目的 探究艾纳香总黄酮干预慢性咽炎的作用及其可能机制。方法 利用网络药理学与分子对接分析艾纳香总黄酮干预慢性咽炎的生物调控过程与信号通路，构建蛋白质-蛋白质相互作用（ PPI）网络和“成分-靶点”网络。建立氨水诱导的慢性咽炎大鼠模型，给予艾纳香总黄酮干预后，采用苏木素-伊红（ HE）染色观察大鼠咽部组织病理变化；酶联免疫吸附法（ ELISA）检测大鼠血清中白细胞介素-6（ IL-6）、前列腺素E₂（ PGE₂）和白细胞介素-10（ IL-10）水平；实时荧光定量PCR（ qRT-PCR）与蛋白免疫印迹（ Western blotting）检测大鼠咽部组织Toll样受体4（ TLR4）/磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（ Akt）/核转录因子-κB（ NF-κB）信号通路上关键基因与蛋白的表达。结果 共获得药物与疾病交集靶点177个，网络药理学与分子对接结果显示艾纳香总黄酮可以调节IL6、磷脂酰肌醇3激酶催化亚单位α（PI3KCA）、蛋白激酶B1（ Akt1）、核因子κB1（ NF-κB1）等核心靶点，调节异源性刺激反应等生物学过程，调控PI3K/Akt信号通路，从而干预慢性咽炎。体内实验结果表明，艾纳香总黄酮能明显改善大鼠咽部组织病理状态；显著降低大鼠血清中IL-6和PGE₂水平（ P＜0.05、0.01），升高IL-10水平（ P＜0.01）；显著下调大鼠咽部组织TLR4、PI3KCA、磷脂酰肌醇-3激酶调节亚基1（ PI3KR1）、Akt1、核因子κB激酶亚基β抑制因子（ IKBKB）和NF-κB1 mRNA表达水平（ P＜0.01），TLR4、PI3K、Akt、核因子κB p65（ NF-κBp65）蛋白表达水平及其磷酸化水平（ P＜0.05、0.01）。结论 艾纳香总黄酮可能通过调控TLR4/PI3K/Akt/NF-κB信号通路，减轻炎症反应，从而改善慢性咽炎。

Objective To investigate the effects and mechanisms of total flavonoids from Blumea balsamifera in rats with chronic pharyngitis (CP). Methods The biological regulatory processes and signaling pathways of total flavonoids from B. balsamifera on improving CP were predicted by network pharmacology and molecular docking, protein-protein interaction (PPI) network and “component-targets” network were constructed. A rat model of CP induced by ammonium hydroxide was established, and after intervention with total flavonoids from B. balsamifera. Hematoxylin-eosin (HE) staining was used to observe pathological changes in pharyngeal tissue. Enzyme linked immunosorbent assay was used to detect the levels of interleukin-6 (IL-6), prostaglandin E₂ (PGE₂), IL^-10 in serum. Real-time PCR (qRT-PCR) and Western blotting were used to detect the expression of key genes and proteins in Tolllike receptor 4 (TLR4)/phospholipinositide 3-kinase (PI3K)/protein kinase B (Akt)/nuclear transcription factor-κB (NF-κB) signaling pathway in rat pharyngeal tissue. Results A total of 177 interaction targets between active ingredients and diseases. The results of network pharmacology and molecules docking showed that total flavonoids from B. balsamifera could regulate core targets such as IL6, phosphatidylinositol 3-kinase catalytic subunit alpha (PI3KCA), protein kinase B1(Akt1), nuclear factor kappa B1(NF-κB1), regulate biological processes such as response to xenobiotic stimulus, and regulate PI3K/Akt signaling pathway, intervening in CP. The in vivo experimental results showed that the total flavones of B. balsamifera could improve the damage of pharyngeal tissue in rats with CP, significantly decreased the levels of IL-6, PGE₂ (P < 0.05, 0.01), increased IL-10 (P < 0.01) in serum significantly downregulate the expressions of TLR4, PI3KCA, phosphoinositide-3-kinase regulatory subunit1 (PI3KR1), Akt1, Inhibitor of nuclear factor kappa B kinase subunit beta (IKBKB) and NF-κB1 mRNA (P < 0.05, 0.01), TLR4, PI3K, Akt, nucler factor kappa B p65 (NF-κBp65) protein expressions and phosphorylation levels (P < 0.05, 0.01). Conclusion The total flavonoids from B. balsamifera propably improve CP by inhibiting TLR4/PI3K/Akt/NF-κB signaling pathway, suppressing inflammatory responses.

R285.5

国家中医药管理局高水平中医药重点学科建设项目（zyyzdxk-2023186）；2023年贵州“黔六味”道地药材优势特色产业集群建设项目“贵州道地药材种质资源库设备建设”；贵州中医药大学喀斯特药用资源保护与创新利用科技创新人才团队项目