Abstract
Retinol-binding protein 4 (RBP4) protein is a kind of adipokines synthesized and secreted by the liver, which has been verified to play important roles in liver metabolism and energy homeostasis. However, the effects of RBP4 on hepatic lipid accumulation are still elusive in fish. In the present study, we cloned and characterized the RBP4 gene in Schizothorax prenanti (S. prenanti). RBP4 gene was specifically expressed in the liver and abdominal adipose tissue. Palmitic acid (PA; 400 μM) can significantly increase lipid deposition in primary hepatocytes after 12 h of treatment. Furthermore, RBP4 knockdown can relieve the excessive lipid deposition and endoplasmic reticulum stress in the hepatocytes caused by PA. The inhibition of RBP4 abolished the ability of PA to induce the expression of genes involved in lipogenesis and endoplasmic reticulum stress. These results demonstrate that RBP4 inhibition attenuated PA-induced lipid deposition and endoplasmic reticulum stress in hepatocytes of S. prenanti. This study could contribute to improve the understanding of RBP4 functions in the PA-induced lipid deposition in hepatocytes of fish.
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Abbreviations
- RBP4:
-
Retinol-binding protein 4
- FAS:
-
Fatty acid synthase
- ACC:
-
Acetyl-CoA carboxylase
- SCD1:
-
Stearoyl-Coenzyme A desaturase 1
- L-FABP:
-
Liver-basic fatty acid binding protein b
- GK:
-
Glucokinase
- PK:
-
Pyruvate kinase
- PEPCK:
-
Phosphoenolpyruvate carboxykinase
- G6Pase:
-
Glucose-6-phosphatase
- FBPase:
-
Fructose-1,6-bisphosphatase
- XBP1s:
-
Spliced X-box binding protein 1
- GRP78:
-
Glucose-regulated protein 78
- CHOP:
-
DNA damage inducible transcript 3
References
Cha S-H, Hwang Y, Kim K-N, Jun H-S (2018) Palmitate induces nitric oxide production and inflammatory cytokine expression in zebrafish. Fish Shellfish Immunol 79:163–167. https://doi.org/10.1016/j.fsi.2018.05.025
Chen J, Wu HQ, Shi YH, Li CH, Li MY (2009) The effects of environmental salinity on trunk kidney proteome of juvenile ayu (Plecoglossus altivelis). Comp Biochem Physiol D Genomics Proteomics 4(4):263–267. https://doi.org/10.1016/j.cbd.2009.06.003
Chen G, Wu K, Zhao T, Ling S, Liu W, Luo Z (2020) miR-144 mediates high fat-induced changes of cholesterol metabolism via direct regulation of C/EBPalpha in the liver and isolated hepatocytes of yellow catfish. J Nutr 150(3):464–474. https://doi.org/10.1093/jn/nxz282
Chiefari E, Paonessa F, Iiritano S, Le Pera I, Palmieri D, Brunetti G, Lupo A, Colantuoni V, Foti D, Gulletta E, De Sarro G, Fusco A, Brunetti A (2009) The cAMP-HMGA1-RBP4 system: a novel biochemical pathway for modulating glucose homeostasis. BMC Biol 7:24. https://doi.org/10.1186/1741-7007-7-24
Ding Q, Zhang Z, Ran C, He S, Yang Y, Du Z, Zhang J, Zhou Z (2018) The hepatotoxicity of palmitic acid in zebrafish involves the intestinal microbiota. J Nutr 148(8):1217–1228. https://doi.org/10.1093/jn/nxy084
Eguchi A, Iwasa M (2021) The role of elevated liver-type fatty acid-binding proteins in liver diseases. Pharm Res 38(1):89–95. https://doi.org/10.1007/s11095-021-02998-x
Fedders R, Muenzner M, Weber P, Sommerfeld M, Knauer M, Kedziora S, Kast N, Heidenreich S, Raila J, Weger S, Henze A, Schupp M (2018) Liver-secreted RBP4 does not impair glucose homeostasis in mice. J Biol Chem 293(39):15269–15276. https://doi.org/10.1074/jbc.RA118.004294
Flower DR, North ACT, Sansom CE (2000) The lipocalin protein family: structural and sequence overview. Biochim Biophys Acta Protein Struct Mol Enzymol 1482(1):9–24. https://doi.org/10.1016/S0167-4838(00)00148-5
Friedman SL, Neuschwander-Tetri BA, Rinella M, Sanyal AJ (2018) Mechanisms of NAFLD development and therapeutic strategies. Nat Med 24(7):908–922. https://doi.org/10.1038/s41591-018-0104-9
