实验动物与比较医学 ›› 2026, Vol. 46 ›› Issue (2): 191-204.DOI: 10.12300/j.issn.1674-5817.2025.084

• 人类疾病动物模型 • 上一篇    下一篇

泻白散对过敏性哮喘大鼠肺、肠组织形态结构及PI3K和Akt表达水平的影响

宋静1,2,3, 杨宗统2()(), 李晓晶2, 李自发1, 苏凤云3, 徐东川2, 隋在云2()()   

  1. 1.山东中医药大学, 济南 250355
    2.山东省中医药研究院中药药理研究所, 济南 250014
    3.山东第一医科大学第二附属医院, 泰安 271000
  • 收稿日期:2025-06-10 修回日期:2025-08-21 出版日期:2026-04-25 发布日期:2026-04-18
  • 通讯作者:
    杨宗统(1992—),男,博士,助理研究员,研究方向:中药免疫药理及药效学。E-mail: YangZT2021@163.com。ORCID:0009-0005-5401-7697
    隋在云(1964—),女,硕士生导师,研究员,研究方向:中药及其有效成分作用机制及安全性评价。E-mail: zy_505@163.com。
    ORCID:0009-0008-4487-2317
  • 作者简介:宋 静(1987—),女,硕士研究生,研究方向:中药及其有效成分作用机制及安全性评价。E-mail:583023072@qq.com
  • 基金资助:
    山东省重大科技创新工程项目“基于网络药理学技术对泻白散药效成分-哮喘靶标关联性的探索研究”(2018CXGC1301);山东省中医药科技项目“基于PI3K-Akt信号通路探讨经典名方泻白散治疗过敏性哮喘的肠道黏膜免疫作用机制”(2021Q126);山东省中医药研究院科研孵化基金项目“基于PI3K-Akt信号通路的过敏性哮喘大鼠‘肺与大肠相表里’免疫机制研究”(2021SACM-1)

Effects of Xiebai San on the Morphological Structures of Lung and Intestinal Tissues and Expression Levels of PI3K and Akt in Rats with Allergic Asthma

SONG Jing1,2,3, YANG Zongtong2()(), LI Xiaojing2, LI Zifa1, SU Fengyun3, XU Dongchuan2, SUI Zaiyun2()()   

  1. 1.Shandong University of Traditional Chinese Medicine, Jinan 250355, China
    2.Institute of Pharmacology of Traditional Chinese Medicine, Shandong Academy of Traditional Chinese Medicine, Jinan 250014, China
    3.The Second Affiliated Hospital of Shandong First Medical University, Tai'an 271000, China
  • Received:2025-06-10 Revised:2025-08-21 Published:2026-04-25 Online:2026-04-18
  • Contact: YANG Zongtong (ORCID: 0009-0005-5401-7697), E-mail: YangZT2021@163.com
    SUI Zaiyun (ORCID: 0009-0008-4487-2317), E-mail: zy_505@163.com

摘要:

目的 探讨泻白散对过敏性哮喘大鼠呼吸道和肠道黏膜免疫的作用机制。 方法 40只雄性SD大鼠按体重随机分为空白组、模型组、阳性药组和泻白散组,每组10只。模型组、阳性药组和泻白散组均使用卵清蛋白致敏法建立大鼠过敏性哮喘模型。自雾化激发期(第21天)开始,各组同时进行灌胃干预,即阳性药组大鼠灌胃地塞米松(0.068 mg/kg),泻白散组大鼠灌胃泻白散药液(2 g/mL,11.3 mL/kg),空白组与模型组大鼠灌胃等体积生理盐水,每日1次,连续14 d。将大鼠安乐死后,采集各组大鼠肺、肠组织,经HE染色观察组织病理学变化;透射电子显微镜观察组织超微病理结构;免疫组织化学法观察磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)和蛋白激酶B(protein kinase B,Akt)蛋白阳性面积。提取大鼠肺、肠组织中的总蛋白和总RNA,采用蛋白质印迹法和实时荧光定量PCR法分别检测PI3KAkt基因的蛋白和mRNA表达水平。 结果 组织病理学结果显示,与空白组相比,模型组大鼠出现肺泡性肺气肿伴炎性细胞浸润,以及肠黏膜损伤伴炎性细胞浸润;肺组织细胞结构被破坏,细胞器数量减少,但肠超微结构病变较轻。与模型组相比,泻白散组大鼠肺组织病变程度较低,偶见肺泡壁损伤伴少量炎性细胞浸润,肠黏膜结构改善,腺体排列整齐,疏松、浸润等病理改变均见好转;肺组织细胞结构比较完整,病变程度减轻,肠组织无超微结构病理变化。免疫组织化学和蛋白质印迹结果显示,与空白组相比,模型组大鼠肺、肠组织中PI3K和Akt特异性阳性反应面积均显著增大(均P<0.001),PI3K和Akt蛋白表达量也均显著增加(均P<0.05);与模型组相比,泻白散组大鼠肺组织中Akt蛋白阳性面积显著减少(P<0.001),肠组织中PI3K阳性面积显著减少(P<0.000 1),同时肺、肠组织中PI3K和Akt蛋白表达量均显著减少(均P<0.01)。实时荧光定量PCR结果显示,与空白组相比,模型组大鼠肺、肠组织中PI3KAkt基因的mRNA表达量均显著增加(均P<0.05);与模型组相比,泻白散组大鼠肺、肠组织中PI3KAkt基因的mRNA表达量均显著减少(均P<0.05)。 结论 泻白散可通过抑制过敏性哮喘大鼠肺、肠组织中PI3K-Akt信号通路关键核酸和蛋白表达,改善肺组织形态结构,保护肠道黏膜完整性,并调节肠道黏膜免疫功能等机制,发挥对过敏性哮喘大鼠的保护作用。

