Protective role of DDIT4-mediated autophagy in dermal papilla cells after oxidative stress injury
10.16016/j.2097-0927.202406063
- VernacularTitle:DDIT4介导的自噬在毛乳头细胞氧化应激损伤中的保护作用
- Author:
Yuanyuan GAO
1
;
Hengguang ZHAO
;
Guihong YANG
;
Xia LEI
Author Information
1. 400042 重庆,陆军特色医学中心皮肤科
- Keywords:
DNA damage induced transcription factor 4;
autophagy;
oxidative stress;
dermal papilla cells
- From:
Journal of Army Medical University
2024;46(21):2397-2406
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the protective role of DNA damage induced transcription factor 4(DDIT4)in oxidative stress injury in dermal papilla cells and its underlying mechanisms.Methods Dermal papilla cells were exposed to UVA and H2O2 to establish cellular model of oxidative stress.CCK-8 assay was used to detect cell viability under different treatment conditions,and the production of intracellular reactive oxygen species(ROS)was detected using 2',7'-dichlorofluorescein diacetate(DCFH-DA).Autophagic vesicles were observed with electron microscopy.Western blotting was employed to measure the expression of DDIT4 and autophagy-related molecules,including microtubule-associated protein 1 light chain 3(LC3),ubiquitin-binding protein(P62),mammalian target of rapamycin(mTOR),and p-mTOR.Results UVA and H2O2 resulted in more production of ROS(P＜0.05)and decreased viability of dermal papilla cells(P＜0.05).DDIT4 expression was increased in dermal papilla cells under oxidative stress(P＜0.05),and the antioxidant N-acetyl-L-cysteine(NAC)could effectively inhibit this effect(P＜0.05).After treatment with UVA or H2O2,cell autophagy was enhanced in dermal papilla cells,characterized by an increase in the number of autophagosomes and an increased LC3 Ⅱ/Ⅰ ratio(P＜0.05),a decrease in P62 expression(P＜0.05),and 3-methyladenine(3-MA)blocking autophagy led to further reduced cell viability(P＜0.05)and increased intracellular ROS production(P＜0.05).Conversely,rapamycin(RAPA)increased autophagy level and improved the viability of dermal papilla cells under oxidative conditions(P＜0.05),and reduced the generation of intracellular ROS(P＜0.05).Additionally,down-regulation of DDIT4 weakened autophagy in dermal papilla cells under oxidative stress,reduced LC3 Ⅱ/Ⅰ(P＜0.05),increased p-mTOR/mTOR and P62(P＜0.05),inhibited cell viability(P＜0.05),and enhanced intracellular ROS production(P＜0.05).Conclusion DDIT4 may alleviate oxidative stress injury in dermal papilla cells through autophagy.
