缺氧对肺循环和体循环血管内皮细胞与单核细胞黏附特性的影响  

Hypoxia specifically promotes the adhesion of pulmonary vascular endothelial cells to monocytes

作　　者：陈婷[1] 谭小玲[2] 杨诚忠 李满满[1] 杨戎[1] 

机构地区：[1]重庆医科大学干细胞与组织工程研究室,400016 [2]第三军医大学高原军事医学系高原生理学与高原生物学教研室,重庆400038

出　　处：《免疫学杂志》2017年第4期288-293,共6页Immunological Journal

基　　金：国家自然科学基金(30971198;81270108)

摘　　要：目的观察缺氧条件下,大鼠肺动脉内皮细胞(rat pulmonary arterial endothelial cell,RPAEC)、大鼠主动脉内皮细胞(rat aortic endothelial cell,RAEC)与单核细胞(U937)黏附特性的变化。探讨缺氧条件下,肺循环和体循环血管内皮细胞与单核细胞黏附特性的差异。方法利用三气培养箱模拟缺氧环境(1%O_2),常氧对照(21%O_2)细胞培养于普通培养箱。内皮细胞与U937细胞为单独处理后进行黏附实验,根据内皮细胞类型以及缺氧处理条件的不同,实验分为6组:常氧RPAEC+常氧U937组(CPE+CU),缺氧RPAEC+常氧U937(HPE+CU),缺氧RPAEC+缺氧U937组(HPE+HU),常氧RAEC+常氧U937组(CAE+CU),缺氧RAEC+常氧U937(HAE+CU)和缺氧RAEC+缺氧U937组(HAE+HU)(n=3)。采用流式细胞术检测细胞黏附实验后Di I+细胞比例;采用荧光酶标仪检测黏附试验后细胞裂解液中Di I荧光强度值;倒置显微镜观察细胞黏附并计数,计算U937细胞黏附率、内皮细胞黏附率和内皮细胞黏附力。结果流式检测结果显示,HPE+CU组Di I+细胞率显著高于CPE+CU组,差异具有统计学意义(P<0.05);HPE+CU与HPE+HU组间,差异无显著意义。荧光酶标仪检测结果显示,HPE+CU组细胞裂解液中荧光值显著高于CPE+CU组(P<0.05),与HPE+HU组无显著差异。RAEC与U937黏附实验各组间差异均无显著意义。倒置荧光显微镜观察RPAEC与U937细胞的黏附,结果显示,HPE+CU组与CPE+CU组相比,U937细胞黏附率、内皮细胞黏附力均显著增加,差异具有统计学意义(P<0.05),虽内皮细胞黏附率相应增高,但无统计学差异。HPE+CU组与HPE+HU组相比,各指标差异无显著性意义。结论缺氧特异性刺激肺循环来源的RPAEC与U937黏附;U937是否经缺氧刺激对其与RPAEC的黏附没有影响。To study the adhesion property to monocytes of vascular endothelial cells from pulmonarycirculation and systemic circulation under hypoxia condition, we analyzed the adhesion of rat pulmonary arteryendothelial cells（RPAEC） and rat aortic endothelial cells（RAEC） to monocyte U937. The experimental rats weredivided into six groups： normoxic RPAEC＋normoxic U937（CPE＋CU）, hypoxic RPAEC＋normoxic U937（HPE＋CU）,hypoxic RPAEC ＋ hypoxic U937（HPE ＋ HU）, normoxic RAEC ＋ normoxic U937（CAE ＋ CU）, hypoxic RAEC ＋normoxic U937（HAE ＋ CU） and hypoxic RAEC ＋ hypoxic U937（HAE ＋ HU）. Flow cytometry, fluorescencequantitative analysis and fluorescence microscopy were used to detect the monocyte adhesion state. Flow cytometryanalysis showed that the percent of Di I＋cell in HPE＋CU group was much higher than that of CPE＋CU group（P〈0.05）, but there had no obvious difference between HPE＋HU and HPE＋CU groups on the percent of Di I＋cell.RAEC adhesion to U937 cell between hypoxic and normoxic conditions. Fluorescence density of celllysates from the group of HPE ＋ CU was much higherthan that from CPE＋CU group（P〈0.05）, while no difference of the density was existed between HPE＋HU and HPE＋CU groups. Similarly, there was no difference in fluorescence density in the cell lysates from RAEC and U937 celladhesion groups. We further detected the cell adhesion of U937 with hypoxic RPAEC by fluorescence microscope.The results showed that monocyte adhesion rate and endothelial cell adhesion ability were significantly increased inHPE＋CU group compared with CPE＋CU（P〈0.05）. However, no difference was detected between HPE＋HU andHPE＋CU groups. In conclusion, hypoxia could specifically promote the adhesion of RPAEC cells to U937 cells, nomatter U937 treated with or without hypoxia.

关 键 词：肺动脉内皮细胞 主动脉内皮细胞 单核细胞 细胞黏附 缺氧 

分 类 号：R563[医药卫生—呼吸系统]