目的 探究2,6-二甲氧基-1,4-苯醌（2,6-dimethoxy-1,4-benzoquinone, 2,6-DMBQ）在哮喘小鼠中的治疗作用及其机制。 方法 将SPF级BALB/c小鼠随机分为正常对照组、卵清蛋白（ovalbumin, OVA）组、融媒组、布地奈德（budesonide, BUD）组、2,6-DMBQ低、中、高剂量组，每组8只。采用OVA诱导法建立哮喘小鼠模型并给予相应药物干预。使用无创肺功能仪测定小鼠气道高反应性，ELISA法测定支气管肺泡灌洗液中白细胞介素（interleukin, IL）-17、IL-10和血清免疫球蛋白E水平，细胞计数仪检测支气管肺泡灌洗液中嗜酸性粒细胞数，苏木精-伊红染色和过碘酸-希夫染色评估肺组织病理变化，Western blot检测肺组织哺乳动物雷帕霉素靶蛋白通路相关蛋白（p-AKT/AKT、p-p70S6K/p70S6K）表达情况，全自动生化分析仪测定肝肾功能。 结果 与正常对照组相比，OVA组增强呼吸间歇值、苏木精-伊红染色炎性评分和过碘酸-希夫染色阳性面积、嗜酸性粒细胞百分比、IL-17/IL-10比值、血清免疫球蛋白E水平、p-AKT/AKT和p-p70S6K/p70S6K蛋白相对表达量升高（P<0.05）；与OVA组相比，BUD组和2,6-DMBQ中、高剂量组上述指标降低（P<0.05）。 结论 2,6-DMBQ可抑制哺乳动物雷帕霉素靶蛋白通路减轻哮喘小鼠气道炎症，可能通过减轻辅助性T细胞17/调节性T细胞失衡发挥作用。
Abstract
Objective To investigate the therapeutic effects and mechanisms of 2,6-dimethoxy-1,4-benzoquinone (2,6-DMBQ) in a mouse model of asthma. Methods SPF-grade BALB/c mice were randomly divided into 7 groups (n=8 each group): normal control group, ovalbumin (OVA) group, dimethyl sulfoxide+corn oil group, budesonide (BUD) group, and low, medium, and high dose 2,6-DMBQ groups. An asthma mouse model was established by OVA induction, followed by corresponding drug interventions. Non-invasive lung function tests were performed to measure airway hyperresponsiveness, and enzyme-linked immunosorbent assay was used to determine levels of interleukin (IL)-17, IL-10, and serum immunoglobulin E in bronchoalveolar lavage fluid. A cell counter was employed to detect eosinophil counts in bronchoalveolar lavage fluid, while hematoxylin-eosin staining and periodic acid-Schiff staining were used to assess lung tissue pathological changes. Western blot was conducted to examine the expression of proteins related to the mammalian target of rapamycin pathway (p-AKT/AKT and p-p70S6K/p70S6K), and a fully automated biochemical analyzer was used to evaluate liver and kidney functions. Results Compared with the normal control group, the OVA group showed increased enhanced pause values, inflammation scores from hematoxylin-eosin staining, positive area from periodic acid-Schiff staining, percentage of eosinophils, IL-17/IL-10 ratio, serum immunoglobulin E levels, and relative expression levels of p-AKT/AKT and p-p70S6K/p70S6K (P<0.05). The BUD group and the medium and high dose 2,6-DMBQ groups exhibited decreased values for these indicators compared to the OVA group (P<0.05). Conclusions 2,6-DMBQ can inhibit the mTOR pathway to alleviate airway inflammation in asthmatic mice, possibly by mitigating the imbalance between Th17 and regulatory T cells.
关键词
支气管哮喘 /
2 /
6-二甲氧基-1 /
4-苯醌 /
哺乳动物雷帕霉素靶蛋白通路 /
Th17/Treg失衡 /
小鼠
Key words
Bronchial asthma /
2,6-Dimethoxy-1,4-benzoquinone /
Mammalian target of rapamycin pathway /
Th17/Treg imbalance /
Mouse
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基金
河南省重点研发与推广专项（科技攻关）（232102310323）。
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