Abstract
Angiotensin II (Ang II) has been reported to cause podocyte apoptosis in rats both in vivo and in vitro studies. However, the underlying mechanisms are poorly understood. In the present study, we investigated the role of the nonreceptor tyrosine kinase c-Abl in Ang II-induced podocyte apoptosis. Male Sprague–Dawley rats in groups of 12 were administered either Ang II (400 kg/kg/min) or Ang II + STI-571 (50 mg/kg/day) by osmotic minipumps. In addition, 12 rats-receiving normal saline served as the control. Glomeruli c-Abl expression was carried out by real time PCR, Western blotting and immunolabeled, and occurrence of apoptosis was carried out by TUNEL staining and transmission electron microscopic analysis. In vitro studies, conditionally immortalized mouse podocytes were treated with Ang II (10−9–10−6 M) in the presence or absence of either c-Abl inhibitor, Src-I1, specific c-Abl siRNA, or c-Abl plasmid alone. Quantification of podocyte c-Abl expression and c-Abl phosphorylation at Y245 and Y412 was carried out by real time PCR, Western blotting and immunofluorescence imaging. The nuclear c-Abl and p53 were quantified by co-immunoprecipitation and Western blotting studies. Podocyte apoptosis was analysed by flow cytometry and Hoechst-33342 staining. c-Abl expression was demonstrated in rat kidney podocytes in vivo and cultured mouse podocytes in vitro. Ang II-receiving rats displayed enhanced podocyte c-Abl expression. And Ang II significantly stimulated c-Abl expression in cultured podocytes. Furthermore Ang II upregulated podocyte c-Abl phosphorylation at Y245 and Y412. Ang II also induced an increase of nuclear p53 protein and nuclear c-Abl-p53 complexes in podocytes and podocyte apoptosis. Down-regulation of c-Abl expression by c-Abl inhibitor (Src-I1) as well as specific siRNA inhibited Ang II-induced podocyte apoptosis; conversely, podoctyes transfected with c-Abl plasmid displayed enhanced apoptosis. These findings indicate that c-Abl may mediates Ang II-induced podocyte apoptosis, and inhibition of c-Abl expression can protect podocytes from Ang II-induced injury.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aoyama K, Fukumoto Y, Ishibashi K, Kubota S, Morinaga T, Horiike Y, Yuki R, Takahashi A, Nakayama Y, Yamaguchi N (2011) Nuclear c-Abl-mediated tyrosine phosphorylation induces chromatin structural changes through histone modifications that include H4K16 hypoacetylation. Exp Cell Res 317:2874–2903
Bain J, Plater L, Elliott M, Shpiro N, Hastie CJ, McLauchlan H, Klevernic I, Arthur JS, Alessi DR, Cohen P (2007) The selectivity of protein kinase inhibitors: a further update. Biochem J 408:297–315
Bradley WD, Koleske AJ (2009) Regulation of cell migration and morphogenesis by Abl-family kinases: emerging mechanisms and physiological contexts. J Cell Sci 122:3441–3454
Colicelli J (2010) ABL tyrosine kinases: evolution of function, regulation, and specificity. Sci Signal 3:e6
Ding G, Reddy K, Kapasi AA, Franki N, Gibbons N, Kasinath BS, Singhal PC (2002) Angiotensin II induces apoptosis in rat glomerular epithelial cells. Am J Physiol Renal Physiol 283:F173–F180
Eid AA, Ford BM, Block K, Kasinath BS, Gorin Y, Ghosh-Choudhury G, Barnes JL, Abboud HE (2010) AMP-activated protein kinase (AMPK) negatively regulates Nox4-dependent activation of p53 and epithelial cell apoptosis in diabetes. J Biol Chem 285:37503–37512
Furlan A, Stagni V, Hussain A, Richelme S, Conti F, Prodosmo A, Destro A, Roncalli M, Barila D, Maina F (2011) Abl interconnects oncogenic Met and p53 core pathways in cancer cells. Cell Death Differ 18:1608–1616
Jia DY, Du ZH, Liu SM, Liu HJ, Wang FW, Ling EA, Liu K, Hao AJ (2008a) c-Abl is involved in high glucose-induced apoptosis in embryonic E12.5 cortical neural progenitor cells from the mouse brain. J Neurochem 106:1720–1730
Jia J, Ding G, Zhu J, Chen C, Liang W, Franki N, Singhal PC (2008b) Angiotensin II infusion induces nephrin expression changes and podocyte apoptosis. Am J Nephrol 28:500–507
Jung DS, Lee SH, Kwak SJ, Li JJ, Kim DH, Nam BY, Kang HY, Chang TI, Park JT, Han SH et al (2012) Apoptosis occurs differentially according to glomerular size in diabetic kidney disease. Nephrol Dial Transplant 27:259–266
Klein A, Maldonado C, Vargas LM, Gonzalez M, Robledo F, Perez DAK, Munoz FJ, Hetz C, Alvarez AR, Zanlungo S (2011) Oxidative stress activates the c-Abl/p73 proapoptotic pathway in Niemann-Pick type C neurons. Neurobiol Dis 41:209–218
Liu Q, Wang G, Zhou G, Tan Y, Wang X, Wei W, Liu L, Xue W, Feng W, Cai L (2009) Angiotensin II-induced p53-dependent cardiac apoptotic cell death: its prevention by metallothionein. Toxicol Lett 191:314–320
Megyeri K, Orosz L, Kemeny L (2007) Vesicular stomatitis virus infection triggers apoptosis associated with decreased DeltaNp63alpha and increased Bax levels in the immortalized HaCaT keratinocyte cell line. Biomed Pharmacother 61:254–260
