脂氧素A4改善肺微血管内皮细胞炎症损伤的体外实验研究  

Lipoxin A4 improves inflammatory lesions of human pulmonary microvascular endothelial cells in vitro

作　　者：陈云志[1] 连泽来 余华军[1] 孔鸿儒 戴胜杰 黄超豪 孙洪伟[1] CHEN Yun-zhi;LIAN Ze-lai;YU Hua-jun;KONG Hong-ru;DAI Sheng-jie;HUANG Chao-hao;SUN Hong-wei(Department of Hepatobiliary & Pancreatic Surgery,the First Affiliated Hospital,Wenzhou Medical University,Wenzhou,Zhejiang 325000,China;Phamacy Class 1 of Wenzhou Medical University,Wenzhou,Zhejiang 325035,China)

机构地区：[1]温州医科大学附属第一医院肝胆胰外科,浙江温州325000 [2]温州医科大学,药学一班浙江温州325035

出　　处：《肝胆胰外科杂志》2019年第4期233-237,共5页Journal of Hepatopancreatobiliary Surgery

基　　金：国家自然科学基金面上项目(81070372);温州市科技局项目(Y20100228)

摘　　要：目的观察脂氧素A4(lipoxin A4,LXA4)对肺微血管内皮细胞(pulmonary microvascular endothelial cells,PMVECs)炎症损伤的治疗作用。方法体外培养人PMVECs,第一阶段:分为空白对照组(完全培养基培养)和TNF-α组(分别以5、10、20 ng/mL共培养24 h),采用MTT法检测各组PMVECs存活率,用针头式滤器检测TNF-α损伤前后PMVECs的单层通透性。第二阶段:分为TNF-α组(20 ng/mL处理24 h)和LXA4干预组(1、10、100μg/L处理24 h),检测PMVECs存活率和单层通透性,RT-PCR检测PMVECs IL-1β、VCAM-1、NF-κB p65 mRNA的表达。结果 (1)5、10、20 ng/mL TNF-α组的PMVECs存活率分别是(76.39±9.3)%、(61.87±11.7)%、(49.54±12.6)%,与空白对照组比较,均显著降低(P <0.05),而经1、10、100μg/L三种浓度LXA4干预后,20 ng/mL TNF-α组的PMVECs存活率均得到提高,其中100μg/L LXA4干预后的PMVECs存活率为(82.32±8.7)%,有统计学差异(P <0.01);(2)与损伤前[(0.021±0.011)mL/min·cm^2·kPa]比较,TNF-α处理60、90min后PMVECs单层通透系数(Kf值)显著增高[(0.067±0.007)和(0.096±0.007)mL/min·cm^2·kPa,P <0.05)],而LXA4干预60、90 min时Kf值则显著降低[(0.039±0.008)和(0.058±0.011) mL/min·cm^2·kPa,P <0.05)];(3)与对照组比较,TNF-α组的IL-1β、VCAM-1、NF-κB p65 mRNA的表达水平均有显著升高(P <0.01或P <0.05),而LXA4干预组显著改善(P <0.01或P <0.05)。结论 TNF-α能引起PMVECs的急性炎症损伤,LXA4对TNF-α引起的PMVECs急性炎症损伤有改善作用。Objective To study the experimental healing effect of lipoxin A4 (LXA4) on inflammatory lesions of human pulmonary microvascular endothelial cells(PMVECs) which was induced by TNF-α in vitro. Methods The first stage: human pulmonary microvascular endothelial cells were randomly divided into two groups: sham group and TNF-α group(Final concentration of TNF-α was 5, 10, 20 ng/mL respectively;Co-cultivation period was 24 h). The survival rates were detected by MTT colormetric assay. The momolayer endothelial cells permeability of PMVECs was detected by filter needle. The second stage: PMVECs were randomly divided into two groups: TNF-α group (Final concentration of TNF-α was 20 ng/mL;Co-cultivation period was 24 h) and LXA4 group (Final concentration of LXA4 was 1, 10, 100 ug/l respectively;Co-cultivation period was 24 h). Similarly, the survival rates and the momolayer endothelial cells permeability of PMVECs were detected. The mRNA levels of IL-1β, VCAM-1 and NF-κB p65 were detected by RT-PCR assay. Results (1)The survival rates of PMVECs in 5, 10 and 20 ng/mL TNF-α group were (76.39±9.3)%,(61.87±11.7)% and (49.54±12.6)% respectively, compared with the sham group, decreased significantly (P<0.05). However, 1, 10 and 100 μg/L LXA4 could increase the survival rates in 20 ng/mL TNF-α group (P<0.05), especially 100 μg/L LXA4 increased significantly (82.32±8.7%, P<0.01).(2)Compared with the normal group (0.021±0.011 mL/min·cm2·kPa), the momolayer endothelial cells permeability of PMVECs (Kf value) in TNF-α group increased significantly after 60 and 90 min induced by TNF-α[(0.067±0.007) and (0.096±0.007) mL/min·cm2·kPa, P<0.05], however, Kf value decreased significantly when treated with LXA4 [(0.039±0.008) and (0.058±0.011) mL/min·cm2·kPa, P<0.05].(3)Compared with the sham group, the mRNA levels of IL-1β, VCAM-1 and NF-κB p65 of PMVECs in TNF-α group increased significantly (P<0.05), however, treatment with LXA4 can significantly decrease the mRNA levels (P<0.05). Conclusion TNF-α can in

关 键 词：脂氧素A4 肺微血管内皮细胞 炎症损伤 

分 类 号：R364[医药卫生—病理学]