| 摘要: |
| 目的 明确虎杖苷对芥子气所致肺损伤的改善作用，并初步探索其作用机制。方法 在存活率实验中，将雄性ICR小鼠随机分为6组（n＝15）：对照组，芥子气（40 mg/kg）组，虎杖苷低、中、高剂量（100、200、400 mg/kg，每天1次，持续7 d）组，以及阳性对照药（N-乙酰半胱氨酸200 mg/kg，每天1次，持续7 d）组；在存活率以外的实验中，将雄性ICR小鼠随机分为4组（n＝15）：对照组、芥子气（30 mg/kg）组、虎杖苷中剂量（200 mg/kg，每天1次，持续5 d）组、阳性对照药（N-乙酰半胱氨酸200 mg/kg，每天1次，持续5 d）组。皮下注射芥子气溶液建立小鼠芥子气肺损伤模型，虎杖苷处理组与阳性对照药组灌胃给药。通过存活率实验初步评价虎杖苷对芥子气致肺损伤的作用效果，确定虎杖苷最佳给药剂量；通过肺组织切片评价虎杖苷对芥子气致肺损伤的改善作用；通过ELISA法检测超氧化物歧化酶（SOD）、谷胱甘肽（GSH）、髓过氧化物酶（MPO）、丙二醛（MDA）和过氧化氢等氧化应激指标及IL-1β、IL-6和TNF-α等炎症反应指标的变化；通过蛋白质印迹法考察沉默信息调节因子1（SIRT1）、核因子E2相关因子2（Nrf2）、血红素加氧酶1（HO-1）和NAD （P） H：醌氧化还原酶1（NQO1）等氧化应激关键蛋白及Toll样受体4（TLR4）、NF-κB p65等炎症关键蛋白的表达水平。结果 与芥子气组相比，虎杖苷中、高剂量组小鼠的存活率升高，其中虎杖苷中剂量组效果更显著（P<0.01）。虎杖苷中剂量干预可以升高染毒小鼠肺组织中SOD活性、GSH含量，降低染毒小鼠肺组织中MPO活性、MDA含量、过氧化氢含量及IL-1β、IL-6和TNF-α等炎症因子的水平（P<0.05或P<0.01）；提升染毒小鼠肺组织中SIRT1蛋白的表达，促进Nrf2的核转移，提升HO-1、NQO1蛋白的表达（P<0.05或P<0.01）；降低染毒小鼠肺组织中TLR4、NF-κB p65总蛋白及磷酸化NF-κB p65蛋白的表达（P<0.01）。结论 虎杖苷可能通过调控SIRT1/Nrf2和TLR4/NF-κB通路抑制芥子气诱导的氧化应激和炎症反应，从而改善芥子气导致的小鼠肺损伤。 |
| 关键词: 芥子气 虎杖苷 氧化性应激 炎症反应 沉默信息调节因子1 核因子E2相关因子2 Toll样受体4 NF-κB |
| DOI：10.16781/j.CN31-2187/R.20211027 |
| 投稿时间：2021-10-13修订日期：2022-01-06 |
| 基金项目:国家自然科学基金（81871521，81801869），上海市公共卫生优秀青年人才项目（GWV-10.2-YQ18），上海市自然科学基金（21ZR1477700），上海市青年科技英才扬帆计划（21YF1457400），海军军医大学（第二军医大学）未来战争项目（19WLMS-08）. |
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| Protective effect of polydatin on sulfur mustard-induced lung injuries in mice |
| WANG Yu-run1,CEN Jin-feng2,MENG Wen-qi2,PEI Zhi-peng2,XIAO Kai2,XIE Ying1*,SUN Ming-xue2* |
(1. Key Laboratory of Molecular Epidemiology of Hunan Province, Medical College of Hunan Normal University, Changsha 410000, Hunan, China;
2. Department of Chemical Defense Medicine, Faculty of Naval Medicine, Naval Medical University(Second Military Medical University), Shanghai 200433, China
*Corresponding authors) |
| Abstract: |
| Objective To investigate the ameliorative effects and preliminary mechanism of polydatin on sulfur mustard (SM)-induced lung injuries. Methods In the survival experiment, male ICR mice were randomly divided into 6 groups (n=15):control group; SM group (40 mg/kg); low-, medium- and high-does polydatin groups (100, 200 and 400 mg/kg daily for 7 d, respectively); and positive control group (200 mg/kg N-acetyl-L-cysteine daily for 7 d). In other experiments, mice were randomly divided into 4 groups (n=15):control group, SM group (30 mg/kg), medium-dose polydatin group (200 mg/kg daily for 5 d) and positive control group (200 mg/kg N-acetyl-L-cysteine daily for 5 d). The mouse model of SM-induced lung injuries was established by subcutaneous injection of SM solution. The polydatin group and positive control group were given drugs by gavage. The survival experiment was used to preliminarily investigate the effects of polydatin on SM-induced lung injuries and to determine the best dose of polydatin. The ameliorative effect of polydatin on SM-induced lung injuries was evaluated by lung tissue sections. The levels of oxidative stress indexes such as superoxide dismutase (SOD), glutathione (GSH), myeloperoxidase (MPO), malondialdehyde (MDA) and hydrogen peroxide and inflammatory response indexes such as interleukin (IL)-1β, IL-6 and tumor necrosis factor α (TNF-α) were detected by enzyme-linked immunosorbent assay. The expression levels of key proteins of oxidative stress such as silencing information regulator 1 (SIRT1), nuclear factor E2-related factor 2 (Nrf2), heme oxygenase 1 (HO-1), NAD(P)H:quinone oxidoreductase 1 (NQO1), as well as the levels of key inflammatory proteins such as Toll-like receptor 4 (TLR4) and nuclear factor κB (NF-κB) p65 were detected by Western blotting. Results Compared with the SM group, the survival rates of mice were increased in the medium- and high-does polydatin groups, and the effect of the medium-dose polydatin group was more significant (P<0.01). Intervention with medium-dose polydatin could increase SOD activity and GSH content in the lung tissues of SM-exposed mice, and reduce MPO activity, contents of MDA and hydrogen peroxide, and the levels of inflammatory cytokines such as IL-1β, IL-6 and TNF-α (P<0.05 or P<0.01); enhance the protein expression of SIRT1 in lung tissues of SM-exposed mice, promote nuclear metastasis of Nrf2, and enhance the protein expression of HO-1 and NQO1 (P<0.05 or P<0.01); and decrease the protein expression of TLR4, NF-κB p65 and phosphorylated NF-κB p65 in lung tissues (P<0.01). Conclusion Polydatin may inhibit SM-induced oxidative stress and inflammatory response by regulating SIRT1/Nrf2 and TLR4/NF-κB pathways, so as to ameliorate the SM-induced lung injuries in mice. |
| Key words: sulfur mustard polydatin oxidative stress inflammation silencing information regulator 1 nuclear factor E2-related factor 2 Toll-like receptor 4 NF-κB |
