compared with PM2.5 groups, the survival rate of HaCaT cells in the NAC group was higher, ROS, NF-κB p-p65 protein and IL-1β, TSLP and IL-33 level were lower, with significant differences (Ps<0.05). Conclusion: NAC can antagonize the HaCaT cell damage induced by PM2.5, which may be related to the reduction of intracellular oxidative stress.

Key words: PM2.5, HaCaT cells, ROS, N-acetylcysteine