Inhibition of mitochondrial phosphate carrier prevents high phosphate-induced superoxide generation and vascular calcification
2
조회
0
다운로드
0
인용
0
참고문헌
초록
Vascular calcification is a serious complication of hyperphosphatemia that causes cardiovascular morbidity and mortality. Previous studies have reported that plasmalemmal phosphate (Pi) transporters, such as PiT-1/2, mediate depolarization, Ca2+ influx, oxidative stress, and calcific changes in vascular smooth muscle cells (VSMCs). However, the pathogenic mechanism of mitochondrial Pi uptake in vascular calcification associated with hyperphosphatemia has not been elucidated. We demonstrated that the phosphate carrier (PiC) is the dominant mitochondrial Pi transporter responsible for high Pi-induced superoxide generation, osteogenic gene upregulation, and calcific changes in primary VSMCs isolated from rat aortas. Notably, acute incubation with high Pi markedly increased the protein abundance of PiC via ERK1/2- and mTOR-dependent translational upregulation. Genetic suppression of PiC prevented Pi-induced ERK1/2 activation, superoxide production, osteogenic differentiation, and vascular calcification of VSMCs in vitro and aortic rings ex vivo. Pharmacological inhibition of mitochondrial Pi transport using butyl malonate (BMA) or mersalyl abolished all pathologic changes involved in high Pi-induced vascular calcification. BMA or mersalyl also effectively prevented osteogenic gene upregulation and calcification of aortas from 5/6 subtotal nephrectomized mice fed a high-Pi diet. Our results suggest that mitochondrial Pi uptake via PiC is a critical molecular mechanism mediating mitochondrial superoxide generation and pathogenic calcific changes, which could be a novel therapeutic target for treating vascular calcification associated with hyperphosphatemia.
Abstract
Vascular calcification is a serious complication of hyperphosphatemia that causes cardiovascular morbidity and mortality. Previous studies have reported that plasmalemmal phosphate (Pi) transporters, such as PiT-1/2, mediate depolarization, Ca2+ influx, oxidative stress, and calcific changes in vascular smooth muscle cells (VSMCs). However, the pathogenic mechanism of mitochondrial Pi uptake in vascular calcification associated with hyperphosphatemia has not been elucidated. We demonstrated that the phosphate carrier (PiC) is the dominant mitochondrial Pi transporter responsible for high Pi-induced superoxide generation, osteogenic gene upregulation, and calcific changes in primary VSMCs isolated from rat aortas. Notably, acute incubation with high Pi markedly increased the protein abundance of PiC via ERK1/2- and mTOR-dependent translational upregulation. Genetic suppression of PiC prevented Pi-induced ERK1/2 activation, superoxide production, osteogenic differentiation, and vascular calcification of VSMCs in vitro and aortic rings ex vivo. Pharmacological inhibition of mitochondrial Pi transport using butyl malonate (BMA) or mersalyl abolished all pathologic changes involved in high Pi-induced vascular calcification. BMA or mersalyl also effectively prevented osteogenic gene upregulation and calcification of aortas from 5/6 subtotal nephrectomized mice fed a high-Pi diet. Our results suggest that mitochondrial Pi uptake via PiC is a critical molecular mechanism mediating mitochondrial superoxide generation and pathogenic calcific changes, which could be a novel therapeutic target for treating vascular calcification associated with hyperphosphatemia.
참고문헌 (37)
- Vervloet, M. G, "Prevention and treatment of hyperphosphatemia in chronic kidney disease", Kidney Int, 93, pp.1060~1072 (2018)
- Komaba, H., "Phosphate-a poison for humans?", Kidney Int, 90, pp.753~763 (2016)
- Kuro-o, M., "Overview of the FGF23-Klotho axis", Pediatr. Nephrol, 25, pp.583~590 (2010)
- Moore, L. W., "Association of dietary phosphate and serum phosphorus concentration by levels of kidney function", Am. J. Clin. Nutr, 102, pp.444~453 (2015)
- Lanzer, P., "Medial vascular calcification revisited : review and perspectives", Eur. Heart J, 35, pp.1515~1525 (2014)
- Thongprayoon, C., "Impact of serum phosphate changes on in-hospital mortality", BMC Nephrol, 21, pp.427~ (2020)
- Shanahan, C. M., "Arterial calcification in chronic kidney disease : key roles for calcium and phosphate", Circ. Res, 109, pp.697~711 (2011)
- Durham, A. L., "Role of smooth muscle cells in vascular calcification : implications in atherosclerosis and arterial stiffness", Cardiovasc. Res, 114, pp.590~600 (2018)
- Proudfoot, D., "Apoptosis regulates human vascular calcification in vitro : evidence for initiation of vascular calcification by apoptotic bodies", Circ. Res, 87, pp.1055~1062 (2000)
