金线莲提取物对脂多糖诱导的小鼠急性肺损伤的保护作用及机制研究  

Protective effect and mechanism of Anoectochilus roxburghii extract on acute lung injury induced by LPS in mice

作　　者：朱亮 包晓君 王群星[3] 徐菲拉[1] ZHU Liang;BAO Xiaojun;WANG Qunxing;XU Feila(Centers of Traditional Medicine,Jinhua Municipal Central Hospital,Jinhua 321000,China;不详)

机构地区：[1]金华市中心医院传统医学中心,321000 [2]浙江金华天宁堂生物科技有限公司 [3]金华市中心医院药学部,321000

出　　处：《浙江医学》2024年第4期341-346,I0003,共7页Zhejiang Medical Journal

基　　金：金华市科技计划项目(2021-4-281);浙江省医学会临床科研基金项目(2020ZYC-A126)。

摘　　要：目的 观察金线莲提取物对脂多糖(LPS)诱导的急性肺损伤(ALI)模型小鼠的疗效并探讨其作用机制。方法 60只健康ICR小鼠随机分为正常组、模型组、地塞米松组和金线莲低、中、高剂量组。地塞米松组给予5.0 mg/kg地塞米松溶液腹腔注射,金线莲低、中、高剂量组分别给予1.3、2.6、5.2 g/kg的金线莲提取物灌胃,对照组和模型组给予0.9%氯化钠溶液灌胃,连续给药2 d。第3天对照组给予0.9%氯化钠溶液气管滴注,其余各组均给予5.0 mg/kg LPS构建ALI模型。造模后各组再给药1次。12 h后颈椎脱臼处死小鼠,获取肺组织及肺泡灌洗液。HE染色观察肺组织病理学变化,测定肺湿干比,ELISA法测定肺泡灌洗液中IL-6、单核细胞趋化蛋白-1(MCP-1)、IL-12/p70、IFN-γ、TNF-α和IL-10的水平,免疫组化法检测肺组织中髓过氧化物酶(MPO)、磷酸化细胞外信号调节激酶(p-ERK)、人核因子κB抑制蛋白α(IκBα)和磷酸化人核因子κB抑制蛋白α(p-IκBα)的蛋白表达水平,Western blot法检测肺组织中细胞外信号调节激酶(ERK)、p-ERK、c-Jun氨基末端激酶(JNK)、磷酸化c-Jun氨基末端激酶(p-JNK)、p38、磷酸化p38(p-p38)、腺苷酸-活化蛋白激酶α(AMPKα)、磷酸化腺苷酸-活化蛋白激酶α(p-AMPKα)、MPO蛋白相对表达量。结果 与对照组相比,模型组小鼠肺组织出现炎性病理变化,肺泡灌洗液中IL-6、MCP-1、IL-12/p70、IFN-γ、TNF-α和IL-10水平均显著升高(均P<0.01)。与模型组相比,金线莲高剂量组肺组织湿干比明显降低(P<0.01),肺泡灌洗液中IL-6、MCP-1、IL-12/p70、IFN-γ、TNF-α和IL-10水平均显著降低(均P<0.05),肺组织MPO、p-ERK、pIκBα蛋白表达水平均显著降低(均P<0.05),p-ERK、p-JNK、p-p38、p-AMPKα、MPO蛋白相对表达量均显著降低(均P<0.05)。结论 一定剂量金线莲提取物可有效保护LPS所致ALI,其作用机制可能与调节丝裂原活化蛋白激酶信号通路、减轻炎症反应有关。Objective To investigate the effects of Jinxianlian(JXL,Anoectochilus roxburghii) extract on lipopolysaccharide(LPS)-induced acute lung injury in mice and explore its mechanism.Methods Sixty healthy ICR mice were randomly divided into control group(CON),model group(LPS),dexamethasone(DEX) group,low-dose JXL extract(JXL-L)group,medium-dose JXL extract(JXL-M) group,high-dose JXL extract(JXL-H) group.DEX solution(5.0 mg/kg) was given by intraperitoneal injection in DEX group,different doses of JXL extract solution(1.3,2.6,5.2 mg/kg) were given by gavage in the JXL-L,JXL-M,and JXL-H groups,while the blank group and model group were given normal saline by gavage for 2 d.On the D3,mice in the normal group were instilled with normal saline through the nose,and the other groups were instilled with LPS(5.0 mg/kg) to induce acute lung injury(ALI).Twelve hours after the last administration,mice were sacrificed,and lung tissue and bronchoalceolar lavage fluid(BALF) samples were collected.The lung wit/dry weight ratio(W/D) was measured,and HE staining was used to observe the pathological changes of lung tissue.The levels of IL-6,monocyte chemoattractant protein-1(MCP-1),IL-12/p70,IFN-γ,TNF-α and IL-10 in BALF were measured by ELISA.The relative expression of myeloperoxidase(MPO),phosphorylated activated extracellular signal-regulated kinase(p-ERK),nuclear factor kappa-B(IκBα) and phosphorylated IκBα(p-IκBα) in lung tissue were measured by immunohistochemistry(IHC).The expression of related proteins ERK,p-ERK,c-Jun N-terminal kinase(JNK),phosphorylated JNK(p-JNK),p38,phosphorylated p38(p-p38),adenosine monophosphate actived protein kinase α(AMPKα),phosphorylated AMPKα(p-AMPKα),MPO in lung tissue was detected by Western blot.Results Compared with the control group,the model group displayed inflammatory pathological changes in lung tissue,and the levels of IL-6,MCP-1,IL-12/p70,IFN-γ,TNF-α and IL-10 in BALF were increased(all P<0.01).Compared with the model group,the pathological changes of the JXL-H group were atte

关 键 词：金线莲 急性肺损伤 丝裂原活化蛋白激酶信号通路 炎症因子 

分 类 号：R285.5[医药卫生—中药学]