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Effects of CD40-CD40 Ligand Interaction on Human Umbilical Vein Endothelial Cells

Author: YanFang
Tutor: SunZuo
School: Shihezi University
Course: Internal Medicine
Keywords: Human umbilical vein endothelial cells Macrophages Nitric oxide Intercellular adhesion molecule-1
CLC: R543.5
Type: Master's thesis
Year: 2008
Downloads: 4
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Abstract


The purpose of: 1. Explore the the human umbilical vein endothelial cells (HUVEC) and the macrophages cells culture with the identification of method; 2. Adopt the human umbilical vein endothelial cells with the able to expression that the a CD40L of the macrophages cells of common incubated for, as well as antibody retardation assay with method, to explore CD40-CD40L interaction on the the endothelial cells to the release of NO and the the impact of of ICAM-1, trying to explore the the artery porridge kind of hardening the pathological physiological mechanism on the the the level of of in the cellular and molecular, for the the the clinical prevention and treatment to provide new ideas. Method: 1. Adopt that the the type Ⅰ collagenase perfusion method to collect HUVEC, trypsin digestion the passaged, Ⅷ factor Free. Phytophthora group oriented the dyeing to identification of the the endothelial cells; 2. Through the the to the the density gradient centrifugation method from the healthy adults peripheral blood to obtain the single-core cells, and derived for the macrophages, to phagocytic the the the test and CD68 of immunohistochemistry the dyeing identification of; 3. Fetch the the third generation of HUVEC, observation of are of common incubated for with the macrophages., as well as adopt the the of the function changes in of of the HUVECs after the the anti-CD40L to monoclonal antibody blocking the CD40L the role of: (1) the inverted microscope to cell morphology was observed; (2) MTT method was used to detection the the the cell survival rate; (3) dual-antibody the sandwich enzyme-linked immunosorbent adsorption assay (ELISA) determination of ICAM-1; (4) nitric acid reductase method. The measured plasma NO. Results: 1. Of collagenase perfusion digestion and Law the cells were collected, trypsin digestion the passaged availability of the satisfied with the of the of HUVEC, Ⅷ factor staining confirmed that to cultured cells for the the endothelial cells; 2. Density gradient centrifugation method be able to to to obtain in the the from the human peripheral blood the high purity and activity of of the macrophages, phagocytosis test and the CD68 dyeing confirmed to be macrophages; 3. with the the to the macrophages co-incubated for the the group cell morphology occurrence of change the, 24 hours obvious with the the cell survival rate of the 48 hours lower than the that for the control group with the anti-CD40L group (P lt-; co-incubated for on 0.05) and the with the macrophages cells group survival rate the 48 hours cells lower than the 24 hours (P lt; 0.05). 4. With the the macrophages co-incubated for group in the 24-hour, The The the ICAM-1 the concentration of of the 48 hours significantly higher than that of the control group with the anti-of CD40-L group (P lt; 0.05), and the 48 hours the concentration of higher than the 24 hours (P lt; 0.05 ). 5. With the the macrophages co-incubated for group in the 24-hour, the the concentration of NO in of the 48 hours was significantly lower than the control group with the anti-of CD40-L group (P lt; 0.05), and the 48 hours the concentration of lower than the 24--hour (P lt; 0.05). On 1 collagenase perfusion of the Conclusion: digestion method cells were collected, trypsin digestion passaged obtained satisfactory HUVEC Ⅷ factor staining confirmed cultured cells for Endothelial Cells; 2 Density gradient centrifugation method be able to Get in the the from human peripheral blood high-purity and activity-of macrophages, The the phagocytosis test and the CD68 staining confirmed that for the the the macrophages cells; the the 3CD40-CD40L signaling pathways be able to to promote to HUVECs the release of ICAM-1, but but play an inhibitory effect on NO. 4 blocking that the the the signal system of the CD40-CD401 is could possibly delay the for the the of the the occurrence of the development of of of the vascular lesions.

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CLC: > Medicine, health > Internal Medicine > Heart, blood vessels ( circulatory ) disease > Vascular disease > Artery disease
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