目的 探究厚朴酚抑制瞬时受体电位香草素4（TRPV4）活性缓解脂多糖（LPS）诱导小鼠急性肺水肿的作用。方法 SPF级昆明小鼠随机分为6组：对照组、模型组、盐酸多巴酚丁胺(DBT，阳性药，5 mg·kg^-1)组和厚朴酚低、中、高剂量(3.3、10.0、30.0 mg·kg^-1)组。除对照组外，其他各组建立LPS诱导的小鼠肺水肿模型，LPS滴注前30 min和滴注后8 h分别ip给药1次，检测肺湿/干质量比、病理损伤（HE染色）；试剂盒法检测肺泡灌洗液肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6、IL-1β及总蛋白水平；伊文思蓝法检测肺血管通透性；免疫荧光法检测紧密连接蛋白闭锁小带蛋白-1（ZO-1）表达。使用炔基修饰厚朴酚探针在小鼠肺组织切片和肺微血管内皮细胞（PMVEC）中验证厚朴酚与TRPV4共定位。体外培养PMVEC，分为对照组、模型组、GSK219(TRPV4抑制剂，10μmol·L^-1)组和厚朴酚低、中、高浓度(0.1、1.0、10.0μmol·L^-1)组，除对照组外使用LPS (10μmol·L^-1)处理24 h诱导损伤，造模的同时给药，活细胞钙成像检测胞内Ca²⁺浓度，鬼笔环肽染色观察细胞骨架，Western blotting检测ZO-1蛋白表达，Transwell实验检测跨内皮电阻（TEER）。结果 与对照组比较，厚朴酚(30 mg·kg^-1)显著降低肺湿/干质量比（P<0.05）、肺泡灌洗液总蛋白（P<0.05）及炎症因子水平（P<0.01），减轻肺组织病理损伤、血管渗漏及逆转ZO-1缺失。厚朴酚探针与TRPV4共定位良好。与对照组比较，厚朴酚(1μmol·L^-1)显著抑制LPS诱导的TRPV4介导Ca²⁺超载（P<0.001），减轻F-actin应力纤维收缩和细胞间连接断裂，上调ZO-1表达（P<0.01），恢复TEER（P<0.001）。结论 厚朴酚通过抑制TRPV4活性，调控肺微血管内皮细胞钙稳态和内皮屏障功能，有效缓解LPS诱导的急性肺水肿，为阐释中药厚朴“燥湿”功效提供科学依据。

Objective To investigate the effect and mechanism of magnolol in alleviating lipopolysaccharide（LPS）-induced acute pulmonary edema in mice through inhibition of transient receptor potential vanillin 4（TRPV4） activity. Methods SPF-grade Kunming mice were randomly divided into six groups: control group, model group, dobutamine hydrochloride(DBT, positive drug, 5 mg·kg^-1) group, and the magnolol low-, medium-, and high-dose(3.3, 10.0, 30.0 mg·kg^-1) groups. Except for control group, LPSinduced mouse pulmonary edema models were established in the other groups. Drugs were administered ip once 30 min before LPS injection and 8 h after LPS injection. The lung wet/dry weight ratio, pathological damage（HE staining）, and levels of tumor necrosis factor（TNF）-α, interleukin（IL）-6, IL-1β, and total protein in bronchoalveolar lavage fluid were detected. The lung vascular permeability was measured by Evans blue method, and the expression of tight junction protein zonula occludens-1（ZO-1） was detected by immunofluorescence. The co-localization of magnolol and TRPV4 was verified in mouse lung tissue sections and pulmonary microvascular endothelial cells（PMVEC） using an alkynyl-modified magnolol probe. PMVEC were cultured in vitro and divided into control group, model group, GSK219(TRPV4 inhibitor, 10 μmol·L^-1) group, and magnolol low-, medium-, and high-concentration(0.1, 1.0, 10.0 μmol·L^-1) groups. Except for control group, cells were treated with LPS(10 μmol·L^-1) for 24 h to induce injury, and drugs were administered simultaneously. Intracellular Ca²⁺ concentration was detected by live cell calcium imaging, F-actin stress fiber contraction and intercellular junction disruption were observed by phalloidin staining, and the expression of ZO-1 protein was detected by Western blotting. Transwell assay was used to detect transendothelial electrical resistance（TEER）. Results Compared with the control group, magnolol(30 mg·kg^-1) significantly reduced the lung wet/dry weight ratio（P＜0.05）, total protein in bronchoalveolar lavage fluid（P＜0.05）, and levels of inflammatory factors（P＜0.001）, alleviated lung tissue pathological damage, vascular leakage, and reversed ZO-1 deficiency. The magnolol probe co-localized well with TRPV4. Compared with the control group, magnolol(1 μmol·L^-1) significantly inhibited LPS-induced TRPV4-mediated Ca²⁺ overload（P＜0.001）, alleviated F-actin stress fiber contraction and intercellular junction disruption, upregulated ZO-1 expression（P＜0.01）, and restored TEER（P＜0.001）. Conclusion Magnolol alleviates LPS-induced acute pulmonary edema by inhibiting TRPV4 activity, thereby regulating calcium homeostasis and endothelial barrier function in PMVECs. This study provides scientific evidence elucidating the "drying-dampness" efficacy of the traditional Chinese medicine Magnolia officinalis.

R285.5

广西“带土移植”人才引育计划项目(桂科AA23026008); 天津市大学生创新科研计划项目(202410055330)