肿瘤坏死因子α中和抑制脂多糖诱导小鼠急性呼吸窘迫综合征肺组织细胞凋亡的机制探讨  

Lung Tissue Apoptosis Mechanism of LPS-induced ARDS in Mice via TNF-α Neutralization

作　　者：张安兵[1,2] 李理[1] 袁伟锋[1] 先俊 黄文杰[1] 

机构地区：[1]广州军区广州总医院呼吸内科,广东广州510010 [2]广州医科大学研究生学院,广东广州510182

出　　处：《中国呼吸与危重监护杂志》2014年第3期246-249,共4页Chinese Journal of Respiratory and Critical Care Medicine

基　　金：国家自然科学基金面上项目(编号:81070003;81370173);国家自然科学基金青年科学基金项目(编号:81200002);广东省社会发展领域科技计划项目(编号:20120318040)

摘　　要：目的探讨肿瘤坏死因子α(TNF-α)中和抑制脂多糖(LPS)诱导小鼠急性呼吸窘迫综合征(ARDS)肺组织细胞凋亡的机制。方法小鼠随机分为对照组、LPS组和TNF-α中和组。采用LPS(5 mg/kg)气道雾化造小鼠ARDS模型,TNF-α中和组在滴入LPS前24 h腹腔注射依那西普(0.4 mg/kg),滴入LPS 2 h后收集标本。PCR检测各组肺组织核转录因子κB(NF-κB)p65、Bax、Bcl-2的表达水平,Western blot检测NF-κB p65和Erk1/2及二者磷酸化、Bax、Bcl-2的蛋白水平;测量各组肺组织干湿重比;HE染色观察各组肺组织病理改变,采用肺损伤半定量评分评估肺组织损伤程度。结果 LPS组肺组织NF-κB及Erk1/2活化水平升高、Bcl-2与Bax比降低(P<0.05)。TNF-α中和能明显降低ARDS小鼠肺组织NF-κB活化水平,Bcl-2与Bax比值升高。TNF-α中和组小鼠肺组织湿干重比及肺损伤半定量评分较LPS组显著降低(P<0.05)。结论 TNF-α中和抑制脂多糖诱导小鼠ARDS肺组织损伤,其机制与抑制Erk1/2、NF-κB活化,上调Bcl-2/Bax比值,最终减少细胞凋亡密切相关。Objective To investigate the mechanism of lung tissue apoptosis in LPS-induced miceARDS via TNF-αneutralization.Methods Thirty-six mice were randomly divided into a control group, aLPS group, and TNF-αneutralization group. LPS（ 5 mg/kg） was intratracheally nebulized to induce ARDS inthe LPS group and the TNF-αneutralization group. Twenty-four hours before LPS treatment, etanercept （ 0.4mg/kg） was abdominal injected to the mice in the TNF-αneutralization group. Mice were sacrificed 2 hoursafter LPS treatment. PCR were used to detected the expression of NF-κB p65, Bax and Bcl-2 in lung tissue.Western blot were used to detected protein level of NF-κB p65, Erk1 /2 and their phosphorylation and Bax,Bcl-2. The lung dry-to-wet ratio was measured. The lung histological changes were evaluated by HE staining.Results Activation level of NF-κB p65 and Erk1 /2 was elevated, the ratio of Bcl-2 and Bax was decreased inthe LPS group（ P 〈 0. 05） . After TNF-αneutralization, the activation level of NF-κB p65 and Erk1 /2 werereduced, the ratio of Bcl-2 and Bax was increased （ P 〈 0. 05） . Compared with the LPS group, the lung dryto-wet ratio and lung injury semi-quantitative score were significantly decreased in the TNF-αneutralizationgroup （ P 〈0. 05） .Conclusion TNF-αneutralization can suppress lung injury in LPS-induced ARDSmiceby inhibiting activation of NF-κB p65 and Erk1/2, increasing the ratio of Bcl-2 and Bax ratio, and eventuallyreducing apoptosis.

关 键 词：急性呼吸窘迫综合征 脂多糖 肿瘤坏死因子Α 细胞凋亡 

分 类 号：R346[医药卫生—基础医学]