Role and its mechanism of micro RNA-130b in puromycin aminonucleoside-induced podocyte injury
Objective To investigate the role and its mechanism of miR-130 b 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 express...
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| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | zho |
| Published: |
Editorial Department of Journal of Clinical Nephrology
2020-01-01
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| Series: | Linchuang shenzangbing zazhi |
| Subjects: | |
| Online Access: | http://www.lcszb.com/thesisDetails?columnId=57907776&Fpath=home&index=0 |
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| Summary: | Objective To investigate the role and its mechanism of miR-130 b 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-130 b and the host gene 2610318 N02 Rik(RIK); MPC5 were transfected with MiR-130 b 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-130 b, and Western blot was used to detect effect of miR-130 b mimic on expression of the potential target gene PGC1 a. Dual-luciferase reporter assay system was utilized to detect effect of miR-130 b mimic on fluorescence activity. Results PAN significantly inhibited the expression of miR-130 b and RIK. The western blot showed that inhibition of miR-130 b increased the protein expression of Synaptopodin and Nephrin compared to NC inhibitor group. Phalloidin dying showed that inhibition of miR-130 b partially alleviated cytoskeletal remodeling of podocytes induced by PAN. Flow-cytometric analysis showed that PAN-induced apoptosis was decreased after miR-130 b silencing; Targetcan 7.2 analysis showed miR-130 b mimic could significantly down-regulate the protein expression of PGC1α,the dual luciferase reporter assay system results showed that miR-130 b 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-130 b involves in podocyte injury induced by PAN by target inhibition of PGC1α expression. |
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| ISSN: | 1671-2390 |