Pathway of Programmed Cell Death and Oxidative Stress Induced by β-Hydroxybutyrate in Dairy Cow Abomasum Smooth Muscle Cells and in Mouse Gastric Smooth Muscle
The administration of exogenous β-hydroxybutyrate (β-HB), as well as fasting and caloric restriction, is a condition associated with β-HB abundance and decreased appetite in animals. Increased β-HB and decreased appetite exist simultaneously in some diseases, such as bovine left displaced abomasums (LDA) and human chronic gastritis. However, the effects of β-HB on stomach injuries have not been explored. To elucidate the possible effects of exogenous β-HB on the stomach, mice were injected intraperitoneally with β-HB, and bovine abomasum smooth muscle cells (BSMCs) were treated with different concentrations of β-HB. We found that β-HB induced BSMCs endoplasmic reticulum- and mitochondria-mediated apoptotic cell death. β-HB promoted Bax expression and caspase-12, -9, and -3 activation while blocking Bcl-2 expression. β-HB also promoted AIF, EndoG release and p53 expression. β-HB acted on key molecules in the apoptotic cell death pathway and increased p38 and c-June NH2-terminal kinase phosphorylation while inhibiting ERK phosphorylation and PCNA expression. β-HB upregulated P27 and P21 mRNA levels while downregulating cyclin and CDK mRNA levels, arresting the cell cycle. These results suggest that BSMCs treated with β-HB can induce oxidative stress, which can be prevented by intracellular calcium chelators BAPTA/AM but not antioxidant NAC. Additionally, these results suggest that β-HB causes ROS generation through a Ca2+-dependent mechanism and that intracellular Ca2+ levels play a critical role in β-HB -induced apoptotic cell death. The impact of β-HB on programmed cell death and oxidative stress in vivo was confirmed in murine experiments. For the first time, we show oxidative stress effects of β-HB on smooth muscle. We propose that β-HB is a possible cause of some stomach diseases, including bovine LDA.
References
[1]
Shimazu T, Hirschey MD, Newman J, He W, Shirakawa K, et al. (2013) Suppression of oxidative stress by beta-hydroxybutyrate, an endogenous histone deacetylase inhibitor. Science 339: 211–214. doi: 10.1126/science.1227166
[2]
Byard RW, Zhou C (2010) Erosive gastritis, Armanni-Ebstein phenomenon and diabetic ketoacidosis. Forensic Sci Med Pathol 6: 304–306. doi: 10.1007/s12024-010-9183-8
[3]
Laeger T, Metges CC, Kuhla B (2010) Role of beta-hydroxybutyric acid in the central regulation of energy balance. Appetite 54: 450–455. doi: 10.1016/j.appet.2010.04.005
[4]
LeBlanc SJ, Leslie KE, Duffield TF (2005) Metabolic predictors of displaced abomasum in dairy cattle. J Dairy Sci 88: 159–170.
[5]
Rehage J, Mertens M, Stockhofe-Zurwieden N, Kaske M, Scholz H (1996) Post surgical convalescence of dairy cows with left abomasal displacement in relation to fatty liver. Schweiz Arch Tierheilkd 138: 361–368.
[6]
Geishauser T, Leslie K, Duffield T, Edge V (1997) Evaluation of aspartate transaminase activity and beta-hydroxybutyrate concentration in blood as tests for prediction of left displaced abomasum in dairy cows. Am J Vet Res 58: 1216–1220.
[7]
Jain SK, Kannan K, Lim G (1998) Ketosis (acetoacetate) can generate oxygen radicals and cause increased lipid peroxidation and growth inhibition in human endothelial cells. Free Radic Biol Med 25: 1083–1088. doi: 10.1016/s0891-5849(98)00140-3
[8]
Laffel L (1999) Ketone bodies: a review of physiology, pathophysiology and application of monitoring to diabetes. Diabetes Metab Res Rev 15: 412–426. doi: 10.1002/(sici)1520-7560(199911/12)15:6<412::aid-dmrr72>3.0.co;2-8
[9]
Fulop M, Murthy V, Michilli A, Nalamati J, Qian Q, et al. (1999) Serum beta-hydroxybutyrate measurement in patients with uncontrolled diabetes mellitus. Arch Intern Med 159: 381–384. doi: 10.1001/archinte.159.4.381
[10]
Rossi R, Dorig S, Del Prete E, Scharrer E (2000) Suppression of feed intake after parenteral administration of D-beta-hydroxybutyrate in pygmy goats. J Vet Med A Physiol Pathol Clin Med 47: 9–16. doi: 10.1046/j.1439-0442.2000.00255.x
[11]
Cahill GF Jr, Veech RL (2003) Ketoacids? Good medicine? Trans Am Clin Climatol Assoc 114: 149–161 discussion 162–143.
