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,南昌 330006;2.江西中医药大学附属医院,南昌 330006

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25

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R-33


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

1. School of Clinical Medicine, Jiangxi University of Traditional Chinese Medicine, Nanchang 330006, China.2. Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine, Nanchang 330006

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    摘要:

    目的 探讨蠲哮汤(Juanxiao decotion, JXD)调节三型固有淋巴样细胞( type 3 innate lymphoid cells,ILC3s)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)、JXD 组(低、中、高剂量分别为 8. 5、17、34 g / kg)、地塞米松组(1 mg / kg),每组 10 只。 除正常组,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,JXD 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 7 d。 苏木素-伊红( hematoxylin-eosin,HE)染色观察肺部病理改变,全自动生化仪检测血脂四项及肺泡灌洗液( alveolar lavage fluid,BALF)炎症细胞分类计数,酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测 BALF 和血清免疫球蛋白 E(immunoglobulin E,IgE)水平,肺组织中白细胞介素- 1β( interleukin-1β,IL-1β)、白细胞介素- 13( interleukin-13,IL-13)、C-C 趋化因子受体 17(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 STAT3 蛋白磷酸化水平。 结果 与正常组相比,模型组小鼠气道炎性细胞浸润,气道壁增厚,经地塞米松和 JXD 中、高剂量干预后小鼠肺部炎症情况得到改善,其中高剂量效果更明显。 与正常组相比,模型组小鼠肺组织中 IL-1β、IL-17A+ILC3、IL-13、CCL17 水平显著升高(P<0. 05),IL-22+ILC3 比例和 P-STAT3 蛋白表达水平明显降低( P<0. 01,P<0. 05);与模型组相比,经 JXD 中、高剂量干预后肺组织中 IL-1β、IL-17A+ILC3、IL-13、CCL17 水平显著下降,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 JXD 能改善肥胖哮喘小鼠机体炎症环境,减轻肺部炎症和过敏反应,其机制可能与调节 ILC3 分泌细胞因子功能有关。

    Abstract:

    Objective To explore the mechanism of Juanxiao decoction in regulating type 3 innate lymphoid cells( ILC3s) in treating obese asthma. Methods Sixty male BALB/ c mice were randomly divided into a normal group, model group ( high-fat diet + OVA), Juanxiao decoction groups ( low, middle, and high doses of 8. 5, 17, and 34 g / kg,respectively), and dexamethasone group (1 mg / kg) with 10 mice in each group. Except for the normal group, the other groups were fed a high-fat diet for 12 weeks, and OVA sensitization by inhalation of an atomized OVA solution was used to establish the obese asthma model. From the first inhalation, the low-, medium-, and high-dose groups of Juanxiao decoction and the dexamethasone group were administered corresponding drugs by gavage, whereas normal and model groups were administered equal amounts of saline by gavage for 7 days. The state of mice and changes in typical symptoms of obese asthma were observed. At 24 hours after the last challenge, a fully automated biochemical analyzer was used to assess four blood lipids and count inflammatory cells in alveolar lavage fluid (BALF). Hematoxylin-eosin staining was used to observe morphological changes in lung tissue and abdominal fat. Enzyme-linked immunosorbent assays were used to measure the immunoglobulin E in BALF and serum, and interleukin ( IL)-1β, IL-13, and mouse thymus activation regulating chemokine (CCL17) in lung tissue. IL-17A+ILC3 and IL-22+ILC3 in lung tissue and peripheral blood were analyzed by flow cytometry. Western blot was used to detect expression of P-STAT3 protein in lung tissue. Results Compared with the normal group, model group mice showed infiltration of airway inflammatory cells and thickening of airway walls. However, compared with the model group, lung inflammation in dexamethasone and Juanxiao decoction groups was improved, especially in middle- and high-dose groups. Compared with the normal group, IL-1β, IL-17A+ILC3, IL-13,and CCL17 in lung tissue of the model group were significantly increased (P<0. 05), whereas the proportion of IL-22+ILC3 and expression of P-STAT3 were significantly decreased (P<0. 01, P<0. 05). Compared with the model group, IL1β, IL-17A+ILC3, IL-13, and CCL17 in lung tissue were significantly decreased and the proportion of IL-22+ILC3 andexpression of P-STAT3 were significantly increased in middle- and high-dose Juanxiao decoction groups ( P<0. 05, P<0. 01, P<0. 001). Conclusions Juanxiao decoction improves the inflammatory environment of obese asthmatic mice and alleviates lung inflammatory and allergic reactions. Its mechanism may be related to regulating secretion of cytokines by ILC3s.

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田敏萍,张清源,向双娣,陈玲玲,孙 朋,薛汉荣.蠲哮汤调节三型固有淋巴样细胞治疗肥胖哮喘小鼠机制[J].中国比较医学杂志,2024,34(5):~13.

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  • 收稿日期:2023-09-25
  • 在线发布日期: 2024-06-26
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