Effects Of Cd40-cd40 Ligand Interaction On Human Umbilical Vein Endothelial Cells

Objective 1. To take a suitable method for the culture and identification of human umbilical vein endothelial cells (HUVECs) and macrophages;2. The effects of CD40-CD40L interaction on HUVECs expression of notric oxide (NO) and intercellular adhesion molecule-1(ICAM-1) were observed by co-incubating HUVECs with the macrophages of human, the cell type than has been demonstrated to express CD40L in atherosclerotic plaque. By exploring the mechanism underlying plaque vulnerability, we attempted to provide new clues for the prevention and treatment of AS.Methods 1.Collagenase perfusion is adopted to isolate HUVEC, and trypsin is used for confluent culture. The staining of VIII is taken to ascertain the purity of HUVC. 2. Macrophages were erived from healthy human peripheral blood monocytes by density gradient centrifugation. Phagocytosis test and CD68 is taken to ascertain the macrophage.3 The third passage of HUVECs are used for experiment, and observate the effect of HUVEC by. co-incubating HUVECs with the macrophages.Results 1.Isolation of HUVEC with collagenase and the trypsin for confluent culture, and both satisfactory. The VIII stain confirmed the HUVEC; 2. The obtainned cells by density gradient centrifugation were highly pur-ified macrophages with favourable phagocytosis, acid phosphatase (ACP) and immu-nocytochemistry.3 the cell survival rate is lower in HUVECs with the macrophages than control group and anti-CD40L group at 24 hour and 48 hour (P<0.05),and lower obviously in 48 hour than 24 hour (P<0.05).4. ICAM-1 is increase obviously in HUVECs with the macrophages than control group and anti-CD40L group at 24 hour and 48 hour (P<0.05),and increase obviously in48 hour than 24 hour (P<0.05).5.NO is reduced obviously in HUVECs with the macrophages than control group and anti-CD40L group at 24 hour and 48 hour (P<0.05),and reduced obviously in 48 hour than 24 hour (P<0.05)Conclusions 1.Isolation of HUVEC with collagenase and the trypsin for confluent culture, and both satisfactory. The VIII stain confirmed the HUVEC; 2. The obtainned cells by density gradient centrifugation were highly pur- ified macrophages with favourable phagocytosis, acid phosphatase (ACP) and immu-nocytochemistry.3 CD40-CD40L significantly enhanced HUVECs increased the ICAM-1, and reduced NO.4.CD40-CD40L signaling may ruduces to the activation of endothelial cells.