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Calcitriol Ameliorates AngiotensinII-Induced Renal Injury Partly via Upregulating A20

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Abstract

Inflammation and reactive oxygen species (ROS) play crucial roles in the progression of chronic kidney diseases. Vitamin D has been shown anti-inflammatory effects, but the underlying mechanism is not fully understood. Here, we investigated whether calcitriol exerts protective effects via upregulating A20 in angiotensinII (AngII)-induced renal injury. Male C57BL/6 mice were infused with vehicle or AngII for 10 days. Calcitriol reduced infiltration of T lymphocytes and macrophages. This reduction of inflammatory cells was accompanied by elevated A20 and decreased pro-inflammatory cytokines (PICs) and reactive oxygen species (ROS). Calcitriol could inhibit NF-κB activation and necroptotic pathway. Induction of A20 was located primarily to the tubular epithelial cells. In rat proximal tubular epithelial cells (NRK-52E), calcitriol stably upregulated A20 and reduced the PICs and ROS. Inhibitory effect of A20 on PICs and ROS depended on suppressing NF-κB pathway and necroptotic pathway, respectively. A20 knockdown diminished the effect of calcitriol on suppressing NF-κB and necroptotic pathways. However, A20 deficiency could not abrogate the inhibitory effect of calcitriol on NF-κB and necroptotic pathways. Our results established that A20 is involved in the renoprotective effect by calcitriol via negatively modulating the NF-κB pathway and necroptotic pathway in AngII-induced renal injury.

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References

  1. Stenvinkel, P. 2006. Inflammation in end-stage renal disease: The hidden enemy. Nephrology (Carlton) 11 (1): 36–41. doi:10.1111/j.1440-1797.2006.00541.x.

    Article  Google Scholar 

  2. Holterman, C.E., N.C. Read, and C.R. Kennedy. 2015. Nox and renal disease. Clinical Science (London, England) 128 (8): 465–481. doi:10.1042/CS20140361.

    Article  CAS  Google Scholar 

  3. Ruiz-Ortega, M., M. Ruperez, O. Lorenzo, V. Esteban, J. Blanco, S. Mezzano, and J. Egido. 2002. Angiotensin II regulates the synthesis of proinflammatory cytokines and chemokines in the kidney. Kidney International. Supplement 82: S12–S22. doi:10.1046/j.1523-1755.62.s82.4.x.

    Article  CAS  Google Scholar 

  4. Kim, S.M., Y.G. Kim, K.H. Jeong, S.H. Lee, T.W. Lee, C.G. Ihm, and J.Y. Moon. 2012. Angiotensin II-induced mitochondrial Nox4 is a major endogenous source of oxidative stress in kidney tubular cells. PloS One 7 (7): e39739. doi:10.1371/journal.pone.0039739.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Jaattela, M., H. Mouritzen, F. Elling, and L. Bastholm. 1996. A20 zinc finger protein inhibits TNF and IL-1 signaling. Journal of Immunology 156 (3): 1166–1173.

    CAS  Google Scholar 

  6. Onizawa, M., S. Oshima, U. Schulze-Topphoff, J.A. Oses-Prieto, T. Lu, R. Tavares, T. Prodhomme, et al. 2015. The ubiquitin-modifying enzyme A20 restricts ubiquitination of the kinase RIPK3 and protects cells from necroptosis. Nature Immunology 16 (6): 618–627. doi:10.1038/ni.3172.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Verstrepen, L., K. Verhelst, G. van Loo, I. Carpentier, S.C. Ley, and R. Beyaert. 2010. Expression, biological activities and mechanisms of action of A20 (TNFAIP3). Biochemical Pharmacology 80 (12): 2009–2020. doi:10.1016/j.bcp.2010.06.044.

