Animal Bioscience

  • 제38권12호
  • /
  • Pages.2558-2567
  • /
  • 2025
  • /
  • 2765-0189(pISSN)
  • /
  • 2765-0235(eISSN)
아세아태평양축산학회 (Asian Australasian Association of Animal Production Societies)
권호 표지
DOI QR코드

DOI QR Code

Effects of the DUSP6 gene on the proliferation and differentiation of porcine subcutaneous preadipocytes

  • Xiangxiang Yang (College of Animal Science and Technology, Anhui Agricultural University) ;
  • Xiaohan Sun (College of Animal Science and Technology, Anhui Agricultural University) ;
  • Zimeng Du (College of Animal Science and Technology, Anhui Agricultural University) ;
  • Jundong Yi (College of Animal Science and Technology, Anhui Agricultural University) ;
  • Qiuyan Wang (College of Animal Science and Technology, Anhui Agricultural University) ;
  • Ding Yin (College of Animal Science and Technology, Anhui Agricultural University) ;
  • Yalan Zhang (College of Animal Science and Technology, Anhui Agricultural University) ;
  • Xiaoling Ding (College of Animal Science and Technology, Anhui Agricultural University) ;
  • Xianrui Zheng (College of Animal Science and Technology, Anhui Agricultural University) ;
  • Xiaodong Zhang (College of Animal Science and Technology, Anhui Agricultural University) ;
  • Zongjun Yin (College of Animal Science and Technology, Anhui Agricultural University) ;
  • Yueyun Ding (College of Animal Science and Technology, Anhui Agricultural University)
  • 투고 : 2025.03.16
  • 심사 : 2025.06.17
  • 발행 : 2025.12.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Objective: Dual-specificity protein phosphatase 6 (DUSP6), also known as mitogen-activated protein kinase phosphatase 3 (MKP-3), was considered as a functional candidate gene for white fat accumulation in mice. However, the physiological function of the DUSP6 gene on white adipocyte adipogenesis in farm animals remains unknown. In this study, we aimed to clarify the effect of DUSP6 on porcine subcutaneous preadipocyte proliferation and differentiation. Methods: We first make clear that the patterns of DUSP6 expression is associated with fat contents in porcine fat deposition related tissues. Porcine subcutaneous preadipocytes were isolated and induced to differentiation. Small interfering RNAs were applied to deplete DUSP6. MTT assay, CCK-8 analysis, Oil Red O staining, triglyceride determination and reverse transcription quantitative polymerase chain reaction were applied to study the regulatory role of DUSP6 during adipocyte adipogenesis in pigs. Results: We found that the expression levels of DUSP6 were significantly higher in backfat and longissimus dorsi tissues from fat-type pigs than in those from lean-type pigs. Consistently, the significantly induced expression of DUSP6 was also observed in differentiated adipocytes. In addition, knockdown of DUSP6 greatly inhibited preadipocytes proliferation, through the decreased cell viability and downregulated mRNA expressions of cell proliferation-associated genes, including PCNA, CDK1, CDK2. Furthermore, knockdown of DUSP6 significantly inhibited preadipocytes differentiation, as evidenced by markedly reduced lipid droplet formation, attenuated triglyceride accumulation and downregulated expression levels of adipogenic transcription masters (PPARγ, C/EBPβ, FASN and FABP4) in DUSP6 knockdown cells. Conclusion: Our results demonstrate that DUSP6 is required for white adipocyte adipogenesis in pigs.

키워드

과제정보

This research was funded by the National Key Research and Development Program of China (2021YFD1301200), the Joint Research Project on Local Pig Breeding in Anhui Province (340000211260001000431), the National Natural Science Foundation of China (32573167), the Collaborative Innovation Project of Universities in Anhui Province (GXXT-2023-059), and the Anhui Provincial Modern Industrial Technology System of Swine (AHCYJSTX-05-03, AHCYJSTX-05-01).

