Role and its mechanism of micro RNA-130b in puromycin aminonucleoside-induced podocyte injury

Objective To investigate the role and its mechanism of miR-130b in podocyte injury induced by puromycin aminonucleoside(PAN).Methods The immortalized mouse podocytes MPC5 were treated by 25, 50, 100 μg/mL PAN, then real-time quantitative polymerase chain reaction(PCR) was used to detect the expression of miR-130b and the host gene 2610318N02Rik (RIK); MPC5 were transfected with MiR-130b inhibitor or normal control(NC) inhibitor, then the cells were stimulated with 100 μg/mL PAN for 24 h, then western blot was used to detected the protein expression of Synaptopodin and Nephrin. Phalloidin dying was used to detect the changes of cytoskeleton. Flow cytometry was used to measure podocyte apoptosis. Targetscan 7.2 was adopted to predict the potential target gene of miR-130b, and Western blot was used to detect effect of miR-130b mimic on expression of the potential target gene PGC1a. Dual-luciferase reporter assay system was utilized to detect effect of miR-130b mimic on fluorescence activity. Results PAN significantly inhibited the expression of miR-130b and RIK. The western blot showed that inhibition of miR-130b increased the protein expression of Synaptopodin and Nephrin compared to NC inhibitor group. Phalloidin dying showed that inhibition of miR-130b partially alleviated cytoskeletal remodeling of podocytes induced by PAN. Flow-cytometric analysis showed that PAN-induced apoptosis was decreased after miR-130b silencing; Targetcan 7.2 analysis showed miR-130b mimic could significantly down-regulate the protein expression of PGC1α,the dual luciferase reporter assay system results showed that miR-130b and mild PGC1α 3'-UTR co-transfection decreased PGC1α 3'- UTR luciferase activity compared to the control mimic group and the mutant PGC1α 3'-UTR co-transfection group, but there was no significant difference between the control mimic group and the mutant ·PGC1α 3'- UTR group.Conclusions MiR-130b involves in podocyte injury induced by PAN by target inhibition of PGC1α expression.