Abstract
Family focal segmental glomerulosclerosis (FSGS) is characterized by sclerosis and hyalinosis of particular loops of glomeruli and is one of the causes of the nephrotic syndrome. Certain mutations in the structure of TRPC6 channels are the genetic impetus for FSGS development resulting in podocytes functional abnormalities and various nephropathies. We have recently demonstrated that non-steroid antiinflammatory drugs (NSAID) ibuprofen and diclofenac decrease the activity of endogenous TRPC-like calcium channels in the podocytes of the freshly isolated rat glomeruli. It has also been shown that TRPC6 channels are expressed in the podocytes. In the current study we have functionally reconstituted TRPC6 channels in mammalian cells to investigate the effects of diclofenac on the activity of wild type TRPC6 channel and TRPC6P112Q channel containing a mutation in the N-terminus that was described in FSGS patients. Intracellular calcium level measurements in transfected cells revealed a more intensive carbachol-induced increase of calcium concentration in HEK-293 cells expressing TRPC6P112Q versus the cells expressing wild-type TRPC6. We also performed patch-clamp experiments to study TRPC6 channels reconstituted in Chinese hamster ovary (CHO) cell line and found that application of diclofenac (500 μM) acutely reduced single channel activity. Preincubation with diclofenac (100 μM) also decreased the whole-cell current in CHO cells overexpressing TRPC6P112Q. Therefore, our previously published data on the effects of NSAID on TRPC-like channels in the isolated rat glomeruli, along with this current investigation on the cultured overexpressed mammalian cells, allows hypothesizing that TRPC6 channels may be a target for NSAID that can be important in the treatment of FSGS.
Similar content being viewed by others
References
Shankland S.J. 2006. The podocyte’s response to injury: Role in proteinuria and glomerulosclerosis. Kidney Int. 69, 2131–2147.
Heeringa S.F., Moller C.C., Du J., Yue L., Hinkes B., Chernin G., Vlangos C.N., Hoyer P.F., Reiser J., Hildebrandt F. 2009. A novel TRPC6 mutation that causes childhood FSGS. PLoS ONE. 4, e7771.
Reiser J., Polu K.R., Moller C.C., Kenlan P., Altintas M.M., Wei C., Faul C., Herbert S., Villegas I., vila-Casado C., McGee, M., Sugimoto H., Brown D., Kalluri R., Mundel P., Smith P.L., Clapham D.E., Pollak M.R. 2005. TRPC6 is a glomerular slit diaphragm-associated channel required for normal renal function. Nat. Genet. 37, 739–744.
Winn M.P., Conlon P.J., Lynn K.L., Farrington M.K., Creazzo T., Hawkins A.F., Daskalakis N., Kwan S.Y., Ebersviller S., Burchette J.L., Pericak-Vance M.A., Howell D.N., Vance J.M., Rosenberg P.B. 2005. A mutation in the TRPC6 cation channel causes familial focal segmental glomerulosclerosis. Science. 308, 1801–1804.
De Miguel C., Lund H., Mattson D.L. 2011. High dietary protein exacerbates hypertension and renal damage in Dahl SS rats by increasing infiltrating immune cells in the kidney. Hypertension. 57, 269–274.
Haque M.Z., Ares G.R., Caceres P.S., Ortiz P.A. 2011. High salt differentially regulates surface NKCC2 expression in thick ascending limbs of Dahl salt-sensitive and salt-resistant rats. Am. J. Physiol. Renal Physiol. 300, F1096–F1104.
Hirano T., Ebara T., Furukawa S., Nagano S., Takahashi T. 1994. Mechanism of hypertriglyceridemia in dahl salt-sensitive rats, an animal model of spontaneous nephrotic syndrome. Metabolism. 43, 248–256.
Liu Y., Taylor N.E., Lu L., Usa K., Cowley A.W., Ferreri N.R., Yeo N.C., Liang M. 2010. Renal medullary microRNAs in Dahl salt-sensitive rats. Hypertension. 55, 974–982.