Goodman SD (1980) Plasma retinol-binding protein. N Y Acad Sci 348(none):378–390
Larbi Ayisi C, Zhao J, Wu JW (2018) Replacement of fish oil with palm oil: effects on growth performance, innate immune response, antioxidant capacity and disease resistance in Nile tilapia (Oreochromis niloticus). PLoS ONE 13(4):e0196100. https://doi.org/10.1371/journal.pone.0196100
Lee SA, Yuen JJ, Jiang H, Kahn BB, Blaner WS (2016) Adipocyte-specific overexpression of retinol-binding protein 4 causes hepatic steatosis in mice. Hepatology 64(5):1534–1546. https://doi.org/10.1002/hep.28659
Li X, Ji R, Cui K, Chen Q, Chen Q, Fang W, Mai K, Zhang Y, Xu W, Ai Q (2019) High percentage of dietary palm oil suppressed growth and antioxidant capacity and induced the inflammation by activation of TLR-NF-kappaB signaling pathway in large yellow croaker (Larimichthys crocea). Fish Shellfish Immunol 87:600–608. https://doi.org/10.1016/j.fsi.2019.01.055
Li J, Fan Q, Cai H, Deng J, Ming F, Li J, Zeng M, Ma M, Zhao P, Liang Q, Jia J, Zhang S, Zhang L (2020) Identification of RBP4 from bighead carp (Hypophthalmichthys nobilis) / silver carp (Hypophthalmichthys molitrix) and effects of CpG ODN on RBP4 expression under A. hydrophila challenge. Fish Shellfish Immunol 100:476–488. https://doi.org/10.1016/j.fsi.2020.03.036
Ma X, Zhou Z, Chen Y, Wu Y, Liu Y (2016) RBP4 functions as a hepatokine in the regulation of glucose metabolism by the circadian clock in mice. Diabetologia 59(2):354–362. https://doi.org/10.1007/s00125-015-3807-1
Park K-H, Ye Z-W, Zhang J, Kim S-H (2019) Palmitic acid-enriched diet induces hepatic steatosis and injury in adult zebrafish. Zebrafish 16(6):497–504. https://doi.org/10.1089/zeb.2019.1758
Raghu P, Sivakumar B (2004) Interactions amongst plasma retinol-binding protein, transthyretin and their ligands: implications in vitamin A homeostasis and transthyretin amyloidosis. Biochim Biophys Acta Proteins Proteomics 1703(1):1–9. https://doi.org/10.1016/j.bbapap.2004.09.023
Salmerón C (2018) Adipogenesis in fish. J Exp Biol 221(Suppl 1). https://doi.org/10.1242/jeb.161588
Sanyal A (2019) Past, present and future perspectives in nonalcoholic fatty liver disease. Nat Rev Gastroenterol Hepatol 16:1. https://doi.org/10.1038/s41575-019-0144-8
Stefan N, Hennige AM, Staiger H, Machann J, Schick F, Schleicher E, Fritsche A, Haring HU (2007) High circulating retinol-binding protein 4 is associated with elevated liver fat but not with total, subcutaneous, visceral, or intramyocellular fat in humans. Diabetes Care 30(5):1173–1178. https://doi.org/10.2337/dc06-2342
Thompson SJ, Sargsyan A, Lee SA, Yuen JJ, Cai J, Smalling R (2017) Hepatocytes are the principal source of circulating RBP4 in mice. Diabetes 66(1):58–63. https://doi.org/10.2337/db16-0286/-/DC1
Tingaud-Sequeira A, Forgue J, Andre M, Babin PJ (2006) Epidermal transient down-regulation of retinol-binding protein 4 and mirror expression of apolipoprotein Eb and estrogen receptor 2a during zebrafish fin and scale development. Dev Dyn 235(11):3071–3079. https://doi.org/10.1002/dvdy.20921
Wan Ahmad WA, Stone DA, Schuller KA (2013) Dietary fish oil replacement with palm or poultry oil increases fillet oxidative stability and decreases liver glutathione peroxidase activity in barramundi (Lates calcarifer). Fish Physiol Biochem 39(6):1631–1640. https://doi.org/10.1007/s10695-013-9815-5
Wang L, Zhu P, Mo Q, Luo W, Du Z, Jiang J, Yang S, Zhao L, Gong Q, Wang Y (2021) Comprehensive analysis of full-length transcriptomes of Schizothorax prenanti by single-molecule long-read sequencing. Genomics. https://doi.org/10.1016/j.ygeno.2021.01.009
Wei Y, Wang D, Topczewski F, Pagliassotti MJ (2006) Saturated fatty acids induce endoplasmic reticulum stress and apoptosis independently of ceramide in liver cells. Am J Physiol Endocrinol Metab 291(2):E275-281. https://doi.org/10.1152/ajpendo.00644.2005
