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Down-regulation of miR-186 contributes to podocytes apoptosis in membranous nephropathy
Wen-gang Sha1, Lei Shen1, Ling Zhou1
1Department of Nephrology, The First Affiliated Hospital of Soochow University, Suzhou 215006, PR China.
Background And Aim:
Podocytes apoptosis is the key process in the development of membranous nephropathy and miR-186 is reported to be related with cell apoptosis. Here we investigated the expression of miR-186 in membranous nephropathy (MGN) patients and the mechanism underlying the podocytes apoptosis.
Methods:
Thirty patients with MGN and 30 healthy people were included in this study. The expression of miR-186 was detected in renal tissue and podocyte cells exposed to AngII by real-time PCR. Caspase-3 activity was used to evaluate podocytes apoptosis. TLR4 and P2×7 protein expression was quantified by western blotting. miR-186 inhibitor and miR-186 mimic were transfected into cells to investigate the mechanism underlying miR-186 in podocytes apoptosis.
Results:
In MGN patients, the level of miR-186 was significantly down-regulated as well as the protein expression of TLR4 and P2×7 was up-regulated in renal tissue. In vitro experiments, TLR4 siRNA increased the expression of miR-186 and miR-186 inhibitor elevated the mRNA and protein expression of P2×7 in podocytes exposed to AngII. In addition, the level of cleaved-caspase-3 was up-regulated by miR-186 inhibitor. The TUNEL-positive cells and caspase-3 activity of podocytes induced by AngII were down-regulated by miR-186 mimic.
Conclusions:
We revealed that TLR4 is involved in the regulation of miR-186 expression, and the anti-apoptotic effect of miR-186 on podocytes is correlated with P2×7 regulation.
Insights
MicroRNA-186 (miR-186) is down-regulated in membranous nephropathy, contributing to podocyte apoptosis via P2×7 regulation. Restoring miR-186 levels may offer a therapeutic strategy for this kidney disease.
Area of Science:
- Nephrology
- Molecular Biology
- Cell Biology
Background:
- Podocyte apoptosis is central to membranous nephropathy (MGN) pathogenesis.
- MicroRNA-186 (miR-186) has been implicated in cellular apoptosis.
Purpose of the Study:
- To investigate miR-186 expression in MGN patients.
- To elucidate the mechanism of miR-186 in podocyte apoptosis.
Main Methods:
- Real-time PCR for miR-186 expression in renal tissue and AngII-exposed podocytes.
- Western blotting for TLR4 and P2×7 protein levels.
- Transfection with miR-186 inhibitor/mimic and TLR4 siRNA in podocytes.
Main Results:
- miR-186 was downregulated, while TLR4 and P2×7 were upregulated in MGN patients.
- TLR4 modulation affected miR-186 expression; miR-186 inhibition increased P2×7 and cleaved-caspase-3.
- miR-186 mimic reduced AngII-induced podocyte apoptosis.
Conclusions:
- TLR4 regulates miR-186 expression in podocytes.
- miR-186 exerts anti-apoptotic effects on podocytes by regulating P2×7.
