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
发布时间：2018-10-21 19:21
【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(P0.05);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(P0.05),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(P0.05),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。
[Abstract]:Aim to observe the adhesion characteristics of rat pulmonary artery endothelial cells (rat pulmonary arterial endothelial cell,RPAEC) and rat aortic endothelial cells (rat aortic endothelial cell,RAEC) to monocytes (U937) under hypoxia. To investigate the difference of adhesion between endothelial cells and monocytes in pulmonary circulation and systemic circulation under hypoxia. Methods three-air incubator was used to simulate hypoxia environment (1%O_2) and normoxic control (21%O_2) cells were cultured in normal incubator. The adhesion of endothelial cells to U937 cells was studied by single treatment, according to the type of endothelial cells and the different conditions of hypoxia treatment. The experiment was divided into six groups: normoxic RPAEC normoxic U937 group (CPE CU), hypoxia RPAEC U937 (HPE CU), hypoxia RPAEC hypoxia U937 group (HPE HU), RAEC normoxic U937 group (CAE CU), anoxic RAEC normoxic U937 (HAE CU) and hypoxia RAEC hypoxia U937 group (HAE HU) (nun3). Flow cytometry was used to detect the proportion of Di I cells after cell adhesion test, fluorescence enzyme labeling instrument was used to detect the fluorescence intensity of Di I in cell lysate after adhesion test, cell adhesion was observed and counted by inverted microscope, and the adhesion rate of U937 cells was calculated. Adhesion rate of endothelial cells and adhesion of endothelial cells. Results flow cytometry showed that the rate of Di I cells in, HPE CU group was significantly higher than that in CPE CU group, and the difference was statistically significant (P0.05 between); HPE CU and HPE HU group, there was no significant difference). Fluorescence enzyme labeling assay showed that the fluorescence value in the cell lysate in, HPE CU group was significantly higher than that in CPE CU group (P0.05), but there was no significant difference between RAEC and U937 adhesion test groups (P0.05), but there was no significant difference between RAEC and U937 adhesion test groups. The adhesion of RPAEC to U937 cells was observed by inverted fluorescence microscope. The results showed that the adhesion rate and adhesion force of U937 cells in, HPE CU group were significantly higher than those in CPE CU group (P0.05), although the adhesion rate of U937 cells in, HPE CU group was higher than that in CPE CU group (P0.05), and the adhesion rate of U937 cells to U937 cells in, HPE CU group was significantly higher than that in CPE CU group. But there was no significant difference between. HPE CU group and HPE HU group. Conclusion hypoxia specifically stimulates the adhesion of RPAEC from pulmonary circulation to U937, and whether U937 is stimulated by hypoxia has no effect on the adhesion of U937 to RPAEC.
【作者单位】： 重庆医科大学干细胞与组织工程研究室;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;
【基金】：国家自然科学基金(30971198,81270108)
【分类号】：R563
[Abstract]:Aim to observe the adhesion characteristics of rat pulmonary artery endothelial cells (rat pulmonary arterial endothelial cell,RPAEC) and rat aortic endothelial cells (rat aortic endothelial cell,RAEC) to monocytes (U937) under hypoxia. To investigate the difference of adhesion between endothelial cells and monocytes in pulmonary circulation and systemic circulation under hypoxia. Methods three-air incubator was used to simulate hypoxia environment (1%O_2) and normoxic control (21%O_2) cells were cultured in normal incubator. The adhesion of endothelial cells to U937 cells was studied by single treatment, according to the type of endothelial cells and the different conditions of hypoxia treatment. The experiment was divided into six groups: normoxic RPAEC normoxic U937 group (CPE CU), hypoxia RPAEC U937 (HPE CU), hypoxia RPAEC hypoxia U937 group (HPE HU), RAEC normoxic U937 group (CAE CU), anoxic RAEC normoxic U937 (HAE CU) and hypoxia RAEC hypoxia U937 group (HAE HU) (nun3). Flow cytometry was used to detect the proportion of Di I cells after cell adhesion test, fluorescence enzyme labeling instrument was used to detect the fluorescence intensity of Di I in cell lysate after adhesion test, cell adhesion was observed and counted by inverted microscope, and the adhesion rate of U937 cells was calculated. Adhesion rate of endothelial cells and adhesion of endothelial cells. Results flow cytometry showed that the rate of Di I cells in, HPE CU group was significantly higher than that in CPE CU group, and the difference was statistically significant (P0.05 between); HPE CU and HPE HU group, there was no significant difference). Fluorescence enzyme labeling assay showed that the fluorescence value in the cell lysate in, HPE CU group was significantly higher than that in CPE CU group (P0.05), but there was no significant difference between RAEC and U937 adhesion test groups (P0.05), but there was no significant difference between RAEC and U937 adhesion test groups. The adhesion of RPAEC to U937 cells was observed by inverted fluorescence microscope. The results showed that the adhesion rate and adhesion force of U937 cells in, HPE CU group were significantly higher than those in CPE CU group (P0.05), although the adhesion rate of U937 cells in, HPE CU group was higher than that in CPE CU group (P0.05), and the adhesion rate of U937 cells to U937 cells in, HPE CU group was significantly higher than that in CPE CU group. But there was no significant difference between. HPE CU group and HPE HU group. Conclusion hypoxia specifically stimulates the adhesion of RPAEC from pulmonary circulation to U937, and whether U937 is stimulated by hypoxia has no effect on the adhesion of U937 to RPAEC.
【作者单位】： 重庆医科大学干细胞与组织工程研究室;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;
【基金】：国家自然科学基金(30971198,81270108)
【分类号】：R563
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