Kooijmans-Coutinho M.F., Tegzess A.M., Bruijn J.A., Florijn K.W., van Es L.A., van der Woude F.J. 1993. Indomethacin treatment of recurrent nephrotic syndrome and focal segmental glomerulosclerosis after renal transplantation. Nephrol. Dialysis Transplantation. 8, 469–473.
McCarthy E.T., Sharma M. 2002. Indomethacin protects permeability barrier from focal segmental glomerulosclerosis serum. Kidney Int. 61, 534–541.
Dorofeeva N.A., Barygin O.I., Staruschenko A., Bolshakov K.V., Magazanik L.G. 2008. Mechanisms of non-steroid anti-inflammatory drugs action on ASICs expressed in hippocampal interneurons. J. Neurochem. 106, 429–441.
Lee H.M., Kim H.I., Shin Y.K., Lee C.S., Park M., Song J.H. 2003. Diclofenac inhibition of sodium currents in rat dorsal root ganglion neurons. Brain Res. 992, 120–127.
Peretz A., Degani N., Nachman R., Uziyel Y., Gibor G., Shabat D., Attali B. 2005. Meclofenamic acid and diclofenac, novel templates of KCNQ2/Q3 potassium channel openers, depress cortical neuron activity and exhibit anticonvulsant properties. Mol. Pharmacol. 67, 1053–1066.
Pavlov T.S., Ilatovskaya D.V., Levchenko V., Mattson D.L., Roman R.J., Staruschenko A. 2011. Effects of cytochrome P450 metabolites of arachidonic acid on the epithelial sodium channel (ENaC). Am. J. Physiol. Renal Physiol. 301, F672–F681.
Li J., Xiang Y.Y., Ye L., Tsui L.C., MacDonald J.F., Hu J., Lu W.Y. 2008. Nonsteroidal anti-inflammatory drugs upregulate function of wild-type and mutant CFTR. Europ. Respir. J. 32, 334–343.
Gu R.M., Wang W.H. 2002. Arachidonic acid inhibits K channels in basolateral membrane of the thick ascending limb. Am. J. Physiol. Renal Physiol. 283, F407–F414.
Ilatovskaya D.V., Levchenko V., Ryan R.P., Cowley J., Staruschenko A. 2011. NSAIDs acutely inhibit TRPC channels in freshly isolated rat glomeruli. Biochem. Biophys. Res. Commun. 408, 242–247.
Goel M., Sinkins W.G., Zuo C.D., Estacion M., Schilling W.P. 2006. Identification and localization of TRPC channels in the rat kidney. Am. J. Physiol. Renal Physiol. 290, F1241–F1252.
Kim E.Y., varez-Baron C.P., Dryer S.E. 2009. Canonical transient receptor potential channel (TRPC) 3 and TRPC6 associate with large-conductance Ca2+-activated K+ (BKCa) channels: Role in BKCa trafficking to the surface of cultured Podocytes. Mol. Pharmacol. 75, 466–477.
Cowley A.W., Jr., Roman R.J., Kaldunski M.L., Dumas P., Dickhout J.G., Greene A.S., Jacob H.J. 2001. Brown Norway chromosome 13 confers protection from high salt to consomic Dahl S rat. Hypertension. 37, 456–461.
Mattson D.L., Dwinell M.R., Greene A.S., Kwitek A.E., Roman R.J., Jacob H.J., Cowley A.W., Jr. 2008. Chromosome substitution reveals the genetic basis of Dahl salt-sensitive hypertension and renal disease. Am. J. Physiol. Renal Physiol. 295, F837–F842.
Polichnowski A.J., Cowley A.W., Jr. 2009. Pressureinduced renal injury in angiotensin II versus norepinephrine-induced hypertensive rats. Hypertension. 54, 1269–1277.
Bal M., Zhang J., Hernandez C.C., Zaika O., Shapiro M.S. 2010. Ca2+/calmodulin disrupts AKAP79/150 interactions with KCNQ (M-type) K+ channels. J. Neurosci. 30, 2311–2323.
