BACKGROUND: Angiotensin (Ang)II is involved in induction of proteinuria, renal injury, and apoptosis and thus a major contributor to the development of chronic kidney disease. Podocytes are of major importance for the pathogenesis of several kidney diseases. Decrease of podoplanin (PDPN) in podocytes and podocyte loss has been associated with the development of proteinuria. Little is known about the regulation and biological function of PDPN in podocytes and its role in AngII-mediated kidney damage. Here, we determined the influence of AngII on the expression of PDPN, microRNA (miRNA)-29b and miRNA-497 in human podocytes. Further, we analyzed the impact of small interfering RNA-mediated downregulation of PDPN on AngII-induced apoptosis and viability. Moreover, we characterized the role of miRNA-29b and miRNA-497 in expression regulation of PDPN. METHODS: Cell viability and apoptosis were determined by functional assays. Expression analyses were done via Real-Time PCR and western blot analyses. Dual luciferase assay was performed to characterize miRNA-mediated expression control. RESULTS: AngII increased the expression of miRNA-29b and reduced PDPN. Small interfering RNA-mediated downregulation of PDPN increased proapoptotic caspase-3 activation and cytochrome C translocation, whereas cell viability and Akt phosphorylation were reduced in AngII-stimulated podocytes. In contrast to miRNA-497, transfection of cells with miRNA-29b mimics significantly decreased PDPN. Cotransfection of cells with miRNA-29b and a dual luciferase reporter vector decreased the luciferase activity compared with controls. CONCLUSION: These data demonstrate the posttranscriptional control of PDPN expression by miRNA-29b and support a role of PDPN as an antiapoptotic prosurvival factor in AngII-induced injury of human podocytes.
BACKGROUND: Angiotensin (Ang)II is involved in induction of proteinuria, renal injury, and apoptosis and thus a major contributor to the development of chronic kidney disease. Podocytes are of major importance for the pathogenesis of several kidney diseases. Decrease of podoplanin (PDPN) in podocytes and podocyte loss has been associated with the development of proteinuria. Little is known about the regulation and biological function of PDPN in podocytes and its role in AngII-mediated kidney damage. Here, we determined the influence of AngII on the expression of PDPN, microRNA (miRNA)-29b and miRNA-497 in human podocytes. Further, we analyzed the impact of small interfering RNA-mediated downregulation of PDPN on AngII-induced apoptosis and viability. Moreover, we characterized the role of miRNA-29b and miRNA-497 in expression regulation of PDPN. METHODS: Cell viability and apoptosis were determined by functional assays. Expression analyses were done via Real-Time PCR and western blot analyses. Dual luciferase assay was performed to characterize miRNA-mediated expression control. RESULTS: AngII increased the expression of miRNA-29b and reduced PDPN. Small interfering RNA-mediated downregulation of PDPN increased proapoptotic caspase-3 activation and cytochrome C translocation, whereas cell viability and Akt phosphorylation were reduced in AngII-stimulated podocytes. In contrast to miRNA-497, transfection of cells with miRNA-29b mimics significantly decreased PDPN. Cotransfection of cells with miRNA-29b and a dual luciferase reporter vector decreased the luciferase activity compared with controls. CONCLUSION: These data demonstrate the posttranscriptional control of PDPN expression by miRNA-29b and support a role of PDPN as an antiapoptotic prosurvival factor in AngII-induced injury of human podocytes.
Authors: Angela Schulz; Nicola Victoria Müller; Monika Stoll; Reinhold Kreutz; Nina Anne van de Lest; Andreas Eisenreich; Martina Schmidbauer; Andrei Barysenka; Bettina Purfürst; Anje Sporbert; Theodor Lorenzen; Alexander M Meyer; Laura Herlan; Anika Witten; Frank Rühle; Weibin Zhou; Emile de Heer; Marion Scharpfenecker; Daniela Panáková Journal: Elife Date: 2019-03-22 Impact factor: 8.140