饥饿环境中活性氧激活人牙周膜细胞自噬的研究

doi:10.13701/j.cnki.kqyxyj.2022.09.017

口腔医学研究 ›› 2022, Vol. 38 ›› Issue (9): 877-881.DOI: 10.13701/j.cnki.kqyxyj.2022.09.017

饥饿环境中活性氧激活人牙周膜细胞自噬的研究

何海燕¹, 余京泓², 伍宇婕¹, 张闵¹, 徐晓梅^1*

1. 西南医科大学附属口腔医院正畸科四川泸州 646000;
2. 重庆大学附属三峡医院口腔正畸科重庆 404100

收稿日期:2022-01-20 发布日期:2022-09-26
通讯作者: *徐晓梅,E-mail: xuxiaomei@hotmail.com
作者简介:何海燕(1994~ ),女,四川人,硕士,医师,研究方向:口腔正畸学。
基金资助:
四川省科技厅应用基础研究项目(编号:21YYJC3483)泸州市-西南医科大学联合项目(编号:2020LZXNYDZ06)

A Study on Activation of Autophagy in Human Periodontal Ligament Cells by Reactive Oxygen Species in Starvation Environment.

HE Haiyan¹, YU Jinghong², WU Yujie¹, Zhang Min¹, XU Xiaomei^1*

1. Department of Orthodontics, Hospital of Stomatology Southwest Medical University, Luzhou 626000, China;
2. Department of Orthodontics, Chongqing University Three Gorges Hospital, Chongqing 404100, China.

Received:2022-01-20 Published:2022-09-26

摘要/Abstract

摘要： 目的: 研究饥饿环境下人牙周膜细胞(hPDLCs)的自噬水平变化,探讨活性氧(ROS)在自噬过程中的作用。方法: 分离培养hPDLCs,EBSS模拟营养缺乏环境,透射电镜观察自噬小体的产生评估hPDLCs中的自噬水平;抗氧化剂N-乙酰半胱氨酸(NAC)预处理抑制ROS的生成,DCFH-DA染色检测ROS水平,RT-qPCR、Western blot检测自噬相关蛋白LC3-Ⅱ、Beclin-1、p62表达情况,探究饥饿环境下自噬激活的调控机制。结果: 经EBSS饥饿培养,hPDLCs自噬激活。NAC通过抑制ROS生成,部分逆转了饥饿的hPDLCs中的自噬水平。结论: ROS作为信号分子介导自噬,饥饿通过诱导ROS生成激活hPDLCs自噬,保护细胞免受营养缺乏的损害。

关键词: 活性氧, 自噬, 饥饿, N-乙酰半胱氨酸

Abstract: Objective: To study the changes of autophagy levels of human periodontal ligament cells (hPDLCs) under starvation conditions, and to explore the role of reactive oxygen species (ROS) in the process of autophagy. Methods: hPDLCs were isolated and cultured. EBSS was applied to simulate a nutrient-deficient environment. Transmission electron microscopy was used to observe the production of autophagosomes to evaluate the autophagy level in hPDLCs. In order to explore the potential regulatory mechanism of autophagy activation under starvation conditions, hPDLCs were pretreated with antioxidants N-acetylcysteine (NAC) to inhibit the generation of ROS. DCFH-DA staining was used to detect the level of ROS, and the intensity of autophagy were monitored by RT-qPCR and Western blot which measure the expression of autophagy-related proteins LC3-Ⅱ, Beclin-1, and p62. Results: After EBSS starvation, the autophagy level increased. In addition, NAC partially reversed the autophagy levels in starved hPDLCs by inhibiting ROS production. Conclusion: ROS act as a signal molecule to mediate autophagy. Starvation activates autophagy in hPDLCs by inducing ROS generation which protects cells from nutritional deficiency.

Key words: reactive oxygen species, autophagy, starvation, N-acetylcysteine

何海燕, 余京泓, 伍宇婕, 张闵, 徐晓梅. 饥饿环境中活性氧激活人牙周膜细胞自噬的研究[J]. 口腔医学研究, 2022, 38(9): 877-881.

HE Haiyan, YU Jinghong, WU Yujie, Zhang Min, XU Xiaomei. A Study on Activation of Autophagy in Human Periodontal Ligament Cells by Reactive Oxygen Species in Starvation Environment.[J]. Journal of Oral Science Research, 2022, 38(9): 877-881.

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饥饿环境中活性氧激活人牙周膜细胞自噬的研究

A Study on Activation of Autophagy in Human Periodontal Ligament Cells by Reactive Oxygen Species in Starvation Environment.

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