目的 探究艾納香總黃酮干預(yù)慢性咽炎的作用及其可能機(jī)制。方法 利用網(wǎng)絡(luò)藥理學(xué)與分子對接分析艾納香總黃酮干預(yù)慢性咽炎的生物調(diào)控過程與信號通路，構(gòu)建蛋白質(zhì)-蛋白質(zhì)相互作用（ PPI）網(wǎng)絡(luò)和“成分-靶點(diǎn)”網(wǎng)絡(luò)。建立氨水誘導(dǎo)的慢性咽炎大鼠模型，給予艾納香總黃酮干預(yù)后，采用蘇木素-伊紅（ HE）染色觀察大鼠咽部組織病理變化；酶聯(lián)免疫吸附法（ ELISA）檢測大鼠血清中白細(xì)胞介素-6（ IL-6）、前列腺素E₂（ PGE₂）和白細(xì)胞介素-10（ IL-10）水平；實(shí)時(shí)熒光定量PCR（ qRT-PCR）與蛋白免疫印跡（ Western blotting）檢測大鼠咽部組織Toll樣受體4（ TLR4）/磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（ Akt）/核轉(zhuǎn)錄因子-κB（ NF-κB）信號通路上關(guān)鍵基因與蛋白的表達(dá)。結(jié)果 共獲得藥物與疾病交集靶點(diǎn)177個，網(wǎng)絡(luò)藥理學(xué)與分子對接結(jié)果顯示艾納香總黃酮可以調(diào)節(jié)IL6、磷脂酰肌醇3激酶催化亞單位α（PI3KCA）、蛋白激酶B1（ Akt1）、核因子κB1（ NF-κB1）等核心靶點(diǎn)，調(diào)節(jié)異源性刺激反應(yīng)等生物學(xué)過程，調(diào)控PI3K/Akt信號通路，從而干預(yù)慢性咽炎。體內(nèi)實(shí)驗(yàn)結(jié)果表明，艾納香總黃酮能明顯改善大鼠咽部組織病理狀態(tài)；顯著降低大鼠血清中IL-6和PGE₂水平（ P＜0.05、0.01），升高IL-10水平（ P＜0.01）；顯著下調(diào)大鼠咽部組織TLR4、PI3KCA、磷脂酰肌醇-3激酶調(diào)節(jié)亞基1（ PI3KR1）、Akt1、核因子κB激酶亞基β抑制因子（ IKBKB）和NF-κB1 mRNA表達(dá)水平（ P＜0.01），TLR4、PI3K、Akt、核因子κB p65（ NF-κBp65）蛋白表達(dá)水平及其磷酸化水平（ P＜0.05、0.01）。結(jié)論 艾納香總黃酮可能通過調(diào)控TLR4/PI3K/Akt/NF-κB信號通路，減輕炎癥反應(yīng)，從而改善慢性咽炎。

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

國家中醫(yī)藥管理局高水平中醫(yī)藥重點(diǎn)學(xué)科建設(shè)項(xiàng)目（zyyzdxk-2023186）；2023年貴州“黔六味”道地藥材優(yōu)勢特色產(chǎn)業(yè)集群建設(shè)項(xiàng)目“貴州道地藥材種質(zhì)資源庫設(shè)備建設(shè)”；貴州中醫(yī)藥大學(xué)喀斯特藥用資源保護(hù)與創(chuàng)新利用科技創(chuàng)新人才團(tuán)隊(duì)項(xiàng)目