目的 研究柚皮苷對(duì)脂多糖（LPS）誘導(dǎo)的小鼠急性肺損傷模型肺微血管內(nèi)皮通透性及氣道炎癥的改善作用。方法 將66只C57BL/6雄性小鼠隨機(jī)分為對(duì)照組、模型組、地塞米松組及柚皮苷40、80、120 mg/kg組，每組11只。對(duì)照組小鼠正常飼養(yǎng)；模型組和給藥組小鼠麻醉后給予LPS滴鼻，每只小鼠滴鼻50 μL。柚皮苷40、80、120 mg/kg組和地塞米松5 mg/kg組在LPS滴鼻前分別ig相應(yīng)劑量的柚皮苷和地塞米松，連續(xù)給藥5 d，滴鼻后繼續(xù)ig相應(yīng)劑量的柚皮苷和地塞米松，連續(xù)給藥2 d。對(duì)照組和模型組同時(shí)間ig 0.2 mL生理鹽水。蘇木精–伊紅（HE）染色觀察肺實(shí)質(zhì)病理變化；血常規(guī)分析儀檢測(cè)血液及肺泡灌洗液淋巴細(xì)胞、中性粒細(xì)胞、血細(xì)胞、血小板數(shù)量；ELISA檢測(cè)肺泡灌洗液白細(xì)胞介素（IL）-1β、IL-6、腫瘤壞死因子-α（TNF-α）含量和血清內(nèi)皮素-1（ET-1）水平；肺組織伊文思藍(lán)成像分析肺泡–血管通透性，qPCR檢測(cè)緊密連接蛋白緊密連接蛋白-1（ZO-1）、密封蛋白（OCLN）、血管內(nèi)皮鈣黏素（VE-cadherin）、β-連環(huán)蛋白（β-catenin）和水通道蛋白AQP1、AQP5的mRNA表達(dá)水平；化學(xué)分析法檢測(cè)血清丙二醛（MDA）、超氧化物歧化酶（SOD）、一氧化氮（NO）水平。結(jié)果 與模型組相比，柚皮苷120 mg/kg組能夠顯著改善LPS誘導(dǎo)的急性肺損傷小鼠肺水腫及肺組織炎癥浸潤(rùn)的病理狀態(tài)，減少肺泡內(nèi)炎癥細(xì)胞數(shù)量、炎癥因子和總蛋白含量，以及肺組織伊文思藍(lán)滲入量；增強(qiáng)緊密連接蛋白和水通道蛋白表達(dá)；減緩血液炎癥細(xì)胞流失，降低血管氧化應(yīng)激水平（P＜0.05）。結(jié)論 柚皮苷能夠減輕LPS誘導(dǎo)的氣道炎癥，并可能通過(guò)降低血管氧化應(yīng)激水平減少對(duì)緊密鏈接蛋白和水通道蛋白的損害作用從而改善肺內(nèi)皮高通透性。

Objective To study the effect of naringin on pulmonary microvascular endothelial permeability and airway inflammation in mice model of acute lung injury induced by lipopolysaccharide (LPS). Methods Sixty-six male C57BL/6 mice were randomly divided into control group, model group, dexamethasone group, and naringin 40, 80, 120 mg/kg groups, with 11 mice in each group. Control group mice were fed normally. Mice in the model group and the administration group were given LPS nasal drops after anesthesia, each mice was given 50 μL nasal drops. Naringin 40, 80, and 120 mg/kg groups and dexamethasone 5 mg/kg groups were intragastrically given naringin and dexamethasone at corresponding doses before LPS nasal drops for 5 days, respectively, and intragastrically given naringin and dexamethasone at corresponding doses after nasal drops for 2 days. Control group and model group were given 0.2 mL normal saline at the same time. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of lung parenchyma. The number of lymphocytes, neutrophils, blood cells and platelets in blood and alveolar lavage fluid were detected by blood routine analyzer. The contents of interleukin-1β, IL-6, tumor necrosis factor-α (TNF-α) and serum endothelin-1 (ET-1) in alveolar lavage fluid were determined by ELISA. Evens blue imaging of lung tissue to analyze alveolar-vascular permeability, qPCR was used to detect the mRNA expression levels of tight junction protein-1 (ZO-1), sealing protein (OCLN), vascular endothelial cadherin (VE-cadherin), β-catenin (β-catenin) and aquaporins AQP1 and AQP5. Serum levels of malondialdehyde (MDA), superoxide dismutase (SOD) and nitric oxide (NO) were detected by chemical analysis. Results Compared with model group, naringin 120 mg/kg group could significantly improve the pathological state of pulmonary edema and inflammatory infiltration of lung tissue in mice with LPS-induced acute lung injury, reduce the number of inflammatory cells in alveolar, the contents of inflammatory factors and total protein, and the infiltration amount of Evans blue in lung tissue. Enhance the expression of tight junction protein and aquaporin; decrease the loss of blood inflammatory cells and decrease the level of vascular oxidative stress (P < 0.05). Conclusion Naringin can reduce LPS-induced airway inflammation, and may improve pulmonary endothelial hyperpermeability by reducing vascular oxidative stress levels and reducing damaging effects on tightlink protein and aquaporin.

R285

浙江省中醫(yī)藥科技計(jì)劃項(xiàng)目(2022ZB225)