    Article  CAS  PubMed  Google Scholar 

  8. Wertz, I.E., K.M. O'Rourke, H. Zhou, M. Eby, L. Aravind, S. Seshagiri, P. Wu, et al. 2004. De-ubiquitination and ubiquitin ligase domains of A20 downregulate NF-kappaB signalling. Nature 430 (7000): 694–699. doi:10.1038/nature02794.

    Article  CAS  PubMed  Google Scholar 

  9. Wertz, I.E., K. Newton, D. Seshasayee, S. Kusam, C. Lam, J. Zhang, N. Popovych, et al. 2015. Phosphorylation and linear ubiquitin direct A20 inhibition of inflammation. Nature 528 (7582): 370–375. doi:10.1038/nature16165.

    Article  CAS  PubMed  Google Scholar 

  10. Lutz, J., A. Luongle, M. Strobl, M. Deng, H. Huang, M. Anton, M. Zakkar, et al. 2008. The A20 gene protects kidneys from ischaemia/reperfusion injury by suppressing pro-inflammatory activation. J Mol Med (Berl) 86 (12): 1329–1339. doi:10.1007/s00109-008-0405-4.

    Article  CAS  Google Scholar 

  11. Lee, E.G., D.L. Boone, S. Chai, S.L. Libby, M. Chien, J.P. Lodolce, and A. Ma. 2000. Failure to regulate TNF-induced NF-kappaB and cell death responses in A20-deficient mice. Science 289 (5488): 2350–2354.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Kunter, U., S. Daniel, M.B. Arvelo, J. Choi, T. Shukri, V.I. Patel, C.R. Longo, et al. 2005. Combined expression of A1 and A20 achieves optimal protection of renal proximal tubular epithelial cells. Kidney International 68 (4): 1520–1532. doi:10.1111/j.1523-1755.2005.00564.x.

    Article  CAS  PubMed  Google Scholar 

  13. Yang, Z., Z. Zhong, M. Li, Y. Xiong, Y. Wang, G. Peng, and Q. Ye. 2016. Hypothermic machine perfusion increases A20 expression which protects renal cells against ischemia/reperfusion injury by suppressing inflammation, apoptosis and necroptosis. International Journal of Molecular Medicine 38 (1): 161–171. doi:10.3892/ijmm.2016.2586.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Ketteler, M., G.J. Elder, P. Evenepoel, J.H. Ix, S.A. Jamal, M.H. Lafage-Proust, R. Shroff, et al. 2015. Revisiting KDIGO clinical practice guideline on chronic kidney disease-mineral and bone disorder: A commentary from a kidney disease: Improving global outcomes controversies conference. Kidney International 87 (3): 502–528. doi:10.1038/ki.2014.425.

    Article  PubMed  Google Scholar 

  15. Zhang, Y., J. Kong, D.K. Deb, A. Chang, and Y.C. Li. 2010. Vitamin D receptor attenuates renal fibrosis by suppressing the renin-angiotensin system. Journal of the American Society of Nephrology 21 (6): 966–973. doi:10.1681/ASN.2009080872.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Tan, X., X. Wen, and Y. Liu. 2008. Paricalcitol inhibits renal inflammation by promoting vitamin D receptor-mediated sequestration of NF-kappaB signaling. Journal of the American Society of Nephrology 19 (9): 1741–1752. doi:10.1681/ASN.2007060666.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Garcia, I.M., L. Altamirano, L. Mazzei, M. Fornes, M.N. Molina, L. Ferder, and W. Manucha. 2012. Role of mitochondria in paricalcitol-mediated cytoprotection during obstructive nephropathy. American Journal of Physiology. Renal Physiology 302 (12): F1595–F1605. doi:10.1152/ajprenal.00617.2011.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Riachy, R., B. Vandewalle, J. Kerr Conte, E. Moerman, P. Sacchetti, B. Lukowiak, V. Gmyr, T. Bouckenooghe, M. Dubois, and F. Pattou. 2002. 1,25-dihydroxyvitamin D3 protects RINm5F and human islet cells against cytokine-induced apoptosis: Implication of the antiapoptotic protein A20. Endocrinology 143 (12): 4809–4819. doi:10.1210/en.2002-220449.

