蠲哮汤调节三型固有淋巴样细胞治疗肥胖哮喘小鼠机制
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1.江西中医药大学临床医学院;2.江西中医药大学附属医院

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国家自然科学基金项目(面上项目,重点项目,重大项目)


Mechanism of Juanxiao decoction regulating type 3 innate lymphoid cells in treating obese asthmatic mice
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Affiliation:

1.Jiangxi University of Chinese Medicine,Jiangxi Nanchang;2.Affiliated Hospital of Jiangxi University of Chinese Medicine

Fund Project:

The National Natural Science Foundation of China (General Program, Key Program, Major Research Plan)

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    摘要:

    摘要:目的:探讨蠲哮汤调节三型固有淋巴样细胞(ILC3)治疗肥胖型哮喘的可能作用机制。方法:60只BALB/C小鼠随机分为正常组、模型组(60%高脂饲料+OVA)、蠲哮汤组(低、中、高剂量分别为8.5、17、34 g?kg-1),醋酸地塞米松组(1 mg?kg-1),每组10只。除空白组,其余组高脂饲料喂养12周后,OVA致敏及雾化激发建立肥胖哮喘模型。首次雾化起,蠲哮汤低、中、高剂量组及地塞米松组给予相应药物灌胃,正常组和模型组给予等量生理盐水灌胃,持续7d。观察分析小鼠的一般状态及肥胖哮喘典型症状改变,HE染色观察肺部病理改变,全自动生化仪检测血脂四项和肺泡灌洗液(BALF)炎症细胞分类计数,ELISA检测BALF和血清IgE水平,肺组织中IL-1β、IL-13、CCL17水平,流式细胞术分析肺和外周血中IL-17A+ILC3、IL-22+ILC3水平,Western blot 检测肺组织中P-STAT3蛋白表达水平。结果:蠲哮汤中、高剂量组小鼠的一般状态和典型肥胖哮喘症状较模型组得到明显改善。模型组小鼠气道炎性细胞浸润,气道壁增厚,经地塞米松和蠲哮汤中、高剂量干预后小鼠肺部炎症情况明显改善,其中高剂量效果更明显。模型组小鼠肺组织中IL-1β、IL-17A+ILC3、IL-13、CCL17水平高于正常组(p<0.05),IL-22+ILC3比例和P-STAT3表达水平明显低于正常组(p <0.01,p<0.05),经蠲哮汤中、高剂量干预后IL-1β、IL-17A+ILC3、IL-13、CCL17水平显著下降(p<0.05, p<0.01,p<0.001),IL-22+ILC3比例和P-STAT3表达明显增加(p<0.05,p<0.01,p<0.001)。结论:蠲哮汤减轻肥胖哮喘小鼠肺部炎症和过敏反应,改善机体炎症环境,其机制可能与调节ILC3分泌细胞因子功能有关。

    Abstract:

    Objective: Explore the mechanism of Juanxiao Decoction in regulating the type 3 innate lymphoid cells (ILC3s) in the treatment of obese asthma. Method: Sixty male BALB/C mice were randomly divided into a normal group, a model group (High Fat Diet+OVA), and Juanxiao Decoction groups (low-, middle-, and high doses of 8.5, 17, and 34 g?kg-1), dexamethasone group (1 mg?kg-1), with10 in each group. Except for the normal group, other groups were fed high-fat diet for 12 weeks, and OVA sensitization and atomization inhalation were used to establish an obesity asthma model. From the first atomization, the low-, medium-, and high dose groups of Juanxiao decoction and the dexamethasone group were given corresponding drugs by gavage, while the normal group and model group were given equal amounts of saline by gavage for 7 days. Observing the state of each group of mice and changes in typical symptoms of obesity asthma. Within 24 hours after the last challenge, a fully automated biochemical analyzer was used to detect the four items of blood lipids and the classified count of inflammatory cells in alveolar lavage fluid (BALF). Hematoxylin- eosin (HE) staining was used to observe the morphological changes of lung tissue and abdominal fat in mice. Enzyme linked immunosorbent assay (ELISA) for detecting levels of immunoglobulin E (IgE) in BALF and serum of mice, as well as the levels of interleukin-1β(IL-1β)、Interleukin-13(IL-13)and mouse thymus activation regulating chemokine (CCL17) in lung tissue. The levels of IL-17A+ILC3 and IL-22+ILC3 cells in lung tissue and peripheral blood of mice were analyzed by flow cytometry. Western blot was used to detect the expression of P-STAT3 protein in lung tissue. Results: Compared with the model group, the general state and typical obese asthma symptoms in the middle and high dose groups of Juanxiao decoction were obviously improved. Pathological staining showed that the airway inflammatory cells infiltrated and the airway wall thickened in the model group. However, compared with the model group, the lung inflammation in dexamethasone group and Juanxiao decoction group was improved, especially in the middle and high dose groups. The levels of IL-1β, IL-17A+ILC3, IL-13 and CCL17 in lung tissue of mice in model group were higher than those in normal group (p<0.05), while the proportion of IL-22ILC3 and the expression of P-STAT3 were significantly lower than those in normal group(p <0.01,p<0.05). After the intervention of middle and high dose of Juanxiao decoction, the levels of IL-1β, 17A+ILC3, IL-13 and CCL17 decreased significantly(p<0.05,p<0.01,p<0.001)and the the proportion of IL-22ILC3 and the expression of P-STAT3 increased significantly(p<0.05,p<0.01,p<0.001).Conclusion: Juanxiao decoction can alleviate lung inflammation and allergic reaction in obese asthmatic mice, and improve the inflammatory environment of the body, and its mechanism may be related to regulating the secretion of cytokines by ILC3.

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  • 收稿日期:2023-09-25
  • 最后修改日期:2023-10-31
  • 录用日期:2024-05-21
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