3-甲基嘌呤对脂多糖诱导小鼠急性肺损伤的保护作用及其机制研究  

Protective Effects of 3-Methyladenine against Lipopolysaccharide-induced Acute Lung Injury in Mice and the Mechanism Study

作　　者：朱贲贲 马晓鹂[3] 吴科锋 陈华玲[1,2] 

机构地区：[1]广东天然药物开发研究实验室,广东湛江524023 [2]广东医学院药理教研室,广东湛江524023 [3]广东医学院附属医院,广东湛江524001

出　　处：《中国药房》2016年第4期464-468,共5页China Pharmacy

基　　金：广东省自然科学基金自由申请重点博士启动项目(No.S2012010008145)

摘　　要：目的:研究自噬抑制剂3-甲基嘌呤(3-MA)对脂多糖(LPS)诱导小鼠急性肺损伤的保护作用及其机制。方法:取小鼠随机分为正常对照组、模型(LPS 15 mg/kg)组、药物对照(3-MA 20 mg/kg)组和低、高剂量治疗(LPS 15 mg/kg+3-MA 20、40 mg/kg)组,每组10只。除正常对照组和药物对照组外,其余各组小鼠ip LPS复制急性肺损伤模型,药物对照组和低、高剂量治疗组小鼠分别于建模前1 h ip相应剂量的3-MA。建模6 h后分别测量各组小鼠的肺湿/干质量比(W/D),HE染色观察肺组织病理变化;Western blot法检测肺组织肿瘤坏死因子α(TNF-α)、NF-κB p65、LC3BⅡ/Ⅰ、激活型半胱氨酸氨基蛋白酶3(Cleaved-caspase-3蛋白的表达水平。结果:与正常对照组比较,模型组小鼠的W/D值和TNF-α、NF-κB p65、LC3BⅡ/Ⅰ、Cleaved-caspase-3蛋白表达均增强(P<0.01);与模型组比较,低剂量治疗组小鼠的W/D值和TNF-α、NF-κB p65、LC3BⅡ/Ⅰ、Cleaved-caspase-3蛋白表达均减弱(P<0.05),高剂量治疗组小鼠仅LC3BⅡ/Ⅰ蛋白表达减弱(P<0.01)。结论:在LPS诱导的急性肺损伤小鼠模型中,过度自噬可能通过激活NF-κB通路参与炎症反应并诱导细胞凋亡;3-MA适度抑制自噬可减轻炎症反应并起到保护肺组织的作用。OBJECTIVE：To study the protective effects of autophagy inhibitor 3-Methyladenine（3-MA）against lipopolysaccharide（LPS）-induced acute lung injury in mice and its mechanism. METHODS：Mice were randomly divided into normal control group,model group（LPS 15 mg/kg）,drug control group（3-MA 20 mg/kg）,low-dose and high-dose groups（LPS 15 mg/kg ＋3-MA 20,40 mg/kg）,with 10 mice in each group. Except for normal control group and drug control group,other groups were given LPS intraperitoneally to induce acute lung injury model,and drug control group and low-dose and high-dose groups were given equivalent dose of 3-MA intraperitoneally 1 h before modeling. 6 h after modeling,lung wet/drug mass ratio（W/D） was determined respectively,and pathology change of lung tissue was observed by HE staining. TNF-α,NF-κ B p65,LC3 B Ⅱ/Ⅰ and Cleaved-caspase-3 protein expression were detected by Western blot. RESULTS：Compared with normal control group,W/D,TNF-α,NF-κ B p65,LC3 B Ⅱ/Ⅰ and Cleaved-caspase-3 protein expression increased in model group（P〈0.01）. Compared with model group,W/D,the expression of TNF-α,NF-κB p65,LC3BⅡ/Ⅰ and Cleaved-caspase-3 protein decreased in low-dose group（P〈0.05）,white just only LC3BⅡ/Ⅰ protein decreased high-dose group（P〈0.01）. CONCLUSIONS：In LPS-induced acute lung injury model in mice,the excessive autophagy could activate the NF-κB pathway and involve the inflammatory responses and induce lung cells apoptosis. The moderate autophagy inhibition by 3-MA can ameliorate inflammatory response and protect lung tissue.

关 键 词：自噬 3-甲基嘌呤 急性肺损伤 脂多糖 小鼠 

分 类 号：R965[医药卫生—药理学]