    Article  CAS  PubMed  Google Scholar 

  19. Gregori, S., M. Casorati, S. Amuchastegui, S. Smiroldo, A.M. Davalli, and L. Adorini. 2001. Regulatory T cells induced by 1 alpha,25-dihydroxyvitamin D3 and mycophenolate mofetil treatment mediate transplantation tolerance. Journal of Immunology 167 (4): 1945–1953.

    Article  CAS  Google Scholar 

  20. Rice, E.K., G.H. Tesch, Z. Cao, M.E. Cooper, C.N. Metz, R. Bucala, R.C. Atkins, and D.J. Nikolic-Paterson. 2003. Induction of MIF synthesis and secretion by tubular epithelial cells: A novel action of angiotensin II. Kidney International 63 (4): 1265–1275. doi:10.1046/j.1523-1755.2003.00875.x.

    Article  CAS  PubMed  Google Scholar 

  21. Romero, D.G., M. Plonczynski, G.R. Vergara, E.P. Gomez-Sanchez, and C.E. Gomez-Sanchez. 2004. Angiotensin II early regulated genes in H295R human adrenocortical cells. Physiological Genomics 19 (1): 106–116. doi:10.1152/physiolgenomics.00097.2004.

    Article  CAS  PubMed  Google Scholar 

  22. Geilen, C.C., M. Bektas, T. Wieder, V. Kodelja, S. Goerdt, and C.E. Orfanos. 1997. 1alpha,25-dihydroxyvitamin D3 induces sphingomyelin hydrolysis in HaCaT cells via tumor necrosis factor alpha. The Journal of Biological Chemistry 272 (14): 8997–9001.

    Article  CAS  PubMed  Google Scholar 

  23. Zhu, Y., H. Cui, H. Gan, Y. Xia, L. Wang, Y. Wang, and Y. Sun. 2015. Necroptosis mediated by receptor interaction protein kinase 1 and 3 aggravates chronic kidney injury of subtotal nephrectomised rats. Biochemical and Biophysical Research Communications 461 (4): 575–581. doi:10.1016/j.bbrc.2015.03.164.

    Article  CAS  PubMed  Google Scholar 

  24. Wang, Y., D.K. Deb, Z. Zhang, T. Sun, W. Liu, D. Yoon, J. Kong, Y. Chen, A. Chang, and Y.C. Li. 2012. Vitamin D receptor signaling in podocytes protects against diabetic nephropathy. Journal of the American Society of Nephrology 23 (12): 1977–1986. doi:10.1681/ASN.2012040383.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Zhang, Z., W. Yuan, L. Sun, F.L. Szeto, K.E. Wong, X. Li, J. Kong, and Y.C. Li. 2007. 1,25-Dihydroxyvitamin D3 targeting of NF-kappaB suppresses high glucose-induced MCP-1 expression in mesangial cells. Kidney International 72 (2): 193–201. doi:10.1038/sj.ki.5002296.

    Article  CAS  PubMed  Google Scholar 

  26. Chen, Y., J. Zhang, X. Ge, J. Du, D.K. Deb, and Y.C. Li. 2013. Vitamin D receptor inhibits nuclear factor kappaB activation by interacting with IkappaB kinase beta protein. The Journal of Biological Chemistry 288 (27): 19450–19458. doi:10.1074/jbc.M113.467670.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. da Silva, C.G., E.R. Maccariello, S.W. Wilson, P. Putheti, S. Daniel, S.M. Damrauer, C.R. Peterson, J.J. Siracuse, E. Kaczmarek, and C. Ferran. 2012. Hepatocyte growth factor preferentially activates the anti-inflammatory arm of NF-kappaB signaling to induce A20 and protect renal proximal tubular epithelial cells from inflammation. Journal of Cellular Physiology 227 (4): 1382–1390. doi:10.1002/jcp.22851.

