目的 探究草麻黃Ephedra sinica中一種鞣質(zhì)化合物2β, 3β-epoxyfavan-5, 7, 4′-triol-(4α→8)-epiafzelechin（Ete）對(duì)卵清白蛋白（ovalbumin，OVA）致敏哮喘小鼠的干預(yù)作用，為豐富麻黃抗哮喘的研究以及拓展麻黃臨床應(yīng)用提供實(shí)驗(yàn)依據(jù)。方法SPF級(jí)雌性BALB/c小鼠適應(yīng)性喂養(yǎng)1周，隨機(jī)分為對(duì)照組、模型組、陽(yáng)性對(duì)照藥孟魯司特鈉（1.314 mg/kg）組和Ete低、高劑量（10、20 mg/kg）組，采用ip OVA與Al₂(OH)₃混懸液聯(lián)合OVA霧化激發(fā)的方法建立哮喘小鼠模型，持續(xù)給藥1周。通過(guò)檢測(cè)小鼠咳嗽和喘息次數(shù)、氣道高反應(yīng)性、血清免疫球蛋白E（immunoglobulin E，IgE）以及小鼠肺部組織病理變化初步評(píng)價(jià)Ete抗OVA致敏哮喘的藥效；多指標(biāo)流式聯(lián)合分析系統(tǒng)檢測(cè)與哮喘密切相關(guān)的因子水平；流式細(xì)胞術(shù)檢測(cè)小鼠肺組織活性氧、凋亡和鈣離子水平以及小鼠機(jī)體免疫細(xì)胞（樹突狀細(xì)胞、嗜酸性粒細(xì)胞和肥大細(xì)胞）水平；ELISA檢測(cè)小鼠血清白三烯C4（leukotrienes C4，LTC4）、組胺（histamine，HIS）、β-肥大細(xì)胞類胰蛋白酶（β-mast cell tryptase，MCT-β）水平；Western blotting檢測(cè)肺組織中轉(zhuǎn)化生長(zhǎng)因子-β1（transforming growth factor-β1，TGF-β1）、Smad3、MUC5AC和表皮生長(zhǎng)因子受體（epidermal growth factor receptor，EGFR）蛋白表達(dá)水平。結(jié)果 與模型組比較，Ete顯著抑制哮喘小鼠氣道高反應(yīng)性，降低血清IgE水平（P＜0.01），減少咳、喘次數(shù)，延長(zhǎng)咳、喘潛伏時(shí)間（P＜0.05、0.01），降低血清中IL-13、TNF-α、CCL11水平（P＜0.05、0.01），升高IL-10水平（P＜0.05），降低肺組織中活性氧、凋亡、樹突狀細(xì)胞和嗜酸性粒細(xì)胞水平（P＜0.05、0.01），降低肥大細(xì)胞及其脫顆粒過(guò)程關(guān)鍵介質(zhì)LTC4、HIS、MCT-β和MCP-1的水平（P＜0.05、0.01），降低黏液分泌相關(guān)蛋白TGF-β1、Smad3、EGFR和MUC5AC的水平（P＜0.05、0.01）。結(jié)論 草麻黃中一種鞣質(zhì)化合物Ete通過(guò)抑制肥大細(xì)胞活化和黏液蛋白分泌來(lái)改善OVA誘導(dǎo)的小鼠過(guò)敏性哮喘。

Objective To explore the intervention effect of a tannin compound 2β, 3β-epoxyfavan-5, 7, 4′-triol-(4α→8)-epiafzelechin (Ete), extracted from Ephedra sinica on ovalbumin (OVA)-sensitized asthmatic mice, providing experimental evidence to enrich the research on anti-asthmatic properties of Mahuang (Ephedrae Herba) and expand its clinical applications. Methods SPF female BALB/c mice were acclimatized for one week and randomly divided into control group, model group, positive drug montelukast (1.314 mg/kg) group and Ete low-, high dose (10, 20 mg/kg) groups. An asthma mouse model was established by intraperitoneal injection of OVA and Al₂(OH)₃ suspension combined with OVA atomization excitation, with continued administration for one week. The efficacy of Ete against OVA-sensitized asthma was evaluated by detecting cough and wheezing numbers, airway hyperresponsiveness, serum immunoglobulin E (IgE) and pathological changes of lung tissue in mice. Multi index flow based joint analysis system was used to detect the levels of factors closely related to asthma. Flow cytometry was used to detect the levels of reactive oxygen species (ROS), apoptosis and Ca²⁺ in lung tissue of mice and the levels of immune cells (dendritic cells, eosinophils and mast cells) in mice. The levels of serum leukotrienes C4 (LTC4), histamine (HIS) and β-mast cell tryptase (MCT-β) in mice were detected by ELISA. The protein expression levels of transforming growth factor-β1 (TGF-β1), Smad3, MUC5AC and epidermal growth factor receptor (EGFR) in lung tissue were detected by Western blotting. Results Compared with model group, Ete significantly inhibited airway hyperreactivity in asthmatic mice, reduced serum IgE level (P < 0.01), decreased the frequency of coughing and wheezing, and prolonged the latency for coughing and wheezing (P < 0.05, 0.01), lowered serum IL-13, TNF-α and CCL11 levels (P < 0.05, 0.01), while increased IL-10 level (P < 0.05), reduced levels of ROS, apoptosis, dendritic cells and eosinophils in lung tissue (P < 0.05, 0.01), decreased the levels of mast cells and key mediators involved in their degranulation process (LTC4, HIS, MCT-β and MCP-1) (P < 0.05, 0.01), reduced the levels of mucin-secreting proteins TGF-β1, Smad3, EGFR and MUC5AC (P < 0.05, 0.01). Conclusion A tannin compound Ete from E. sinica improves OVA-induced allergic asthma in mice by inhibiting mast cell activation and mucin secretion.

R285.5

國(guó)家重點(diǎn)研發(fā)計(jì)劃—中醫(yī)藥現(xiàn)代化研究專項(xiàng)（2019YFC1708802）；國(guó)家自然科學(xué)基金資助項(xiàng)目（32200322）；河南省高層次人才特殊支持計(jì)劃“中原千人計(jì)劃”—中原領(lǐng)軍人才（ZYQR201810080）；河南省科技研發(fā)計(jì)劃聯(lián)合基金（242301420082）