Effect of astragaloside on adhesion of human umbilical vein endothelial cells and polymorphonuclear neutrophils and expression of nuclear factor kappa B under hypoxia/reoxygenation condition / 中国组织工程研究

ABSTRACT

AIM: To investigate the intervention effect of astragaloside on adhesion of polymorphonuclear neutrophils (PMN) and vascular endothelial cells and on expression of nuclear factor-?B (NF-?B) in human umbilical vein endothelial cells (hUVECs) injured by hypoxia/reoxygenation due to ischemia/reperfusion. METHODS: The experiment was performed at the Cell Bioengineering Laboratory of Institute of Cerebrovascular Diseases, Affiliated Hospital of Medical College, Qingdao University from September 2005 to May 2006. ①Neonatal umbilical cords were offered by Department of Gynecology and Obstetrics in Affiliated Hospital of Medical College of Qingdao University, with informed consent of puerperant and their families. The experiment was accorded with ethical standard of Helsinki declaration. ②The hUVECs were cultured to the future passage. After the third passage, hUVECs were randomly divided into three groups. Cells in a control group were cultured under normal conditions; Cells in a hypoxia/reoxygenation group were cultured under in closed container with hypoxia for 1 hour, and then under normal conditions for 1 hour; The hUVECs in a astragaloside group were pretreatment by different doses of astragaloside (20, 40, 80 mg/L). After 12 hours, hUVECs suffered from hypoxia/reoxygenation. ③The concentration of intercellular adhesion molecule (ICAM)-1 and content of malondialdehyde (MDA) in supernatant were detected by enzyme linked immunosorbent assay (ELISA) and thiobarbituric acid method. Expression of NF-?B of hUVECs was analyzed by immunohistochemistry. PMN adhesion to hUVECs was measured by rose Bengal staining. RESULTS: ①Compared to the control group, the content of MDA remarkably increased in hypoxia/reoxygenation group (P