Nicholas SB, Basgen JM, Sinha S (2011) Using stereologic techniques for podocyte counting in the mouse: shifting the paradigm. Am J Nephrol 33(Suppl 1):1–7
Qiu Z, Cang Y, Goff SP (2010) Abl family tyrosine kinases are essential for basement membrane integrity and cortical lamination in the cerebellum. J Neurosci 30:14430–14439
Rafalska I, Zhang Z, Benderska N, Wolff H, Hartmann AM, Brack-Werner R, Stamm S (2004) The intranuclear localization and function of YT521-B is regulated by tyrosine phosphorylation. Hum Mol Genet 13:1535–1549
Ren Z, Liang W, Chen C, Yang H, Singhal PC, Ding G (2012) Angiotensin II induces nephrin dephosphorylation and podocyte injury: role of caveolin-1. Cell Signal 24:443–450
Saha MN, Jiang H, Chang H (2010) Molecular mechanisms of nutlin-induced apoptosis in multiple myeloma: evidence for p53-transcription-dependent and -independent pathways. Cancer Biol Ther 10:567–578
Sanchez-Nino MD, Sanz AB, Sanchez-Lopez E, Ruiz-Ortega M, Benito-Martin A, Saleem MA, Mathieson PW, Mezzano S, Egido J, Ortiz A (2012) HSP27/HSPB1 as an adaptive podocyte antiapoptotic protein activated by high glucose and angiotensin II. Lab Invest 92:32–45
Sanwal V, Pandya M, Bhaskaran M, Franki N, Reddy K, Ding G, Kapasi A, Valderrama E, Singhal PC (2001) Puromycin aminonucleoside induces glomerular epithelial cell apoptosis. Exp Mol Pathol 70:54–64
Schlatterer SD, Tremblay MA, Acker CM, Davies P (2011) Neuronal c-Abl overexpression leads to neuronal loss and neuroinflammation in the mouse forebrain. J Alzheimers Dis 25:119–133
Skirnisdottir I, Seidal T (2012) The apoptosis regulators p53, bax and PUMA: relationship and impact on outcome in early stage (FIGO I-II) ovarian carcinoma after post-surgical taxane-based treatment. Oncol Rep 27:741–747
Tuncdemir M, Ozturk M (2011) The effects of angiotensin-II receptor blockers on podocyte damage and glomerular apoptosis in a rat model of experimental streptozotocin-induced diabetic nephropathy. Acta Histochem 113:826–832
Wada T, Pippin JW, Marshall CB, Griffin SV, Shankland SJ (2005) Dexamethasone prevents podocyte apoptosis induced by puromycin aminonucleoside: role of p53 and Bcl-2-related family proteins. J Am Soc Nephrol 16:2615–2625
Wada T, Pippin JW, Nangaku M, Shankland SJ (2008) Dexamethasone’s prosurvival benefits in podocytes require extracellular signal-regulated kinase phosphorylation. Nephron Exp Nephrol 109:e8–e19
Wang S, Wilkes MC, Leof EB, Hirschberg R (2010) Noncanonical TGF-beta pathways, mTORC1 and Abl, in renal interstitial fibrogenesis. Am J Physiol Renal Physiol 298:F142–F149
Wang X, Zeng L, Wang J, Chau JF, Lai KP, Jia D, Poonepalli A, Hande MP, Liu H, He G et al (2011) A positive role for c-Abl in Atm and Atr activation in DNA damage response. Cell Death Differ 18:5–15
Westlund BS, Cai B, Zhou J, Sparrow JR (2009) Involvement of c-Abl, p53 and the MAP kinase JNK in the cell death program initiated in A2E-laden ARPE-19 cells by exposure to blue light. Apoptosis 14:31–41
Xu J, Millard M, Ren X, Cox OT, Erdreich-Epstein A (2010) c-Abl mediates endothelial apoptosis induced by inhibition of integrins alphavbeta3 and alphavbeta5 and by disruption of actin. Blood 115:2709–2718
Yadav A, Vallabu S, Arora S, Tandon P, Slahan D, Teichberg S, Singhal PC (2010) ANG II promotes autophagy in podocytes. Am J Physiol Cell Physiol 299:C488–C496
Zhou LL, Cao W, Xie C, Tian J, Zhou Z, Zhou Q, Zhu P, Li A, Liu Y, Miyata T et al (2012a) The receptor of advanced glycation end products plays a central role in advanced oxidation protein products-induced podocyte apoptosis. Kidney Int 82:759–770
Zhou Y, Zhang E, Berggreen C, Jing X, Osmark P, Lang S, Cilio CM, Goransson O, Groop L, Renstrom E et al (2012b) Survival of pancreatic beta cells is partly controlled by a TCF7L2-p53-p53INP1-dependent pathway. Hum Mol Genet 21:196–207
Zhuo JL, Li XC (2011) New insights and perspectives on intrarenal renin-angiotensin system: focus on intracrine/intracellular angiotensin II. Peptides 32:1551–1565
Acknowledgments
These studies were supported by grants from the National Science Foundation of China (30871167, 81270762 to G.D., 81100478 to W.L. and 30900688 to C.C.) and from the National Institutes of Health (RO1DK084910 to P.C.S).
Conflict of interest
The authors declare that they have no competing interests. The authors alone are responsible for the content and writing of the paper.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Chen, X., Ren, Z., Liang, W. et al. c-Abl mediates angiotensin II-induced apoptosis in podocytes. J Mol Hist 44, 597–608 (2013). https://doi.org/10.1007/s10735-013-9505-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10735-013-9505-8