School of Pharmacy, Anhui University of Chinese Medicine, Hefei, Anhui, 230012, China.
School of Pharmacy, Anhui University of Chinese Medicine, Hefei, Anhui, 230012, China; Anhui Province Key Laboratory of Research & Development of Chinese Medicine, Hefei, Anhui, 230012, China.
Phytomedicine. 2023 Jun;114:154774. doi: 10.1016/j.phymed.2023.154774. Epub 2023 Mar 24.
Chronic obstructive pulmonary disease (COPD) is currently the third leading cause of death globally. Oxidative stress affects various molecular mechanisms and is the main driving factor of COPD. Ally isothiocyanate (AITC) is an effective component of Semen Sinapis Albae, which has favorable effects for the treatment of COPD, but its mechanism has not been fully elucidated.
This study aimed to elucidate the antioxidant effect of AITC on COPD and its molecular mechanism, and preliminarily determine the role of AhR in the progression of COPD.
The COPD rat model was established by smoking combined with intratracheal instillation of lipopolysaccharide. Different doses of AITC, positive control drug acetylcysteine, AhR inhibitor alpha-naphthoflavone, and agonist beta-naphthoflavone were administered by gavage. Human bronchial epithelial cells induced by cigarette smoke extract (CSE) were used in an in vitro model to explore the molecular mechanisms of AITC.
The effects of AITC on lung function and oxidative stress in rats were evaluated in vivo using the respiratory function test, white blood cell count, enzyme-linked immunosorbent assay, and histological staining. The changes in protein expression in the lung tissue were detected by immunohistochemistry and Western blotting. RT-PCR, western blotting, and immunofluorescence were used to explore the molecular mechanisms of AITC. Enzyme-linked immunosorbent assay, reactive oxygen species probing, and flow cytometry were used to determine the antioxidant effect of AITC.
AITC can improve the lung function of rats with COPD, restore lung tissue structure, improve oxidative stress, reduce inflammation, and inhibit lung cell apoptosis. AITC reversed the upregulation of AhR and CYP1A1 and the down-regulation of Nrf2 and NQO1 in the lung tissues of rats with COPD. CSE stimulation can increase the expressions of AhR and CYP1A1 and decrease the expressions of Nrf2 and NQO1 in 16HBE cells, leading to severe oxidative stress and inflammatory response and, ultimately, apoptosis. AITC inhibited AhR and CYP1A1 expressions, induced Nrf2 and NQO1 expressions, promoted Nrf2 nuclear translocation, and improved CSE-induced toxicological effects.
AITC may improve lung oxidative stress by inhibiting the AhR / CYP1A1 and activating the Nrf2 / NQO1 pathways, thereby delaying the pathological progression of COPD.
慢性阻塞性肺疾病（COPD）是目前全球范围内的第三大致死原因。氧化应激影响多种分子机制，是 COPD 的主要驱动因素。丙烯基异硫氰酸酯（AITC）是白芥子的有效成分，对 COPD 的治疗有良好的效果，但作用机制尚未完全阐明。
本研究旨在阐明 AITC 对 COPD 的抗氧化作用及其分子机制，并初步确定 AhR 在 COPD 进展中的作用。
通过吸烟联合气管内滴注脂多糖建立 COPD 大鼠模型。通过灌胃给予不同剂量的 AITC、阳性对照药乙酰半胱氨酸、AhR 抑制剂 α-萘黄酮和激动剂 β-萘黄酮。采用香烟烟雾提取物（CSE）诱导的人支气管上皮细胞建立体外模型，探讨 AITC 的分子机制。
通过呼吸功能测试、白细胞计数、酶联免疫吸附试验和组织学染色，在体内评估 AITC 对大鼠肺功能和氧化应激的影响。通过免疫组织化学和 Western blot 检测肺组织中蛋白表达的变化。采用 RT-PCR、Western blot 和免疫荧光法探讨 AITC 的分子机制。酶联免疫吸附试验、活性氧探针和流式细胞术测定 AITC 的抗氧化作用。
AITC 可改善 COPD 大鼠的肺功能，恢复肺组织结构，改善氧化应激，减轻炎症，抑制肺细胞凋亡。AITC 逆转了 COPD 大鼠肺组织中 AhR 和 CYP1A1 的上调以及 Nrf2 和 NQO1 的下调。CSE 刺激可增加 16HBE 细胞中 AhR 和 CYP1A1 的表达，降低 Nrf2 和 NQO1 的表达，导致严重的氧化应激和炎症反应，最终导致细胞凋亡。AITC 抑制 AhR 和 CYP1A1 的表达，诱导 Nrf2 和 NQO1 的表达，促进 Nrf2 核转位，改善 CSE 诱导的毒理学作用。
AITC 可能通过抑制 AhR/CYP1A1 并激活 Nrf2/NQO1 通路来改善肺组织的氧化应激，从而延缓 COPD 的病理进展。
