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作 者:陈丽华[1] 刘雪松[1] 刘飞[1] 金伯泉[1]
机构地区:[1]第四军医大学基础部免疫学教研室,西安710032
出 处:《生理学报》2003年第3期355-359,共5页Acta Physiologica Sinica
基 金:ThisworkwassupportedbytheNationalNaturalScienceFoundationofChina (No 30 0 30 130and 30 2 70 718)andKeyProgramofNationalNaturalScienceFoundationofChina (No 2 0 0 1CB5 10 0 0 4)
摘 要:为观察CD2 2 6单克隆抗体 (mAb)对培养人脐静脉内皮细胞 (HUVECs)胞质钙离子变化的影响 ,我们用Flu o 3作为钙指示剂 ,用激光共聚焦显微镜观测不同状态下CD2 2 6mAb作用后HUVECs胞质钙离子 [Ca2 +]i 的变化。结果发现 :( 1)用Hanks液平衡 ,CD2 2 6mAb作用后HUVECs [Ca2 +]i水平缓慢升高后回到原位 ;加入二抗 (羊抗鼠IgG)交联后 [Ca2 +]i水平有较大幅度的升高 ,随后回到原位 ,与此同时 ,细胞外液中 [Ca2 +]o水平有一定程度的下降 ;( 2 )用D Hanks液平衡 ,CD2 2 6mAb作用后HUVECs [Ca2 +]i水平无显著变化 ,加入二抗发生交联作用后 ,[Ca2 +]i水平有较大幅度的下降 ;( 3 )用EGTA预处理后 ,CD2 2 6mAb及其二抗交联对HUVECs [Ca2 +]i 变化无显著影响。以上结果提示 ,CD2 2 6mAb及其二抗交联可诱导不同状态的HUVECs胞质钙离子水平发生不同程度的变化 。In order to study the possible mechanism of CD226 monoclonal antibody (mAb)-mediated intracellular message transduction in human umbilical vein endothelial cells (HUVECs), the influence of CD226 mAb and its cross-linking by secondary antibody (ⅡAb) on the concentration changes in [Ca 2+] i in the HUVECs under different conditions were determined by confocal laser scanning microscopy. The main results are as follows. (1) When the culture medium was balanced by Hanks Buffer, [Ca 2+] i in HUVECs increased slowly after stimulation by CD226 mAb, whereas [Ca 2+] i increase was accompanied by [Ca 2+] o decrease after the mAb was cross-linked by goat anti-mouse IgG. Then [Ca 2+] i and [Ca 2+] o all returned to the normal level. (2) When the culture medium was balanced by D-Hanks buffer, [Ca 2+] i in HUVECs showed little variation when the cells were stimulated by CD226 mAb, but [Ca 2+] i decreased markedly after cross-linking. (3) When HUVECs were pretreated with EGTA, there was no variation in [Ca 2+] i of HUVECs after CD226 mAb stimulation alone or cross-linking of the mAb. Our results suggest that stimulation by CD226 mAb and cross-linking by goat anti-mouse IgG induce the variation of [Ca 2+] i in HUVECs under different conditions and the variation of [Ca 2+] i in HUVECs may play an important role in many physiological and pathological processes.
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