發(fā)布時(shí)間：2018-10-21 19:21

【摘要】：目的觀察缺氧條件下,大鼠肺動(dòng)脈內(nèi)皮細(xì)胞(rat pulmonary arterial endothelial cell,RPAEC)、大鼠主動(dòng)脈內(nèi)皮細(xì)胞(rat aortic endothelial cell,RAEC)與單核細(xì)胞(U937)黏附特性的變化。探討缺氧條件下,肺循環(huán)和體循環(huán)血管內(nèi)皮細(xì)胞與單核細(xì)胞黏附特性的差異。方法利用三氣培養(yǎng)箱模擬缺氧環(huán)境(1%O_2),常氧對(duì)照(21%O_2)細(xì)胞培養(yǎng)于普通培養(yǎng)箱。內(nèi)皮細(xì)胞與U937細(xì)胞為單獨(dú)處理后進(jìn)行黏附實(shí)驗(yàn),根據(jù)內(nèi)皮細(xì)胞類型以及缺氧處理?xiàng)l件的不同,實(shí)驗(yàn)分為6組:常氧RPAEC+常氧U937組(CPE+CU),缺氧RPAEC+常氧U937(HPE+CU),缺氧RPAEC+缺氧U937組(HPE+HU),常氧RAEC+常氧U937組(CAE+CU),缺氧RAEC+常氧U937(HAE+CU)和缺氧RAEC+缺氧U937組(HAE+HU)(n=3)。采用流式細(xì)胞術(shù)檢測(cè)細(xì)胞黏附實(shí)驗(yàn)后Di I+細(xì)胞比例;采用熒光酶標(biāo)儀檢測(cè)黏附試驗(yàn)后細(xì)胞裂解液中Di I熒光強(qiáng)度值;倒置顯微鏡觀察細(xì)胞黏附并計(jì)數(shù),計(jì)算U937細(xì)胞黏附率、內(nèi)皮細(xì)胞黏附率和內(nèi)皮細(xì)胞黏附力。結(jié)果流式檢測(cè)結(jié)果顯示,HPE+CU組Di I+細(xì)胞率顯著高于CPE+CU組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05);HPE+CU與HPE+HU組間,差異無顯著意義。熒光酶標(biāo)儀檢測(cè)結(jié)果顯示,HPE+CU組細(xì)胞裂解液中熒光值顯著高于CPE+CU組(P0.05),與HPE+HU組無顯著差異。RAEC與U937黏附實(shí)驗(yàn)各組間差異均無顯著意義。倒置熒光顯微鏡觀察RPAEC與U937細(xì)胞的黏附,結(jié)果顯示,HPE+CU組與CPE+CU組相比,U937細(xì)胞黏附率、內(nèi)皮細(xì)胞黏附力均顯著增加,差異具有統(tǒng)計(jì)學(xué)意義(P0.05),雖內(nèi)皮細(xì)胞黏附率相應(yīng)增高,但無統(tǒng)計(jì)學(xué)差異。HPE+CU組與HPE+HU組相比,各指標(biāo)差異無顯著性意義。結(jié)論缺氧特異性刺激肺循環(huán)來源的RPAEC與U937黏附;U937是否經(jīng)缺氧刺激對(duì)其與RPAEC的黏附?jīng)]有影響。
[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.
【作者單位】： 重慶醫(yī)科大學(xué)干細(xì)胞與組織工程研究室;第三軍醫(yī)大學(xué)高原軍事醫(yī)學(xué)系高原生理學(xué)與高原生物學(xué)教研室;
【基金】：國家自然科學(xué)基金(30971198,81270108)
【分類號(hào)】：R563

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