    Article  PubMed  PubMed Central  Google Scholar 

  28. He, K.L., and A.T. Ting. 2002. A20 inhibits tumor necrosis factor (TNF) alpha-induced apoptosis by disrupting recruitment of TRADD and RIP to the TNF receptor 1 complex in Jurkat T cells. Molecular and Cellular Biology 22 (17): 6034–6045.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Li, M., X. Shi, T. Qian, J. Li, Z. Tian, B. Ni, and F. Hao. 2015. A20 overexpression alleviates pristine-induced lupus nephritis by inhibiting the NF-kappaB and NLRP3 inflammasome activation in macrophages of mice. International Journal of Clinical and Experimental Medicine 8 (10): 17430–17440.

    PubMed  PubMed Central  Google Scholar 

  30. Skaug, B., J. Chen, F. Du, J. He, A. Ma, and Z.J. Chen. 2011. Direct, noncatalytic mechanism of IKK inhibition by A20. Molecular Cell 44 (4): 559–571. doi:10.1016/j.molcel.2011.09.015.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Zilberman-Rudenko, J., L.M. Shawver, A.W. Wessel, Y. Luo, M. Pelletier, W.L. Tsai, Y. Lee, et al. 2016. Recruitment of A20 by the C-terminal domain of NEMO suppresses NF-kappaB activation and autoinflammatory disease. Proceedings of the National Academy of Sciences of the United States of America 113 (6): 1612–1617. doi:10.1073/pnas.1518163113.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Kanamaru, Y., S. Sekine, H. Ichijo, and K. Takeda. 2012. The phosphorylation-dependent regulation of mitochondrial proteins in stress responses. Journal of Signal Transduction 2012: 931215. doi:10.1155/2012/931215.

    Article  PubMed  PubMed Central  Google Scholar 

  33. Zhang, D.W., J. Shao, J. Lin, N. Zhang, B.J. Lu, S.C. Lin, M.Q. Dong, and J. Han. 2009. RIP3, an energy metabolism regulator that switches TNF-induced cell death from apoptosis to necrosis. Science 325 (5938): 332–336. doi:10.1126/science.1172308.

    Article  CAS  PubMed  Google Scholar 

  34. Arora, D., P.K. Sharma, M.H. Siddiqui, and Y. Shukla. 2017. Necroptosis: Modules and molecular switches with therapeutic implications. Biochimie. doi:10.1016/j.biochi.2017.02.015.

  35. Li, X.H., X.P. Huang, L. Pan, C.Y. Wang, J. Qin, F.W. Nong, Y.Z. Luo, et al. 2016. Vitamin D deficiency may predict a poorer outcome of IgA nephropathy. BMC Nephrology 17 (1): 164. doi:10.1186/s12882-016-0378-4.

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

We thank Lixue Chen, WeixueTang, Jingmei Xie, Yao Xiao, Xiaojuan Deng, and Guangchen Qin at the central laboratory of the First Affiliated Hospital of Chongqing Medical University for their assistance during the experiments.

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Authors and Affiliations

  1. Department of Nephrology, The First Affiliated Hospital of Chongqing Medical University, No. 1 Youyi Road, Yuzhong District, Chongqing, 400016, China

    Hongfei Zhao, Yunfeng Xia & Hua Gan

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  1. Hongfei Zhao
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Authors’ Contributions

Hongfei Zhao conceived the study and designed the experiments. Hongfei Zhao performed all the experiments, analyzed the data, and wrote the manuscript. Hua Gan and Yunfeng Xia reviewed and revised the paper.

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Correspondence to Hua Gan.

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Zhao, H., Xia, Y. & Gan, H. Calcitriol Ameliorates AngiotensinII-Induced Renal Injury Partly via Upregulating A20. Inflammation 40, 1884–1893 (2017). https://doi.org/10.1007/s10753-017-0629-y

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