氢溴酸山莨菪碱对感染性休克大鼠急性肺损伤的保护作用

Protective effect of anisodamine hydrobromide on acute lung injury in rats with septic shock via the NF-κB/NLRP3 pathway

目的探讨氢溴酸山莨菪碱(Ani HBr)对感染性休克大鼠急性肺损伤的保护作用及其机制。方法将30只SD大鼠随机分为假手术(Sham)组、模型(LPS)组和Ani HBr治疗(LPS+Ani HBr)组,每组10只。利用气道滴注脂多糖(LPS)构建感染性休克大鼠急性肺损伤模型。建模1 h后,LPS+Ani HBr组大鼠腹腔注射3.6 mg/kg Ani HBr。Sham组和LPS组大鼠分别给予等量0.9%氯化钠溶液。药物干预6 h后,大鼠麻醉处死,取肺组织测定肺系数、肺组织湿/干重比(W/D)和肺组织含水量;同时,苏木精-伊红(HE)染色,观察肺组织的病理结构变化。收集支气管肺泡灌洗液(BALF),用酶联免疫吸附试验(ELISA)检测BALF中促炎细胞因子(IL-1β,IL-6和TNF-α)的表达水平。Western blotting检测肺组织中Bcl2,Bax和Caspase-3等凋亡相关蛋白表达以及NF-κB/NLRP3信号通路的活性。结果与LPS组比较,Ani HBr能够降低感染性休克大鼠的肺系数、肺组织湿/干重比(W/D)和肺组织含水量;改善受损大鼠肺泡结构紊乱,减少炎性细胞浸润;显著降低BALF中促炎细胞因子IL-1β、IL-6及TNF-α含量;减少肺组织中细胞凋亡水平,同时下调NF-κB p65和NLRP3蛋白表达水平。结论Ani HBr能够减轻感染性休克大鼠急性肺损伤,其分子机制与调控NF-κB/NLRP3信号通路相关。

Objective To investigate the protective effect of anisodamine hydrobromide(AniHBr)on acute lung injury in rats with septic shock and the underlying mechanism.Methods A total of 30 Sprague-Dawley(SD)rats were randomly divided into sham group,lipopolysaccharide(LPS)group and LSP+AniHBr group,with 10 rats in each group.The septic shock model in rats with acute lung injury was created by tracheal drip of LPS.After 1 hour of modeling,rats in LPS+AniHBr group were intraperitoneally injected with AniHBr 3.6mg/kg,and those in sham group and LPS group were similarly injected with 0.9%NaCl solution.After 6 hours of drug intervention,rats were sacrificed after anesthesia.Rat lung tissues were collected for measuring the lung index,wet/dry weight ratio(W/D)and water content.Pathological changes in the lung were observed by hematoxylin&eosin(H&E)staining.Bronchoalveolar lavage fluid(BALF)was collected to measure the contents of interleukin(IL)-1β,IL-6 and tumor necrosis factor-α(TNF-α)by Enzyme-linked immunosorbent assay(ELISA).Western blot was used to detect the protein expressions of apoptosis proteins like Bcl2,Bax and Caspase-3,as well as targeted proteins involved in the NF-κB/NLRP3 pathway in lung tissue.Results Compared with those of LPS group,rats in LPS+AniHBr group presented significantly lower lung index,W/D,water content in lung tissues,contents of IL-1β,IL-6 and TNF-αin BALF,and protein levels of apoptosis proteins,NF-κB p65 and NLRP3 in lung tissues,alleviated alveolar structure disorder,and reduced inflammatory infiltration.Conclusion AniHBr might alleviate LPS-induced acute lung injury in rats with septic shock by regulating the NF-κB/NLRP3 pathway.