氧化应激致自噬在光气诱导的急性肺损伤中的作用

doi:10.3969/j.issn.1004-616x.2023.05.001

摘要/Abstract

摘要： 目的： 探讨大鼠光气暴露模型中肺组织氧化水平的变化，以及氧化应激是否通过调控自噬缓解光气诱导的大鼠急性肺损伤(ALI)。方法： 将50只大鼠分为对照组、光气染毒后6、12、24和48 h组。对照组大鼠鼻吸入空气，光气染毒组大鼠鼻吸入41mg/m³的光气，动态染毒30 min。检测相关指标后，选择染毒后12 h组作为本研究动物模型(后称光气组)。另将40只大鼠分为对照组(腹腔注射0.9%生理盐水)、光气组(41 mg/m³的光气染毒30 min)、RPM处理组(光气染毒后腹腔注射5 mg/kg的RPM)和RPM对照组(腹腔注射5 mg/kg的RPM)。再取40只大鼠分为对照组(腹腔注射0.9%生理盐水)、光气组(41 mg/m³的光气染毒30 min)、NAC处理组(光气染毒后腹腔注射200 mg/kg的NAC)和NAC对照组(腹腔注射200 mg/kg的NAC)。观察大鼠肺组织病理学改变；检测大鼠肺功能、肺湿质量、肺系数和肺泡灌洗液(BALF)总蛋白浓度；测定大鼠肺组织ROS水平、肺组织脂质过氧化水平、抗氧化酶(SOD、CAT)活性以及GSH含量；检测氧化还原及自噬相关分子mRNA和蛋白表达水平。结果： 与对照组相比，大鼠光气染毒后不同时间组织病理学观察可见肺泡融合，炎性细胞增多；肺功能明显下降(P<0.05或0.01)；肺湿质量、肺系数和BALF总蛋白浓度显著升高(P<0.05 或 0.01)；大鼠肺组织中 ROS 和脂质过氧化水平升高(P<0.01)；各抗氧化酶活性和表达水平发生变化(P<0.05 或0.01)，以暴露后12 h组变化最为显著；与对照组相比，光气组自噬相关蛋白Beclin-1、LC3II/LC3I表达水平降低，P62表达水平上升(P<0.05或0.01)；与光气组相比，RPM处理组和NAC处理组自噬水平上调(P<0.05)并缓解光气诱导的ALI；同时，NAC处理组降低了肺组织ROS水平(P<0.05或P<0.01)。结论： 光气染毒后不同时间点大鼠肺组织氧化水平升高，同时氧化应激能够调控自噬水平，给予NAC可缓解氧化应激，从而上调自噬水平，改善光气所致的大鼠ALI。

关键词: 光气, 急性肺损伤, 氧化应激, 自噬

Abstract: OBJECTIVE: To explore changes of oxidation levels in lung tissues among the phosgeneexposedrats,and to investigate if oxidative stress would regulate autophagy to alleviate phosgene-induced acute lung injury (ALI) in rats. METHODS: Fifty rats were divided into five groups:control and treatment groups with phosgene treatment for 30 minutes and evaluated6,12,24 and 48 h later. The rats in the control group inhaled air through their noses, and the rats in the phosgene exposure group inhaled 41 mg/m³ of phosgene through their noses. After detecting relevant indicators,the 12 h post-exposure group was selected as the animal model for this study (hereinafter referred to as the phosgene group). Another 40 rats were divided into a control group (intraperitoneal injection of 0.9% normal saline),a phosgene group (41 mg/m³ of phosgene exposure for 30 min),and an RPM treatment group (intraperitoneal injection of 5 mg/kg of RPM after exposure) and the RPM control group (intraperitoneal injection of 5 mg/kg of RPM). Another 40 rats were divided into the control group (intraperitoneal injection of 0.9% normal saline), the phosgene group (41 mg/m³ of phosgene exposure for 30 min),and the NAC treatment group (the intraperitoneal injection of 200 mg/kg of NAC after exposure) and NAC control group (intraperitoneal injection of 200 mg/kg of NAC). These rats were observed for pathological changes in lung tissues; lung functions, lung moisture mass, lung coefficient, total protein concentration of bronchoalveolar lavage fluid (BALF), rat lung tissue ROS levels, lung tissue lipid peroxidation levels, anti-Oxidase (SOD, CAT) activity and GSH content, and redox and autophagy-related molecule mRNA and protein expression levels. RESULTS: Compared with the control group,histopathological observation at different time points after phosgene exposure showed alveolar fusion and increased inflammatory cells. Lung function decreased significantly (P<0.05 or 0.01). The lung moisture mass, lung coefficient and BALF total protein concentration were significantly increased (P<0.05 or 0.01). The levels of ROS and lipid peroxidation in lung tissues of rats were increased (P<0.01). The activity and expression levels of a series of antioxidant enzymes were changed (P<0.05 or 0.01),and the changes were most significant at 12 h after exposure. Compared with the control group, expression levels of autophagy-associated proteins, Beclin-1 and LC3II/LC3I, decreased in phosgene group,while the expression levels of P62 increased (P<0.05 or 0.01). Compared with phosgene group, the autophagy levels in RPM and NAC groups were up-regulated (P<0.05) and the phosgene induced ALI was alleviated. At the same time, the ROS level of lung tissue was decreased in the NAC treatment group. CONCLUSION: Oxidation levels of lung tissues increased at different time points after phosgene exposure. At the same time, oxidative stress regulated the level of autophagy. Administration of NAC alleviated oxidative stress,thereby upregulated autophagy levels and improved phosgene-induced ALI in rats.

Key words: phosgene, acute lung injury, oxidative stress, autophagy

中图分类号:

郭晓洁, 刘鹏晖, 龙子, 李佳威, 涂永梅, 李文丽, 刘启玲. 氧化应激致自噬在光气诱导的急性肺损伤中的作用[J]. 癌变·畸变·突变, 2023, 35(5): 325-333,340.

GUO Xiaojie, LIU Penghui, LONG Zi, LI Jiawei, TU Yongmei, LI Wenli, LIU Qiling. Induction of autophagy by oxidative stress in phosgene-induced acute lung injury[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2023, 35(5): 325-333,340.

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