Abstract
Aim:
NLRP3 inflammasome plays an important role in renal injury and may be a therapeutic target in the treatment of patients with progressive chronic kidney disease. In this study we investigated whether angiotensin II (Ang II)-induced NLRP3 inflammasome activation was linked to endoplasmic reticulum stress (ERS) in human renal proximal tubular cells in vitro.
Methods:
Human kidney proximal epithelial cells (HK-2) were pretreated with telmisartan or 4-PBA, and then treated with Ang II. The expression levels of mRNAs and proteins related to NLRP3 inflammasomes and ERS was examined by real-time PCR, Western blot and immunofluorescence.
Results:
Treatment with Ang II (10, 100, and 1000 nmol/L) increased the expression of the inflammasome markers NLRP3 and ASC, as well as caspase-1, IL-1β, and IL-18 in dose- and time-dependent manners with peak levels detected at 100 nmol/L and 12 h. Ang II-induced increases in the expression of NLRP3, ASC, caspase-1, IL-1β, and IL-18 were significantly reduced by pretreatment with telmisartan (1 μmol/L). Immunofluorescence studies showed that Ang II increased the expression of NLRP3 and ASC, which was inhibited by telmisartan. Furthermore, Ang II treatment increased the expression of ERS markers GRP78 and p-eIF2α in dose- and time-dependent manners, which was significantly reduced by telmisartan. Moreover, Ang II-induced increases in the expression of NLRP3, ASC, caspase-1, IL-1β, and IL-18 were significantly inhibited by pretreatment with the ERS inhibitor 4-PBA (5 mmol/L).
Conclusion:
Ang II treatment induces NLRP3 inflammasome activation in HK-2 cells in vitro and ER stress is involved in this process, which may represent a new mechanism for the renal rennin-angiotensin system to induce tubulointerstitial inflammation.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Chuang PY, Menon MC, He JC . Molecular targets for treatment of kidney fibrosis. J Mol Med 2013; 91: 549–59.
Silverstein DM . Inflammation in chronic kidney disease: role in the progression of renal and cardiovascular disease. Pediatr Nephrol 2009; 24: 1445–52.
Kobori H, Nangaku M, Navar LG, Nishiyama A . The intrarenal renin-angiotensin system: from physiology to the pathobiology of hypertension and kidney disease. Pharmacol Rev 2007; 59: 251–87.
Ruster C, Wolf G . Renin-angiotensin-aldosterone system and progression of renal disease. J Am Soc Nephrol 2006; 17: 2985–91.
Lin L, Phillips WE, Manning RD . Intrarenal angiotensin II is associated with inflammation, renal damage, and dysfunction in dahl salt-sensitive hypertension. J Am Soc Hypertens 2009; 3: 306–14.
Alique M, Civantos E, Sanchez Lopez E, Lavoz C, Rayego Mateos S, Rodrigues Díez R, et al. Integrin-linked kinase plays a key role in the regulation of angiotensin II-induced renal inflammation. Clin Sci 2014; 127: 19–31.
Xie P, Joladarashi D, Dudeja P, Sun L, Kanwar YS . Modulation of angiotensin II-induced inflammatory cytokines by the Epac1-Rap1A-NHE3 pathway: implications in renal tubular pathobiology. Am J Physiol Renal Physiol 2014; 306: F1260–74.
Menu P, Mayor A, Zhou R, Tardivel A, Ichijo H, Mori K, et al. ER stress activates the NLRP3 inflammasome via an UPR-independent pathway. Cell Death Dis 2012; 3: e261.
Bauernfeind F, Ablasser A, Bartok E, Kim S, Schmid-Burgk J, Cavlar T, et al. Inflammasomes: current understanding and open questions. Cell Mol Life Sci 2011; 68: 765–83.
Vilaysane A, Chun J, Seamone ME, Wang W, Chin R, Hirota S, et al. The NLRP3 inflammasome promotes renal inflammation and contributes to CKD. J Am Soc Nephrol 2010; 21: 1732–44.
Schroder K, Tschopp J . The Inflammasomes. Cell 2010; 140: 821–32.
Ding LH, Liu D, Xu M, Liu H, Wu M, Tang RN, et al. Enalapril inhibits tubulointerstitial inflammation and NLRP3 inflammasome expression in BSA-overload nephropathy of rats. Acta Pharmacol Sin 2014; 35: 1293–301.
Ruiz-Ortega M, Ruperez M, Esteban V, Rodriguez-Vita J, Sanchez-Lopez E, Carvajal G, et al. Angiotensin II: a key factor in the inflammatory and fibrotic response in kidney diseases. Nephrol Dial Transpl 2005; 21: 16–20.
Ruiz-Ortega M, Bustos C, Hernandez-Presa MA, Lorenzo O, Plaza JJ, Egido J . Angiotensin II participates in mononuclear cell recruitment in experimental immune complex nephritis through nuclear factor-kappa B activation and monocyte chemoattractant protein-1 synthesis. J Immunol 1998; 161: 430–9.
