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、400μg/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);200μg/mL和400μg/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。
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(25,50,100,200,400μg/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 200μg/mL and 400μg/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(P s<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.
作者
李锦濯
林志鹏
曾倩雯
孙仁山
LI Jinzhuo;LIN Zhipeng;ZENG Qianwen;SUN Renshan(Department of Dermatology,Daping Hospital,Army Medical University,Chongqing 400042,China)
出处
《中国麻风皮肤病杂志》
2021年第5期265-270,共6页
China Journal of Leprosy and Skin Diseases
基金
国家自然科学基金面上项目(编号:81571569)。
