摘要
目的探讨白头翁皂苷B4对脂多糖(Lipopolysaccharide,LPS)诱导的急性肺损伤(AcuteLungInjury,ALI)的保护作用及其潜在的作用机制。方法采用RAW264.7细胞进行细胞实验,用B4预先给药后,LPS刺激(100ng·mL^-1)24h,然后测定培养液中一氧化氮(NitricOxide,NO)、肿瘤坏死因子-α(TumorNecrosisFactor,TNF-α)、白介素-6(Interleukin-6,IL-6)、白介素1β(Interleukin-1β,IL-1β)的浓度。将BALB/c小鼠随机分为正常对照组,模型组,地塞米松组(5mg·kg^-1),B4高、中、低(2、4、8mg·kg^-1)剂量组。给药组小鼠腹腔注射不同剂量的药物,正常对照组和模型组给予生理盐水;地塞米松组、B4组及模型组鼻腔内滴入LPS建立急性肺损伤模型,24h后收集支气管肺泡灌洗液(BronchoalveolarLavageFluid,BALF),全自动血液分析仪检测BALF中白细胞、中性粒细胞的数量,酶联免疫吸附法测定BALF中TNF-α,IL-1β和IL-6的水平,苏木素—伊红(HE)染色观察肺组织病理形态的改变。蛋白免疫印迹法(WesternBlot)测定肺组织中NF-κB蛋白表达水平。结果在细胞实验中,白头翁皂苷B4能够显著降低NO、TNF-α、IL-1β和IL-6的高表达(P<0.05,P<0.01)。体内实验中,与正常对照组相比,模型组小鼠BALF的中性粒细胞数量、TNF-α、IL-1β和IL-6的含量,肺组织损伤,以及NF-κB的蛋白表达均有显著上升(P<0.05,P<0.01);与模型组相比,白头翁皂苷B4低剂量组可以显著减少中性粒细胞的数量,降低TNF-α,IL-6的含量,改善肺组织损伤并下调NF-κB的蛋白表达(P<0.05,P<0.01)。结论白头翁皂苷B4能够有效改善LPS诱导的肺部损伤,其作用机制可能与下调NF-κB蛋白,降低相关炎性因子的释放有关。
Objective To investigate the protective effect of Pulsatilla saponin B4 on acute lung injury(AU)induced by lipopolysaccharide(LPS)and its potential mechanism.Methods RAW 264.7 cells were used for cell experiments.After pre-administration with B4,LPS was stimulated(100 ng·mL^-1)for 24 hours,and then the concentrations of Nitric Oxide(NO),Tumor Necrosis Factor(TNF-α),Interleukin-6(IL-6),Interleukin-1β(IL-1β)in the culture medium were determined.BALB/c mice were randomly divided into normal group,model group,dexamethasone group,B4 high,medium,low(2,4,8 mg·kg^-1)dose groups.Mice were intraperitoneally injected with different closes of B4.Normal group and model group rats were given normal saline.LPS was instilled into the nasal cavity to establish acute lung injuiy model.Bronchoalveolar lavage fluid(BALF)was collected 24 hours later and analyzed by automatic blood analyzer to measuer the numbers of leukocytes and neutrophils.The levels of TNF-α,IL-1βand IL-6 in BALF were determined by enzyme-linked immunosorbent assay(ELISA).Pathological morphology of lung tissues was examined by hematoxylin-eosin(HE)staining.The expression level of NF-κB protein in lung tissue was determined by Western Blot.Results In the cell experiment,Pulsatilla saponin B4 significantly decreased the expression of NO,TNF-α,IL-10 and IL-6(P<0.05,P<0.01).In vivo,compared with the normal group,the number of neutrophils,TNF-α,IL-1βand IL-6,lung injury and NF-κB protein expression in BALF of the model group were significantly increased(P<0.05,P<0.01).Compared with the model group,the low dose group of Pulsatilla saponin B4 can significantly reduce the number of neutrophils,reduce the contents of TNF-α,IL-6,improve lung tissue damage and down-regulate the protein expression of NF-κB(P<0.05,P<0.01).Conclusion Pulsatilla saponin B4 can effectively improve LPS-induced lung injury,and its mechanism may be related to down-regulation of NF-κB protein and reducing the release of related inflammatory factors.
作者
周朦静
陈兰英
胡宏辉
罗颖颖
方聪
张妮
ZHOU Mending;CHEN Lanying;HU Honghui;LUO Yingying;FANG Cong;ZHANG Ni(National Engineering Research Center for Solid Pharmaceutical Manufacturing Technology,Jiangxi University of Traditional Chinese Medicine,Nanchang 330006 Jiangxi,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2019年第6期664-670,共7页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
江西省重点研发计划(20181ACG70014)
国家自然基金项目(81860720)