Rafiq K, Nishiyama A, Konishi Y, Morikawa T, Kitabayashi C, Kohno M, et al. Regression of glomerular and tubulointerstitial injuries by dietary salt reduction with combination therapy of angiotensin II receptor blocker and calcium channel blocker in dahl salt-sensitive rats. PLoS One 2014; 9: e107853.
Ruiz-Ortega M, Lorenzo O, Suzuki Y, Rupérez M, Egido J . Proinflammatory actions of angiotensins. Curr Opin Nephrol Hypertens 2001; 10: 321–9.
Liu G, Li Y, Huang XR, Wei L, Chen H, Shi Y, et al. Disruption of Smad7 promotes ANG II-mediated renal inflammation and fibrosis via Sp1-TGF-β/Smad3-NFκB-dependent mechanisms in mice. PLoS One 2013; 8: e53573.
Dikalov SI, Nazarewicz RR . Angiotensin II-induced production of mitochondrial reactive oxygen species: potential mechanisms and relevance for cardiovascular disease. Antioxid Redox Signal 2013; 19: 1085–94.
Lorenz G, Darisipudi MN, Anders HJ . Canonical and non-canonical effects of the NLRP3 inflammasome in kidney inflammation and fibrosis. Nephrol Dial Transpl 2014; 29: 41–8.
Anders HJ, Muruve DA . The inflammasomes in kidney disease. J Am Soc Nephrol 2011; 22: 1007–18.
Darisipudi MN, Thomasova D, Mulay SR, Brech D, Noessner E, Liapis H, et al. Uromodulin triggers IL-1-dependent innate immunity via the NLRP3 inflammasome. J Am Soc Nephrol 2012; 23: 1783–9.
Mulay SR, Kulkarni OP, Rupanagudi KV, Migliorini A, Darisipudi MN, Vilaysane A, et al. Calcium oxalate crystals induce renal inflammation by NLRP3-mediated IL-1β secretion. J Clin Invest 2013; 123: 236–46.
Kim S, Joe Y, Jeong SO, Zheng M, Back SH, Park SW, et al. Endoplasmic reticulum stress is sufficient for the induction of IL-1beta production via activation of the NF-kappaB and inflammasome pathways. Innate Immun 2014; 20: 799–815.
Oslowski CM, Hara T, O'Sullivan-Murphy B, Kanekura K, Lu S, Hara M, et al. Thioredoxin-interacting protein mediates ER stress-induced beta cell death through initiation of the inflammasome. Cell Metab 2012; 16: 265–73.
Koyama M, Furuhashi M, Ishimura S, Mita T, Fuseya T, Okazaki Y, et al. Reduction of endoplasmic reticulum stress by 4-phenylbutyric acid prevents the development of hypoxia-induced pulmonary arterial hypertension. Am J Physiol Heart Circ Physiol 2014; 306: H1314–23.
Liu B, Gao J, Li Q, Xu L . Albumin caused the increasing production of angiotensin II due to the dysregulation of ACE/ACE2 expression in HK2 cells. Clin Chim Acta 2009; 403: 23–30.
Fang L, Xie D, Wu X, Cao H, Su W, Yang J . Involvement of endoplasmic reticulum stress in albuminuria induced inflammasome activation in renal proximal tubular cells. PLoS One 2013; 8: e72344.
Shigeoka AA, Mueller JL, Kambo A, Mathison JC, King AJ, Hall WF, et al. An inflammasome-independent role for epithelial-expressed Nlrp3 in renal ischemia-reperfusion injury. J Immunol 2010; 185: 6277–85.
Acknowledgements
This study was supported by grants from the National Natural Scientific Foundation (No 81130010), the Major State Basic Research Development Program (“973”) (No 2012CB517706), and the Program for Jiangsu Clinical Research Center (No BL2014080).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wang, J., Wen, Y., Lv, Ll. et al. Involvement of endoplasmic reticulum stress in angiotensin II-induced NLRP3 inflammasome activation in human renal proximal tubular cells in vitro. Acta Pharmacol Sin 36, 821–830 (2015). https://doi.org/10.1038/aps.2015.21
Received:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/aps.2015.21
Keywords
This article is cited by
-
Role of ER Stress Mediated Unfolded Protein Responses and ER Stress Inhibitors in the Pathogenesis of Inflammatory Bowel Disease
Digestive Diseases and Sciences (2022)
-
HDAC2 targeting stabilizes the CoREST complex in renal tubular cells and protects against renal ischemia/reperfusion injury
Scientific Reports (2021)
-
Hirsutella sinensis inhibits NLRP3 inflammasome activation to block aristolochic acid-induced renal tubular epithelial cell transdifferentiation
Human Cell (2020)
-
Caspase-11 promotes renal fibrosis by stimulating IL-1β maturation via activating caspase-1
Acta Pharmacologica Sinica (2019)