| 摘要: |
| ［摘要］ 目的 分析腺苷A2B受体（A2BR）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）、A2BR激动剂BAY60-6583干预组（ALI+BAY组）和A2BR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A2BR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。 |
| 关键词: 腺苷A2B受体 脓毒症 急性肺损伤 人肺微血管内皮细胞 炎症 |
| DOI：10.3969/j.issn.1674-3806.2024.02.03 |
| 分类号:R 631 |
| 基金项目:北京市自然科学基金项目（编号：7212124） |
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| Activation of adenosine A2B receptor attenuates sepsis-induced acute lung injury and pulmonary microvascular endothelial inflammation injury |
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WANG Huixia, AN Youzhong
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Department of Critical Care Medicine, Peking University People′s Hospital, Beijing 100044, China
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| Abstract: |
| ［Abstract］ Objective To analyse the effect of the activation of adenosine A2B receptor(A2BR) 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), A2BR agonist BAY60-6583 intervention group(ALI+BAY group), and A2BR 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-α group, BAY group, PSB group, BAY+TNF-α group and PSB+TNF-α group 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-1β in 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-1β in 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-α group showed the increased levels of IL-1β and ICAM-1 expressions, and the increased monolayer permeability of HPMECs, and the decreased levels of VE-cadherin and ANGPT expressions. Compared with the TNF-α group, the BAY+TNF-α group showed the decreased levels of IL-1β and ICAM-1 expressions and the decreased monolayer permeability of HPMECs, and the increased levels of VE-cadherin and ANGPT expressions. However, pretreatment with PSB1115 could reverse the above phenomenon. The differences among the above groups were statistically significant(P<0.05). Conclusion Activation of A2BR can attenuate sepsis-induced ALI and exert protective effects by reducing pulmonary microvascular endothelial inflammation, reducing permeability and promoting angiogenesis pathways. |
| Key words: Adenosine A2B receptor(A2BR) Sepsis Acute lung injury(ALI) Human pulmonary microvascular endothelial cells(HPMECs) Inflammation |
