动物造模 z-bio 2022-01-22 552

　　Objective: To establish a method for the isolation of high-purity pulmonary microvascular endothelial cells and observe the receptors of wild-type mouse cells and advanced glycation end products (RAGE) knock-out mice after lipopolysaccharide (LPS) stimulation.

　　Method: Collect the lungs of wild-type C57 and RAGE knockout mice aged 6-8 weeks, digest with type I collagenase, filter through nylon cells, and separate from the lungs of primary mice with immunomagnetic beads. .. Identify cells by morphology and immunofluorescence (PMVEC), stimulate different periods with LPS (1mg/L), and observe the changes of cytoskeleton F-actin under a laser confocal microscope.

　　Result: Under the microscope, it can be seen that the primary cultured PMVEC is spindle-shaped or polygonal, and it is highly purified by cell anti-factor VIII-related antigen (VIIIF-Ag) staining. After LPS stimulation, the PMVEC bones of RAGE knockout mice were not damaged, but wild-type mice rearranged to form stress fibers.

　　Conclusion: The immunomagnetic bead method can obtain high-purity mouse lung microvascular endothelial cells, and RAGE is involved in the LPS-mediated destruction of mouse lung microvascular endothelial cytoskeleton.