摘要
目的分析腺苷A_(2B)受体(A_(2B) R)激活对脓毒症诱导的急性肺损伤(ALI)的影响并探讨其在肺微血管内皮炎症损伤中的调控作用。方法(1)将24只SD大鼠随机分为假手术组(sham组)、ALI模型组(ALI组)、A_(2B) R激动剂BAY60-6583干预组(ALI+BAY组)和A_(2B) R抑制剂PSB1115干预组(ALI+PSB组),每组6只。制模后24 h处死大鼠取肺组织,光镜下行肺损伤评分(Smith评分),计算肺湿/干质量比值(W/D)。Evans Blue染色法测定肺水清除率(AFC)。酶联免疫吸附试验(ELISA)测定肺组织炎症因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)的水平。(2)采用TNF-α诱导人肺微血管内皮细胞(HPMECs)损伤模型,设立对照组、TNF-α组、BAY组、PSB组、BAY+TNF-α组和PSB+TNF-α组,各组细胞培养24 h后采用异硫氰酸荧光素(FITC)-白蛋白(albumin)法检测HPMECs单层通透性,Werstern blot法检测IL-1β、细胞间黏附分子-1(ICAM-1)、血管内皮钙黏连蛋白(VE-cadherin)、促血管生成素(ANGPT)的表达水平。结果与sham组比较,ALI组的Smith评分、肺W/D及肺组织TNF-α、IL-6、IL-1β水平均显著升高,AFC显著降低;与ALI组比较,ALI+BAY组的Smith评分、肺W/D及肺组织TNF-α、IL-6、IL-1β水平显著降低,AFC显著升高;而ALI+PSB组结果相反。TNF-α诱导HPMECs损伤模型中,与对照组比较,TNF-α组IL-1β、ICAM-1表达水平及HPMECs单层通透性升高,VE-cadherin、ANGPT表达水平降低。与TNF-α组比较,BAY+TNF-α组IL-1β、ICAM-1表达水平及HPMECs单层通透性降低,VE-cadherin、ANGPT表达水平升高,而PSB1115预处理可逆转上述现象。上述各组之间差异有统计学意义(均P<0.05)。结论腺苷A_(2B) R激活可减轻脓毒症诱导的ALI,并通过减轻肺微血管内皮炎症、降低通透性、促进血管生成等途径起保护作用。
Objective To analyse the effect of the activation of adenosine A_(2B) receptor(A_(2B) R)on sepsis-induced acute lung injury(ALI)and to explore its regulatory role in pulmonary microvascular endothelial inflammation injury.Methods(1)A total of 24 Sprague-Dawley(SD)rats were randomly divided into sham operation group(sham group),ALI model group(ALI group),A_(2B) R agonist BAY60-6583 intervention group(ALI+BAY group),and A_(2B) R inhibitor PSB1115 intervention group(ALI+PSB group),with 6 rats in each group.The rats were sacrificed 24 hours after modeling,and their lung tissues were collected.Lung injury score(Smith score)was obtained under light microscopy,and lung wet/dry mass ratio(W/D)was calculated.Alveolar fluid clearance(AFC)rate was detected by Evans Blue staining.The levels of inflammatory factors in lung tissues including tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β)were detected by enzyme-linked immunosorbent assay(ELISA).(2)TNF-α-induced human pulmonary microvascular endothelial cells(HPMECs)injury model was used.The control group,TNF-αgroup,BAY group,PSB group,BAY+TNF-αgroup and PSB+TNF-αgroup were set up.After 24 hours of cell culture,fluorescein isothiocyanate(FITC)-albumin assay was used to detect the monolayer permeability of HPMECs in each group,and the levels of IL-1β,intercellular adhesion molecule-1(ICAM-1),vascular endothelial-cadherin(VE-cadherin)and angiopoietins(ANGPT)expressions were detected by Werstern blot.Results Compared with the sham group,the ALI group showed significantly increased Smith score,lung W/D,and levels of TNF-α,IL-6 and IL-1βin lung tissues,and showed significantly decreased AFC.Compared with the ALI group,the ALI+BAY group showed significantly decreased Smith score,lung W/D,and levels of TNF-α,IL-6 and IL-1βin lung tissues,and showed significantly increased AFC.However,the ALI+PSB group showed the opposite results.In the TNF-α-induced HPMECs injury model,compared with the control group,the TNF-αgroup showed the increased levels
作者
王慧霞
安友仲
WANG Huixia;AN Youzhong(Department of Critical Care Medicine,Peking University People′s Hospital,Beijing 100044,China)
出处
《中国临床新医学》
2024年第2期138-144,共7页
CHINESE JOURNAL OF NEW CLINICAL MEDICINE
基金
北京市自然科学基金项目(编号:7212124)。
关键词
腺苷A_(2B)受体
脓毒症
急性肺损伤
人肺微血管内皮细胞
炎症
Adenosine A_(2B)receptor(A_(2B)R)
Sepsis
Acute lung injury(ALI)
Human pulmonary microvascular endothelial cells(HPMECs)
Inflammation