[12]
Taggart AK, Kero J, Gan X, Cai TQ, Cheng K, et al. (2005) (D)-beta-Hydroxybutyrate inhibits adipocyte lipolysis via the nicotinic acid receptor PUMA-G. J Biol Chem 280: 26649–26652. doi: 10.1074/jbc.c500213200
[13]
Cheng B, Lu H, Bai B, Chen J (2013) d-beta-Hydroxybutyrate inhibited the apoptosis of PC12 cells induced by H2O2 via inhibiting oxidative stress. Neurochem Int 62: 620–625. doi: 10.1016/j.neuint.2012.09.011
[14]
Pelletier A, Coderre L (2007) Ketone bodies alter dinitrophenol-induced glucose uptake through AMPK inhibition and oxidative stress generation in adult cardiomyocytes. Am J Physiol Endocrinol Metab 292: E1325–1332. doi: 10.1152/ajpendo.00186.2006
[15]
Guh JY, Chuang TD, Chen HC, Hung WC, Lai YH, et al. (2003) Beta-hydroxybutyrate-induced growth inhibition and collagen production in HK-2 cells are dependent on TGF-beta and Smad3. Kidney Int 64: 2041–2051. doi: 10.1046/j.1523-1755.2003.00330.x
[16]
Skinner R, Trujillo A, Ma X, Beierle EA (2009) Ketone bodies inhibit the viability of human neuroblastoma cells. J Pediatr Surg 44: 212–216 discussion 216. doi: 10.1016/j.jpedsurg.2008.10.042
[17]
Lin SY, Lai WW, Ho CC, Yu FS, Chen GW, et al. (2009) Emodin induces apoptosis of human tongue squamous cancer SCC-4 cells through reactive oxygen species and mitochondria-dependent pathways. Anticancer Res 29: 327–335.
[18]
Ho SY, Chen WC, Chiu HW, Lai CS, Guo HR, et al. (2009) Combination treatment with arsenic trioxide and irradiation enhances apoptotic effects in U937 cells through increased mitotic arrest and ROS generation. Chem Biol Interact 179: 304–313. doi: 10.1016/j.cbi.2008.12.015
[19]
Turi S, Nemeth I, Torkos A, Saghy L, Varga I, et al. (1997) Oxidative stress and antioxidant defense mechanism in glomerular diseases. Free Radic Biol Med 22: 161–168. doi: 10.1016/s0891-5849(96)00284-5
[20]
Ip SW, Chu YL, Yu CS, Chen PY, Ho HC, et al. (2012) Bee venom induces apoptosis through intracellular Ca2+ -modulated intrinsic death pathway in human bladder cancer cells. Int J Urol 19: 61–70. doi: 10.1111/j.1442-2042.2011.02876.x
[21]
Son YO, Lee JC, Hitron JA, Pan J, Zhang Z, et al. (2010) Cadmium induces intracellular Ca2+- and H2O2-dependent apoptosis through JNK- and p53-mediated pathways in skin epidermal cell line. Toxicol Sci 113: 127–137. doi: 10.1093/toxsci/kfp259
[22]
Li J, Lee B, Lee AS (2006) Endoplasmic reticulum stress-induced apoptosis: multiple pathways and activation of p53-up-regulated modulator of apoptosis (PUMA) and NOXA by p53. J Biol Chem 281: 7260–7270. doi: 10.1074/jbc.m509868200
[23]
Jacobson J, Duchen MR (2002) Mitochondrial oxidative stress and cell death in astrocytes–requirement for stored Ca2+ and sustained opening of the permeability transition pore. J Cell Sci 115: 1175–1188.
[24]
Camello-Almaraz C, Gomez-Pinilla PJ, Pozo MJ, Camello PJ (2006) Mitochondrial reactive oxygen species and Ca2+ signaling. Am J Physiol Cell Physiol 291: C1082–1088. doi: 10.1152/ajpcell.00217.2006
[25]
Brookes P, Darley-Usmar VM (2002) Hypothesis: the mitochondrial NO(*) signaling pathway, and the transduction of nitrosative to oxidative cell signals: an alternative function for cytochrome C oxidase. Free Radic Biol Med 32: 370–374. doi: 10.1016/s0891-5849(01)00805-x
[26]
Distelhorst CW, Dubyak G (1998) Role of calcium in glucocorticosteroid-induced apoptosis of thymocytes and lymphoma cells: resurrection of old theories by new findings. Blood 91: 731–734.
[27]
Yan Y, Wei CL, Zhang WR, Cheng HP, Liu J (2006) Cross-talk between calcium and reactive oxygen species signaling. Acta Pharmacol Sin 27: 821–826. doi: 10.1111/j.1745-7254.2006.00390.x
Chang L, Karin M (2001) Mammalian MAP kinase signalling cascades. Nature 410: 37–40.
