动物造模 z-bio 2021-10-15 644

　　Objective: To establish a method for isolating high-purity pulmonary microvascular endothelial cells, and observe the cells of wild-type mouse and receptor for advanced glycation end product (RAGE) knockout mice under lipopolysaccharide (LPS) stimulation. Changes in skeleton F-actin.

　　Method: Take the lungs of 6-8 weeks old wild-type C57 mice and RAGE knockout mice, digest them with type I collagenase, filter by nylon cells, and isolate the primary mouse lung microvascular endothelium by immunomagnetic bead method The cells (PMVEC) were identified by morphology and immunofluorescence, and were stimulated with LPS (1 mg/L) for different periods of time and observed the changes of cytoskeleton F-actin with a laser confocal microscope.

　　Results: Under the microscope, it can be seen that the primary cultured PMVECs were fusiform or polygonal, and the purity was identified by cell anti-factor-related antigen (ⅧF-Ag) staining. After LPS stimulation, the PMVEC skeleton of wild-type mice rearranged and formed stress fibers. However, the PMVEC skeleton of RAGE knockout mice is not obviously damaged.

　　Conclusion: High-purity mouse lung microvascular endothelial cells can be obtained by the immunomagnetic bead method, and RAGE is involved in the LPS-mediated destruction of mouse lung microvascular endothelial cytoskeleton.