目的 基于磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)/哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)通路探讨麻黄-苦杏仁药对(Ephedrae Herba and Armeniacae Semen Amarum combination,EAC)通过调控巨噬细胞极化缓解急性肺损伤(acute lung injury,ALI)的作用机制。方法 采用高效液相色谱法(high-performance liquid chromatography,HPLC)检测EAC提取物中主要成分含量。结合网络药理学分析筛选EAC的潜在作用靶点及关键通路。建立脂多糖(lipopolysaccharide,LPS)诱导的ALI小鼠模型,给予EAC或地塞米松干预后,通过肺组织病理学、肺组织湿/干质量比、炎性因子水平检测,结合免疫荧光、流式细胞术、qRT-PCR及Western blotting评估EAC的抗炎效应及其对巨噬细胞极化和PI3K/Akt/mTOR信号通路的调控作用。结果 网络药理学结果显示,PI3K/Akt/mTOR通路为EAC治疗ALI的核心调控机制。动物实验结果显示,EAC呈剂量相关性地缓解ALI,显著降低ALI小鼠肺组织病理评分、肺组织湿/干质量比及血清中促炎因子水平(P＜0.05、0.01),显著升高血清中抗炎因子水平(P＜0.01)。机制研究结果显示,EAC可促进M2型巨噬细胞极化,抑制PI3K/Akt/mTOR通路异常激活,从而有效减轻肺部炎症反应。结论 EAC通过调节巨噬细胞M1/M2极化状态并抑制PI3K/Akt/mTOR信号通路,有效缓解LPS诱导的ALI炎症损伤。

Objective To explore the mechanism of Mahuang (Ephedra Herba) and Kuxingren (Armeniaca Semen Amarum) combination (EAC) in alleviating acute lung injury (ALI) by regulating macrophage polarization based on phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) pathway. Methods High performance liquid chromatography (HPLC) was used to detect the contents of major components in EAC extract. Potential targets and key pathways of EAC were screened through network pharmacology analysis. Lipopolysaccharide (LPS)-induced ALI mouse model was established, EAC or dexamethasone were administered for intervention, the anti-inflammatory effect of EAC and its regulatory effect on macrophage polarization and PI3K/Akt/mTOR signaling pathway were evaluated through detection of lung tissue pathology, lung tissue wet/dry weight ratio, inflammatory factor levels combined with immunofluorescence, flow cytometry, qRT-PCR and Western blotting. Results The results of network pharmacology showed that PI3K/Akt/mTOR pathway was the core regulatory mechanism of EAC in treatment of ALI. Animal experiment results showed that EAC dose dependently alleviated ALI, significantly reduced lung tissue pathological score, lung tissue wet/dry weight ratio and pro-inflammatory factors levels in serum of ALI mice (P < 0.05, 0.01), and significantly increased anti-inflammatory factor level in serum (P < 0.01). The mechanism research results showed that EAC could promote polarization of M2 macrophages, inhibit abnormal activation of PI3K/Akt/mTOR pathway, and effectively alleviate pulmonary inflammatory response. Conclusion EAC effectively alleviates LPS-induced ALI inflammatory damage by regulating macrophage M1/M2 polarization and inhibiting PI3K/Akt/mTOR signaling pathway.

R285.5

2022年银川市学术技术带头人储备工程项目