[30]
el-Deiry WS, Kern SE, Pietenpol JA, Kinzler KW, Vogelstein B (1992) Definition of a consensus binding site for p53. Nat Genet 1: 45–49. doi: 10.1038/ng0492-45
[31]
Schuler M, Green DR (2001) Mechanisms of p53-dependent apoptosis. Biochem Soc Trans 29: 684–688. doi: 10.1042/bst0290684
[32]
Chipuk JE, Kuwana T, Bouchier-Hayes L, Droin NM, Newmeyer DD, et al. (2004) Direct activation of Bax by p53 mediates mitochondrial membrane permeabilization and apoptosis. Science 303: 1010–1014. doi: 10.1126/science.1092734
[33]
Marchenko ND, Zaika A, Moll UM (2000) Death signal-induced localization of p53 protein to mitochondria. A potential role in apoptotic signaling. J Biol Chem 275: 16202–16212. doi: 10.1074/jbc.275.21.16202
[34]
Wang Y, Solt LA, Kojetin DJ, Burris TP (2012) Regulation of p53 stability and apoptosis by a ROR agonist. PLoS One 7: e34921. doi: 10.1371/journal.pone.0034921
[35]
Kim R, Emi M, Tanabe K (2006) Role of mitochondria as the gardens of cell death. Cancer Chemother Pharmacol 57: 545–553. doi: 10.1007/s00280-005-0111-7
[36]
Scorrano L, Oakes SA, Opferman JT, Cheng EH, Sorcinelli MD, et al. (2003) BAX and BAK regulation of endoplasmic reticulum Ca2+: a control point for apoptosis. Science 300: 135–139. doi: 10.1126/science.1081208
[37]
Acehan D, Jiang X, Morgan DG, Heuser JE, Wang X, et al. (2002) Three-dimensional structure of the apoptosome: implications for assembly, procaspase-9 binding, and activation. Mol Cell 9: 423–432. doi: 10.1016/s1097-2765(02)00442-2
[38]
Nutt LK, Chandra J, Pataer A, Fang B, Roth JA, et al. (2002) Bax-mediated Ca2+ mobilization promotes cytochrome c release during apoptosis. J Biol Chem 277: 20301–20308. doi: 10.1074/jbc.m201604200
[39]
Jacotot E, Costantini P, Laboureau E, Zamzami N, Susin SA, et al. (1999) Mitochondrial membrane permeabilization during the apoptotic process. Ann N Y Acad Sci 887: 18–30. doi: 10.1111/j.1749-6632.1999.tb07919.x
[40]
Jayanthi S, Deng X, Noailles PA, Ladenheim B, Cadet JL (2004) Methamphetamine induces neuronal apoptosis via cross-talks between endoplasmic reticulum and mitochondria-dependent death cascades. FASEB J 18: 238–251. doi: 10.1096/fj.03-0295com
[41]
Saelens X, Festjens N, Vande Walle L, van Gurp M, van Loo G, et al. (2004) Toxic proteins released from mitochondria in cell death. Oncogene 23: 2861–2874. doi: 10.1038/sj.onc.1207523
[42]
Li P, Nijhawan D, Budihardjo I, Srinivasula SM, Ahmad M, et al. (1997) Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 91: 479–489. doi: 10.1016/s0092-8674(00)80434-1
[43]
Earnshaw WC, Martins LM, Kaufmann SH (1999) Mammalian caspases: structure, activation, substrates, and functions during apoptosis. Annu Rev Biochem 68: 383–424. doi: 10.1146/annurev.biochem.68.1.383
[44]
Morishima N, Nakanishi K, Takenouchi H, Shibata T, Yasuhiko Y (2002) An endoplasmic reticulum stress-specific caspase cascade in apoptosis. Cytochrome c-independent activation of caspase-9 by caspase-12. J Biol Chem 277: 34287–34294. doi: 10.1074/jbc.m204973200
[45]
Tantral L, Malathi K, Kohyama S, Silane M, Berenstein A, et al. (2004) Intracellular calcium release is required for caspase-3 and -9 activation. Cell Biochem Funct 22: 35–40. doi: 10.1002/cbf.1050
[46]
Liu ZM, Chen GG, Vlantis AC, Tse GM, Shum CK, et al. (2007) Calcium-mediated activation of PI3K and p53 leads to apoptosis in thyroid carcinoma cells. Cell Mol Life Sci 64: 1428–1436. doi: 10.1007/s00018-007-7107-x
[47]
Maga G, Hubscher U (2003) Proliferating cell nuclear antigen (PCNA): a dancer with many partners. J Cell Sci 116: 3051–3060. doi: 10.1242/jcs.00653
[48]
Miyachi K, Fritzler MJ, Tan EM (1978) Autoantibody to a nuclear antigen in proliferating cells. J Immunol 121: 2228–2234.
