[Establishment of a podocyte cell injury model induced by puromycin aminonucleoside].
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 evaluated: the 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-immunofluorescent staining under confocal microscope. The distribution of F-actin was revealed with direct-immunofluorescent 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<0.01).


CONCLUSIONS
We set up an in vitro podocyte injury model treated with PAN for the first time. This reliable cell model is a good basis for further studies on podocyte injury.
Liu
Shu Fang
SF
Department of Pediatrics, Peking University First Hospital, Beijing, China.
Ding
Jie
J
Fan
Qing Feng
QF
Zhang
Han
H
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D051379:Mice; D023421:Models, Animal; D050199:Podocytes / Q000473:pathology*; D011692:Puromycin Aminonucleoside
D011692:Puromycin Aminonucleoside