Establishment of a podocyte cell injury model induced by puromycin aminonucleoside / 北京大学学报(医学版)

ABSTRACT

Objective:To establish a podocyte cell injury model induced by puromycin aminonucleoside(PAN),an in vitro model for studying the role of podocytes,especial the slit diaphragm molecules in proteinuria at the cellular and molecular levels.Methods:MPC5 were treated for 24 and 48 hours by 15,45 and 75 mg/L PAN,respectively.The podocyte molecular behavior during podocyte injury was evaluatedthe apoptotic podocyte cells were revealed with FITC-Annexin V and Propidium Iodide(PI) assay and the proliferative podocyte cells detected with MTT assay after PAN treatment.The distribution of Nephrin and Podocin was revealed with indirect-immnofluorescent staining under confocal microscope.The distribution of F-actin was revealed with direct-immnofluorescent staining under microscope.Results:The percentage of apoptotic podocyte cells was increased in a dose-and time-dependent manner after PAN treatment.In PAN-treated group,the apoptosis was obviously increased at hour 48,the PAN-45 treated group was 33.48%?14.55% and PAN-75 treated group 38.01%?12.13% vs the control group 6.38%?0.50%(P