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)/哺乳動物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)通路探討麻黃-苦杏仁藥對(Ephedrae Herba and Armeniacae Semen Amarum combination,EAC)通過調(diào)控巨噬細(xì)胞極化緩解急性肺損傷(acute lung injury,ALI)的作用機制。方法 采用高效液相色譜法(high-performance liquid chromatography,HPLC)檢測EAC提取物中主要成分含量。結(jié)合網(wǎng)絡(luò)藥理學(xué)分析篩選EAC的潛在作用靶點及關(guān)鍵通路。建立脂多糖(lipopolysaccharide,LPS)誘導(dǎo)的ALI小鼠模型,給予EAC或地塞米松干預(yù)后,通過肺組織病理學(xué)、肺組織濕/干質(zhì)量比、炎性因子水平檢測,結(jié)合免疫熒光、流式細(xì)胞術(shù)、qRT-PCR及Western blotting評估EAC的抗炎效應(yīng)及其對巨噬細(xì)胞極化和PI3K/Akt/mTOR信號通路的調(diào)控作用。結(jié)果 網(wǎng)絡(luò)藥理學(xué)結(jié)果顯示,PI3K/Akt/mTOR通路為EAC治療ALI的核心調(diào)控機制。動物實驗結(jié)果顯示,EAC呈劑量相關(guān)性地緩解ALI,顯著降低ALI小鼠肺組織病理評分、肺組織濕/干質(zhì)量比及血清中促炎因子水平(P＜0.05、0.01),顯著升高血清中抗炎因子水平(P＜0.01)。機制研究結(jié)果顯示,EAC可促進M2型巨噬細(xì)胞極化,抑制PI3K/Akt/mTOR通路異常激活,從而有效減輕肺部炎癥反應(yīng)。結(jié)論 EAC通過調(diào)節(jié)巨噬細(xì)胞M1/M2極化狀態(tài)并抑制PI3K/Akt/mTOR信號通路,有效緩解LPS誘導(dǎo)的ALI炎癥損傷。

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年銀川市學(xué)術(shù)技術(shù)帶頭人儲備工程項目