Rat Pulmonary Microvascular Endothelial Cell Senescence Mechanism

Aged pulmonary insufficiency is the possible common pathological process of many lung diseases , which can lead to multiple organ dysfunction syndrome(MODS),eventually death. It is urgent to study on molecular and cellular mechanisms of age-related diseases, including aged pulmonary insufficiency. Pulmonary microvascular endothelial cells( PMVECs )is the key point in pathogenesis of pulmonary insufficiency because of the special functions. We hope to clarify the celluar and molecular mechanisms of pulmonary insufficiency during ageing through research on the change rules by signal pathway in growth, survival and apoptosis . The subject not only will provide useful clues for the study of aged pulmonary insufficiency and age-related diseases but also has extremely important social and economic meaning..ObjectTo establish a stable and relialble model of PMVECs ageing induced by D-gal based on isolating and culturing PMVECs from rats lung successfully; Explore cellular and molecular mechanisms of PMVECs senescence on cell cycle, growth and survival ,apoptosis ,signal pathways.Methods1. Isolation, culture, purification and identification of PMVECs from rats lung. The purified PMVECs were obtained after isolated from peripheral lung tissues , digested with trypsin and collagenase and aggregated by serum stimulation. According to typical cobble morphology of endothelial cells, PMVECs were selected and transferred by trypsin/EDTA local digestion to amplify the number of cells. The following methods were used to identify PMVECs from rats lung.(l) von...