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目的研究槲皮素对海水诱发性急性肺损伤(SW-ALI)的保护作用,并探索该作用与M1型巨噬细胞极化及其介导的炎症的相关性。方法雄性C57BL/6小鼠随机分成正常对照组、槲皮素处理组、SW-ALI模型组、SW-ALI联合槲皮素治疗组,每组15只。SW-ALI模型组小鼠采用10 g/L戊巴比妥钠腹腔注射麻醉,经口咽插管按4 mL/kg剂量匀速将海水注入肺中;SW-ALI联合槲皮素治疗组于海水注射前0.5 h给予槲皮素(200 mg/kg)腹腔注射;正常对照组、槲皮素处理组经腹腔注射等量PBS或槲皮素。各组小鼠在造模0.5、1、2、3、4 h后,于麻醉状态下行腹主动脉取血1 mL进行血气分析;3 d后,取1/3肺脏制备常规冰冻切片、HE染色观察肺组织病变;1/3肺脏称湿质量,80℃恒温箱烤48 h至干质量恒定,计算干/湿质量比(D/W);采用Western blot法检测肺组织中M1型巨噬细胞标志蛋白诱导型一氧化氮合酶(iNOS)的水平,实时定量PCR检测肺组织中iNOS及肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-4及IL-10的mRNA水平。结果槲皮素能明显减轻肺组织病理改变;降低二氧化碳分压(PaCO_2)并增加氧分压(PaO_2)和肺组织D/W;减少iNOS蛋白水平,下调TNF-α及IL-1β的水平且上调IL-4及IL-10的mRNA水平。结论槲皮素可通过抑制M1型巨噬细胞极化及其介导的免疫反应显著改善SW-ALI后气体交换功能、减轻肺组织病变。
Objective To investigate the protective effect of quercetin on seawater-induced acute lung injury (SW-ALI) and to explore its relationship with the polarization of M1 macrophages and the inflammation mediated by them. Methods Male C57BL / 6 mice were randomly divided into normal control group, quercetin treatment group, SW-ALI model group and SW-ALI combined with quercetin treatment group, 15 rats in each group. The SW-ALI model mice were anesthetized by intraperitoneal injection of pentobarbital at a dose of 10 g / L and saline was injected into the lungs at a constant dose of 4 mL / kg via oral orotracheal intubation. SW-ALI combined with quercetin treatment in seawater Quercetin (200 mg / kg) was intraperitoneally injected 0.5 h before injection. Normal control group and quercetin-treated group were injected intraperitoneally with equal volume of PBS or quercetin. Blood samples were obtained from the abdominal aorta of 1 ml for anesthetized mice after modeling for 0.5, 1, 2, 3 and 4 h respectively. Three days later, 1/3 of the lungs were used to prepare routine frozen sections and HE staining One third of the lungs were weighed and wet mass was weighed and then dried at 80 ° C for 48 h until the dry weight was constant. The dry / wet weight ratio (D / W) was calculated. Western blot was used to detect the expression of M1 macrophages The levels of iNOS, IL-1β, IL-4 and IL-6 in lung tissue were detected by real-time quantitative PCR. 10 mRNA level. Results Quercetin could significantly reduce the pathological changes of lung tissue, decrease the partial pressure of carbon dioxide (PaCO_2) and increase the partial pressure of oxygen (PaO_2) and the lung tissue D / W, reduce the level of iNOS protein and down-regulate the levels of TNF-α and IL- Upregulate mRNA levels of IL-4 and IL-10. Conclusion Quercetin can significantly improve the gas exchange function after SW-ALI and alleviate the pathological changes of lung tissue by inhibiting the polarization of M1 macrophage and its immune response.