醋酸烯诺孕酮对脂多糖诱导的急性肺损伤的调节作用

The effect of nestorone on pulmonary inflammation in lipopolysaccharides-induced acute lung injury

目的:通过体内、体外实验探讨醋酸烯诺孕酮(nestorone,NES)对脂多糖(lipopolysaccharides,LPS)诱导的急性肺损伤(acute lung injury,ALI)模型的调节作用.方法:通过体外培养A549细胞,以MTT法检测细胞存活率并以酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)测定炎性细胞因子分泌.建立ALI小鼠模型并以NES预干预,以肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中总细胞计数、细胞分类计数并通过H&E检测各组肺部炎症反应程度,同时分析肺干湿比和BALF总蛋白含量.最后检测NES对ALI小鼠生存状态的影响.结果:NES无细胞毒性(10-4~100μmol·L-1)且NES(1和10μmol·L-1)显著抑制LPS诱导的A549细胞促炎因子的分泌.体内研究发现NES的预处理可有效缓解小鼠肺部急性炎症损伤,表现为降低BALF总细胞数、中性粒细胞数、巨噬细胞数和淋巴细胞数,减少蛋白渗出,降低炎性因子[白介素-6(IL-6)、趋化因子配体1(CXCL-1)和肿瘤坏死因子-α(TNF-α)]的分泌且延长小鼠生存期和生存率.结论:NES可显著抑制LPS诱导的A549细胞和ALI小鼠肺部的炎症反应,延长ALI小鼠生存期和生存率.

Objective:To investigate the regulatory effect of nestorone(NES)on lipopolysaccharides(LPS)-induced acute lung injury(ALI)model by in vivo and in vitro experiments.Methods:A549 cells were cultured to analyze the cell survival rate by MTT and the pro-inflammatory cytokines by enzyme-linked immunosorbent assay(ELISA).A mouse model of LPS-induced ALI was established and pre-treated with NES.The acute inflammatory response within lung was evaluated by total cell counts,differential cell counts in bronchoalveolar lavage fluid(BALF),the degree of lung inflammation in each group by H&E,the lung ratio of wet and dry,and total proteins in BALF.Then the protective effect of NES on survival status was examined by LPS-induced ALI model.Results:In A549 cells,there was no cytotoxicity in the experimental concentrations(10-4~100μmol·L-1),and NES(1 and 10μmol·L-1)significantly inhibited LPS-induced inflammatory cytokines production.In the LPS-induced ALI mouse model,NES pre-treatment significantly attenuated the acute inflammatory response and damage in lung,including reducing the number of total cells,neutrophils,macrophages,and lymphocytes in BALF,reducing proteins exudation,and decreasing the expression of inflammatory cytokines(IL-6,CXCL-1 and TNF-α).The survival time and survival rate were increased by NES.Conclusion:NES has potent anti-inflammatory activity in LPS-induced A549 cells and ALI mouse model,prolong the survival period and increase the survival rate of ALI mice.