Effects of rapamycin on puromycin aminonucleoside induced podocyte injury

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Objective To study the protective effect of mpamycin on puromycin aminonucleoside (PAN) induced podocyte injury and its possible mechanisms.Methods (1) The synchronized podocytes were divided into four groups:the control group (RPMI 1640 culture medium without serum) and the 20,50,100 mg/L PAN group (a final concentration of 20,50,100 mg/L PAN RPMI 1640 culture medium without serum).The cells were collected after 24 h before Western blot assay was performed to assess relative expression of Nephrin protein.(2) The synchronization podocytes were assigned to the model group and 10,100,1 000μg/L mpamycin group.All the cells were added to a final concentration of 50 mg/L PAN RPMI 1640 serum-free culture medium,incubated for 24 h.The model control group was added to dissolve the same amount of mpamycin solution,and the remaining three groups were added to a final concentration of 10,100,1 000μg/L rapamycin training.After 24 and 48 h,Western blot assay was used to test the relative expression of Nephrin protein.(3) The synchronization podocytes were assigned to thee normal control group (serum-free RPMI 1640 culture solution),model control group (the final concentration of 50 mg/L PAN + serum-free RPMI 1640 culture hquid) and mpamycin group (final concentration of 100 μg/L rapamycin + 50 mg/L PAN + serum free RPMI 1640 medium).The cells were cultured for 24 and 48 h,and their morphology was observed.Western blot was used to detect p-P70S6K relative expression.Results (1) Compared with the control group,Nephrin protein expression was significantly decreased in 50 or 100 mg/L PAN group (P ＜ 0.05).No significant difference in nephrin protein expression was found between 50 or 100 mg/L PAN group.(2) Compared with 10 or 1 000μg/L group,Nephrin protein expression was significantly increased in 100 μg/L mpamycin group (P ＜ 0.05).(3) Compared with the normal control group,Nephrin protein expression did not significantly change in the PAN or mpamycin group.Conclusions 50 mg/L PAN could damage the optimal concentration of PAN in vitro cell model.100 μg/L mpamycinon may protect podocytes from PAN induced injury,which may contribute to the inhibition of mTOR. Key words: Rapamycin; Podocytes; p-p70S6K; Nephrin