关键词: 泻白散, 哮喘, 黏膜免疫, PI3K-Akt信号通路, 大鼠

Abstract:

Objective To investigate the mechanism by which Xiebai San regulates respiratory tract and intestinal mucosal immunity in rats with allergic asthma. Methods Forty male SD rats were randomly divided into four groups based on body weight: control group, model group, positive control group, and Xiebai San group. The model group, positive control group, and Xiebai San group were sensitized with ovalbumin to establish a rat model of allergic asthma. From day 21 (the aerosol challenge phase), each group received daily gavage interventions simultaneously: the positive control group was administered dexamethasone (0.068 mg/kg), the Xiebai San group received Xiebai San solution (2 g/mL, 11.3 mL/kg), while the control and model groups were given an equal volume of normal saline, once daily for 14 consecutive days. After euthanasia, lung and intestinal tissues were collected. Hematoxylin and eosin staining was used to observe histopathological changes. Transmission electron microscopy was employed to examine tissue ultrastructure. Immunohistochemistry was applied to detect the positive reaction areas of phosphatidyl-inositol 3-kinase (PI3K) and protein kinase B (Akt) proteins. Total protein and total RNA were extracted from lung and intestinal tissues, then the protein and mRNA expression levels of PI3K and Akt genes were detected by Western blotting and real-time quantitative PCR, respectively. Results Histopathological results showed alveolar emphysema accompanied by inflammatory cell infiltration, and intestinal mucosal injury with inflammatory cell infiltration in the model group as compared with the control group; the cellular structure of lung tissues was disrupted in the model group, with reduced organelles, while the ultrastructural lesions in the intestine were relatively mild. Compared with the model group, Xiebai San group exhibited milder pathological changes in lung tissues, with occasional alveolar wall damage and a small amount of inflammatory cell infiltration; the intestinal mucosal structure was improved, glands were arranged regularly, and pathological changes such as tissue loosening and inflammatory infiltration were alleviated; the cellular structure of lung tissues was relatively intact with reduced severity of lesions, and no ultrastructural pathological changes were observed in intestinal tissues. Immunohistochemistry and Western blotting results showed that compared with the control group, the specific positive reaction areas of PI3K and Akt in lung and intestinal tissues were significantly increased in the model group (all P<0.001); meanwhile, the protein expression levels of PI3K and Akt were significantly upregulated (all P<0.05). Compared with the model group, the positive area of Akt protein in lung tissue was significantly reduced in the Xiebai San group (P<0.001), and the positive area of PI3K in intestinal tissue was also significantly decreased (P<0.000 1). Additionally, the protein expression levels of PI3K and Akt in lung and intestinal tissues were significantly downregulated (all P<0.01). Real-time quantitative PCR results showed that compared with the control group, the mRNA expression levels of PI3K and Akt genes in lung and intestinal tissues were significantly elevated in the model group (all P<0.05). Compared with the model group, the mRNA expression levels of PI3K and Akt genes in lung and intestinal tissues were significantly reduced in the Xiebai San group (all P<0.05). Conclusion Xiebai San exerts protective effects on rats with allergic asthma by inhibiting the expression of key nucleic acids and proteins in the PI3K-Akt signaling pathway in lung and intestinal tissues, improving the morphological structure of lung tissue, and maintaining intestinal mucosal integrity, and regulating intestinal mucosal immune function.

Key words: Xiebai San, Asthma, Mucosal immunity, PI3K-Akt signaling pathway, Rats

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