Graham S., Ding M., Sours-Brothers S., Yorio T., Ma J.X., Ma R. 2007. Downregulation of TRPC6 protein expression by high glucose, a possible mechanism for the impaired Ca2+ signaling in glomerular mesangial cells in diabetes. Am. J. Physiol. Renal Physiol. 293, F1381–F1390.
Graham S., Ding M., Ding Y., Sours-Brothers S., Luchowski R., Gryczynski Z., Yorio T., Ma H., Ma R. 2010. Canonical transient receptor potential 6 (TRPC6), a redox-regulated cation channel. J. Biol. Chem. 285, 23466–23476.
Grynkiewicz G., Poenie M., Tsien R.Y. 1985. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J. Biol. Chem. 260, 3440–3450.
Cayouette S., Lussier M.P., Mathieu E.L., Bousquet S.M., Boulay G. 2004. Exocytotic insertion of TRPC6 channel into the plasma membrane upon Gq Protein-coupled receptor activation. J. Biol. Chem. 279, 7241–7246.
Estacion M., Sinkins W.G., Jones S.W., Applegate M.A.B., Schilling W.P. 2006. Human TRPC6 expressed in HEK 293 cells forms non-selective cation channels with limited Ca2+ permeability. J. Physiol. 572, 359–377.
Shi J., Takahashi S., Jin X.H., Li Y.Q., Ito Y., Mori Y., Inoue R. 2007. Myosin light chain kinase-independent inhibition by ML-9 of murine TRPC6 channels expressed in HEK293 cells. Br. J. Pharmacol. 152, 122–131.
Hofmann T., Obukhov A.G., Schaefer M., Harteneck C., Gudermann T., Schultz G. 1999. Direct activation of human TRPC6 and TRPC3 channels by diacylglycerol. Nature. 397, 259–263.
Spernath A., Aserin A., Ziserman L., Danino D., Garti N. 2007. Phosphatidylcholine embedded microemulsions: Physical properties and improved Caco-2 cell permeability. J. Control Release. 119, 279–290.
Eckel J., Lavin P.J., Finch E.A., Mukerji N., Burch J., Gbadegesin R., Wu G., Bowling B., Byrd A., Hall G., Sparks M., Zhang Z.S., Homstad A., Barisoni L., Birbaumer L., Rosenberg P., Winn M.P. 2011. TRPC6 enhances angiotensin II-induced albuminuria. J. Am. Soc. Nephrol. 22, 526–535.
Hoenderop J.G., Voets T., Hoefs S., Weidema F., Prenen J., Nilius B., Bindels R.J. 2003. Homoand heterotetrameric architecture of the epithelial Ca2+ channels TRPV5 and TRPV6. EMBO J. 22, 776–785.
Kottgen M., Buchholz B., Garcia-Gonzalez M.A., Kotsis F., Fu X., Doerken M., Boehlke C., Steffl D., Tauber R., Wegierski T., Nitschke R., Suzuki M., Kramer-Zucker A., Germino G.G., Watnick T., Prenen J., Nilius B., Kuehn E.W., Walz G. 2008. TRPP2 and TRPV4 form a polymodal sensory channel complex. J. Cell Biol. 182, 437–447.
Schaefer M. 2005. Homoand heteromeric assembly of TRP channel subunits. Pflügers Arch. 451, 35–42.
Staruschenko A., Jeske N.A., Akopian A.N. 2010. Contribution of TRPV1-TRPA1 interaction to the single channel properties of the TRPA1 channel. J. Biol. Chem. 285, 15167–15177.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © D.V. Ilatovskaya, T.S. Pavlov, Y.A. Negulyaev, A. Staruschenko, 2012, published in Biologicheskie Membrany, 2012, Vol. 29, No. 3, pp. 200–208.
The article was translated by the authors.
These authors contributed equally to this work.
Rights and permissions
About this article
Cite this article
Ilatovskaya, D.V., Pavlov, T.S., Negulyaev, Y.A. et al. Regulation of TRPC6 channels by non-steroidal anti-inflammatory drugs. Biochem. Moscow Suppl. Ser. A 6, 265–272 (2012). https://doi.org/10.1134/S1990747812030063
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S1990747812030063