Wei Y, Wang D, Pagliassotti MJ (2007) Saturated fatty acid-mediated endoplasmic reticulum stress and apoptosis are augmented by trans-10, cis-12-conjugated linoleic acid in liver cells. Mol Cell Biochem 303(1):105–113. https://doi.org/10.1007/s11010-007-9461-2
Weil C, Lefèvre F, Bugeon J (2012) Characteristics and metabolism of different adipose tissues in fish. Rev Fish Biol Fish 23(2):157–173. https://doi.org/10.1007/s11160-012-9288-0
Wu, Wei, Yang, Zhao, Luo (2019) Effects of fat and fatty acids on the formation of autolysosomes in the livers from yellow catfish Pelteobagrus fulvidraco. Genes 10(10). https://doi.org/10.3390/genes10100751
Xia M, Liu Y, Guo H, Wang D, Wang Y, Ling W (2013) Retinol binding protein 4 stimulates hepatic sterol regulatory element-binding protein 1 and increases lipogenesis through the peroxisome proliferator-activated receptor-gamma coactivator 1beta-dependent pathway. Hepatology 58(2):564–575. https://doi.org/10.1002/hep.26227
Xiang X, Han S, Xu D, Chen Q, Ji R, Zhao Z, Du J, Mai K, Ai Q (2021) Oleic and palmitic acids induce hepatic angiopoietin-like 4 expression predominantly via PPAR-gamma in Larimichthys crocea. Br J Nutr:1–27. https://doi.org/10.1017/S000711452100386X
Xu H, Dong X, Zuo R, Mai K, Ai Q (2016) Response of juvenile Japanese seabass (Lateolabrax japonicus) to different dietary fatty acid profiles: growth performance, tissue lipid accumulation, liver histology and flesh texture. Aquaculture 461:40–47. https://doi.org/10.1016/j.aquaculture.2016.04.023
Yang Q, Graham TE, Mody N, Preitner F, Peroni OD, Zabolotny JM, Kotani K, Quadro L, Kahn BB (2005) Serum retinol binding protein 4 contributes to insulin resistance in obesity and type 2 diabetes. Nature 436(7049):356–362. https://doi.org/10.1038/nature03711
Yang SS, Yu CB, Luo Z, Luo WL, Zhang J, Xu JX, Xu WN (2019) Berberine attenuates sodium palmitate-induced lipid accumulation, oxidative stress and apoptosis in grass carp (Ctenopharyngodon idella) hepatocyte in vitro. Fish Shellfish Immunol 88:518–527. https://doi.org/10.1016/j.fsi.2019.02.055
Yang B, Ji R, Li X, Fang W, Chen Q, Chen Q, Xu W, Mai K, Ai Q (2021) Activation of autophagy relieves linoleic acid-induced inflammation in large yellow croaker (Larimichthys crocea). Front Immunol 12:649385. https://doi.org/10.3389/fimmu.2021.649385
Zhang J, Liu Q, Pang Y, Xu X, Cui K, Zhang Y, Mai K, Ai Q (2020) Molecular cloning and the involvement of IRE1alpha-XBP1s signaling pathway in palmitic acid induced - inflammation in primary hepatocytes from large yellow croaker (Larimichthys crocea). Fish Shellfish Immunol 98:112–121. https://doi.org/10.1016/j.fsi.2019.12.089
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This study was supported by the grant from the National Natural Science Foundation of China (31602148).
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Data curation, funding acquisition, and project administration: Yan Wang; Writing-original draft preparation and data validation: Peng Zhu; Methodology and writing-reviewing and editing: Jiahui Ni and Qilang Mo; Software: Wei Luo and Zongjun Du; Resources: Jun Jiang; Formal analysis: Song Yang and Liulan Zhao; Supervision and investigation: Quan Gong. All authors read and approved the final manuscript.
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Wang, Y., Zhu, P., Ni, J. et al. Molecular and functional characterization of the retinol-binding protein 4 (RBP4) in hepatocytes of Schizothorax prenanti in response to palmitic acid. Fish Physiol Biochem 48, 449–459 (2022). https://doi.org/10.1007/s10695-022-01060-w
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DOI: https://doi.org/10.1007/s10695-022-01060-w