急性肺损伤炎症环境中TNF-α介导肺通透性增加的机制研究  

Mechanism of TNF-α increasing pulmonary permeability in the inflammatory environment of acute lung injury

作　　者：李方治[1] 侯春阳[2] 孙佳英[1] 

机构地区：[1]中国医科大学附属第四医院呼吸内科,辽宁沈阳110032 [2]沈阳市第六人民医院ICU,辽宁沈阳110006

出　　处：《临床和实验医学杂志》2017年第19期1914-1918,共5页Journal of Clinical and Experimental Medicine

摘　　要：目的探讨急性肺损伤(ALI)时,炎症环境中肿瘤坏死因子(TNF-α)介导肺通透性增加的机制。方法30只SPF级健康Balb/c小鼠分为6组,对照组、0 h实验组、1 h实验组、2 h实验组、4 h实验组、8 h实验组,每组5只。实验组采用腹腔注射脂多糖(LPS)法,根据体重用要求的剂量于不同时间点进行注射,对照组注入同等剂量的生理盐水。LPS诱导急性肺损伤模型后,眼眶取血制备血清,游离肺组织碾磨制备上清,ELISA检测肺组织上清液中TNF-α的含量;LPS诱导急性肺损伤模型后,尾静脉注射伊文思蓝,在620 nm波长下检测不同时间点伊文思蓝溶液的吸收值;Western Blotting检测急性肺损伤小鼠模型中肺组织的低氧诱导因子-1α(HIF-1α)和血管扩张刺激磷蛋白(VASP)的蛋白表达水平改变。结果 LPS能成功诱导急性肺损伤模型,4 h实验组和8 h实验组的ELISA结果显示,实验组肺组织上清液中的TNF-α的含量明显升高;LPS造模后,尾静脉注射伊文思蓝,在620 nm波长下检测伊文思蓝溶液的吸收值,各组1 h开始升高,4 h实验组较0 h实验组明显升高,且差异有显著性(P<0.01);在小鼠的急性肺损伤模型中,实验组小鼠肺组织中HIF-1α的蛋白表达在4 h和8 h明显增加,同时VASP的蛋白表达在4 h和8 h明显降低。结论急性肺损伤症过程中,TNF-α可诱导HIF-1α的活化,进而引起VASP的表达下调,肺泡-毛细血管屏障损害,随着时间的增长,情况有所减退。Objective To investigate the effects and mechanisms of TNF-α increasing the tight junction permeability in lung tissue associated with acute lung inflammation. Methods Healthy Balb/c mice were divided into 6 groups： control group,0 h experimental group,1 h experimental group,2 h experimental group,4 h experimental group,8 h experimental group,5 rats in each group. The experimental group was injected with abdominal cavity,injected with the required dose according to the weight,and the control group was injected with the same dose normal saline. TNF-α in the serum and lung tissue were deby ELISA in LPS-induced acute lung injury mice model; Evans blue was used to detect the permeability of pulmonary capillary. Western Blotting was used to analyze HIF-1α and VASP protien expression level during the development of acute lung injury. Results In the LPS-induced model of ALI,the experiment showed that the concentration of TNF-α increased at 1 h and at4 h it reached peak by Elisa. Evans Blue was injected in tail vein,and then Evans Blue＇ absorption value was detected at 620 nm wavelength which reached its peak at 4 h. After LPS-inducing acute lung injury,HIF-1αprotein expression in the mice lung tissue increased significantly at 4 h and 8 h,while VASP protein expression decreased. Conclusion During acute pulmonary inflammation process,VASP is down regulated through TNF-α inducing HIF-1α activation,which plays an important role in the impairment of alveolar-capillary barrier.

关 键 词：小鼠 急性肺损伤 VASP TNF-Α 细胞通透性 

分 类 号：R563.8[医药卫生—呼吸系统]