- Rahabi-Layachi, H., "Distinct effects of inorganic phosphate on cell cycle and apoptosis in human vascular smooth muscle cells", J. Cell Physiol, 230, pp.347~355 (2015)
- Nguyen, N. T., "Oxidative stress by Ca(2+)overload is critical for phosphateinduced vascular calcification", Am. J. Physiol. Heart Circ. Physiol, 319, pp.H1302~h1312 (2020)
- Nguyen, T. T., "Intracellular alkalinization by phosphate uptake via type III sodium-phosphate cotransporter participates in high-phosphate-induced mitochondrial oxidative stress and defective insulin secretion", FASEB J, 30, pp.3979~3988 (2016)
- Zhao, M. M., "Mitochondrial reactive oxygen species promote p65 nuclear translocation mediating high-phosphate-induced vascular calcification in vitro and in vivo", Kidney Int, 79, pp.1071~1079 (2011)
- Seifert, E. L., "The mitochondrial phosphate carrier : role in oxidative metabolism, calcium handling and mitochondrial disease", Biochem. Biophys. Res. Commun, 464, pp.369~375 (2015)
- Johnson, R. N., "The transport of inorganic phosphate by the mitochondrial dicarboxylate carrier", Biochem. J, 134, pp.769~774 (1973)
- Fiermonte, G., "Identification of the mitochondrial ATP-Mg", Pi transporter. Bacterial expression, reconstitution, functional characterization, and tissue distribution, 2004, pp.279, 30722~30730
- Vozza, A., "UCP2 transports C4 metabolites out of mitochondria, regulating glucose and glutamine oxidation", Proc. Natl Acad. Sci. USA, 111, pp.960~965 (2014)
- Dolce, V., "Tissue-specific expression of the two isoforms of the mitochondrial phosphate carrier in bovine tissues", FEBS Lett, 399, pp.95~98 (1996)
- DAS, K., "Predominant expression of the mitochondrial dicarboxylate carrier in white adipose tissue", Biochem. J, 344, pp.313~320 (1999)
- Palmieri, F., "The mitochondrial transporter family SLC25 : identification, properties and physiopathology", Mol. Asp. Med, 34, pp.465~484 (2013)
- Hoffman, N. E., "SLC25A23 augments mitochondrial Ca(2)(+)uptake, interacts with MCU, and induces oxidative stress-mediated cell death", Mol. Biol. Cell, 25, pp.936~947 (2014)
- Patel, J. J., "Isolation, culture, and characterization of vascular smooth muscle cells", Methods Mol. Biol, 1430, pp.91~105 (2016)
- Tan, R. Z., "An optimized 5", 6 nephrectomy mouse model based on unilateral kidney ligation and its application in renal fibrosis research, 2019, pp.41, 555~566
- Schneider-Poetsch, T., "Inhibition of eukaryotic translation elongation by cycloheximide and lactimidomycin", Nat. Chem. Biol, 6, pp.209~217 (2010)
- Kisselev, A. F., "Proteasome inhibitors : from research tools to drug candidates", Chem. Biol, 8, pp.739~758 (2001)
- Nguyen, T. T., "Mitochondrial oxidative stress mediates high-phosphateinduced secretory defects and apoptosis in insulin-secreting cells", Am. J. Physiol. Endocrinol. Metab, 308, pp.E933~E941 (2015)
- Quan, X., "Mitochondrial phosphate transport during nutrient stimulation of INS-1E insulinoma cells", Mol. Cell. Endocrinol, 381, pp.198~209 (2013)
- Kwong, J. Q., "Genetic deletion of the mitochondrial phosphate carrier desensitizes the mitochondrial permeability transition pore and causes cardiomyopathy", Cell Death Differ, 21, pp.1209~1217 (2014)
- Seifert, E. L., "Natural and induced mitochondrial phosphate carrier loss : differential dependence of mitochondrial metabolism and dynamics and cell survival on the extent of depletion", J. Biol. Chem, 291, pp.26126~26137 (2016)
- Varanyuwatana, P., "The roles of phosphate and the phosphate carrier in the mitochondrial permeability transition pore", Mitochondrion, 12, pp.120~125 (2012)
- Zhao, G., "Activation of nuclear factor-kappa B accelerates vascular calcification by inhibiting ankylosis protein homolog expression", Kidney Int, 82, pp.34~44 (2012)
- Ruprecht, J. J., "The SLC25 mitochondrial carrier family: structure and mechanism", Trends Biochem. Sci, 45, pp.244~258 (2020)
- Nguyen, N. T., "Oxidative stress related to plasmalemmal and mitochondrial phosphate transporters in vascular calcification", Antioxidants, 11, pp.494~ (2022)
- Kuro, O. M., "Phosphate and Klotho", Kidney Int, 79121, pp.S20~S23 (2011)
- Nakano, C., "Combined use of vitamin D status and FGF23 for risk stratification of renal outcome", Clin. J. Am. Soc. Nephrol, 7, pp.810~819 (2012)
- Shroff, R., "Mechanistic Insights into Vascular Calcification in CKD", J. Am. Soc. Nephrol, 24, pp.179~189 (2013)
- Huang, M., "Oxidative stress contributes to vascular calcification in patients with chronic kidney disease", J. Mol. Cell. Cardiol, 138, pp.256~268 (2020)