참고문헌

  1. Lingvay I, Cohen RV, le Roux CW, Sumithran P. Obesity in adults. Lancet 2024;404:972-87. https://doi.org/10.1016/S0140-6736(24)01210-8
  2. Zhang Y, Feng G, Zhang W, Liu X. Natural compounds exert anti-obesity effects by regulating cytokines. Phytother Res 2025;39:2824-47. https://doi.org/10.1002/ptr.8508
  3. Chilamakuri SN, Manasa N, Thalla M, et al. Role of microneedles for improved treatment of obesity: progress and challenges. Mol Pharm 2025;22:2350-71. https://doi.org/10.1021/acs.Molpharmaceut.4c01115
  4. Welsh A, Hammad M, Piña IL, Kulinski J. Obesity and cardiovascular health. Eur J Prev Cardiol 2024;31:1026-35. https://doi.org/10.1093/eurjpc/zwae025
  5. Pahlavani M, Pham K, Kalupahana NS, et al. Thermogenic adipose tissues: promising therapeutic targets for metabolic diseases. J Nutr Biochem 2025;137:109832. https://doi.org/10.1016/j.jnutbio.2024.109832
  6. Kazemi N, Ramazani E, Tayarani-Najaran Z. In vitro effects of phytochemicals on adipogenesis with a focus on molecular mechanisms: a systematic review. Iran J Basic Med Sci 2025; 28:401-17. https://doi.org/10.22038/IJBMS.2025.78924.17090
  7. Ma Y, Han L, Hou S, et al. Fatty acids and volatile flavor components of adipose tissue from local Tibetan sheep in Qinghai with dietary supplementation of Palm Kernel Meal (PKM). Animals 2024;14:2113. https://doi.org/10.3390/ani14142113
  8. Zhang S, Huang Y, Zheng C, et al. Leucine improves the growth performance, carcass traits, and lipid nutritional quality of pork in Shaziling pigs. Meat Sci 2024;210:109435. https://doi.org/10.1016/j.meatsci.2024.109435
  9. Liu S, Yang Y, Luo H, Pang W, Martin GB. Fat deposition and partitioning for meat production in cattle and sheep. Anim Nutr 2024;17:376-86. https://doi.org/10.1016/j.aninu.2024.03.003
  10. Zhang J, Meng S, Wang H, et al. Comparison of growth performance, carcass properties, fatty acid profile, and genes involved in fat metabolism in Nanyang and Landrace pigs. Genes 2024;15:186. https://doi.org/10.3390/genes15020186
  11. Schiavon S, Toscano A, Giannuzzi D, et al. Effects of slaughter weight and backfat depth on trimming, curing, and deboning losses and quality traits of Italian dry-cured ham. Food Res Int 2024;188:114450. https://doi.org/10.1016/j.foodres.2024.114450
  12. Qi W, Zhu S, Feng L, et al. Integrated analysis of the transcriptome and microbial diversity in the intestine of miniature pig obesity model. Microorganisms 2024;12:369. https://doi.org/10.3390/microorganisms12020369
  13. Choi YR, Kim YS, Kim MJ. Cinnamyl alcohol attenuates adipogenesis in 3T3-L1 cells by arresting the cell cycle. Int J Mol Sci 2024;25:693. https://doi.org/10.3390/ijms25020693
  14. Lin ZZ, Li ZQ, Li JJ, et al. Mfsd2a promotes the proliferation, migration, differentiation and adipogenesis of chicken intramuscular preadipocytes. Braz J Poult Sci 2022;24:eRBCA2021-1547. https://doi.org/10.1590/1806-9061-2021-1547
  15. Zhang XH, Li JX, Wu XX, et al. PABPN1 functions as a downstream gene of CREB to inhibit the proliferation of preadipocytes. Anim Biosci 2025;38:41-53. https://doi.org/10.5713/ab.24.0072
  16. Jia Z, Jin Z, Li M, et al. E2F transcription factor 5, a new regulator in adipogenesis to mediate the role of Krüppel-like factor 7 in chicken preadipocyte differentiation and proliferation. Poult Sci 2024;103:103728. https://doi.org/10.1016/j.psj.2024.103728
  17. Yang M, Jiang J, Ren R, Gao N, He J, Zhang Y. Role of ADAR1 on proliferation and differentiation in porcine preadipocytes. Animals 2024;14:1201. https://doi.org/10.3390/ani14081201
  18. Pan AL, Audrain M, Sakakibara E, et al. Dual-specificity protein phosphatase 6 (DUSP6) overexpression reduces amyloid load and improves memory deficits in male 5xFAD mice. Front Aging Neurosci 2024;16:1400447. https://doi.org/10.3389/fnagi.2024.1400447
  19. Png CW, Weerasooriya M, Li H, et al. DUSP6 regulates Notch1 signalling in colorectal cancer. Nat Commun 2024; 15:10087. https://doi.org/10.1038/s41467-024-54383-y
  20. Jiang C, Saiki Y, Hirota S, et al. Ablation of dual-specificity phosphatase 6 protects against nonalcoholic fatty liver disease via cytochrome P450 4A and mitogen-activated protein kinase. Am J Pathol 2023;193:1988-2000. https://doi.org/10.1016/j.ajpath.2023.09.003
  21. Ma Y, Liu Y, He S, Jiang Z. DUSP6: potential interactions with FXR1P in the nervous system. Indian J Biochem Biophys 2022;59:229-37.
  22. Ahmad MK, Abdollah NA, Shafie NH, Yusof NM, Razak SRA. Dual-specificity phosphatase 6 (DUSP6): a review of its molecular characteristics and clinical relevance in cancer. Cancer Biol Med 2018;15:14-28. https://doi.org/10.20892/j.issn.2095-3941.2017.0107
  23. Lin MC, Peng ZY, Chou HC, et al. Fecal protein analysis of Dusp6 knockout C57BL/6J mice by metaproteomics. Appl Biochem Biotechnol 2023;195:4215-36. https://doi.org/10.1007/s12010-023-04336-w
  24. Liu R, Peters M, Urban N, Knowlton J, Napierala T, Gabrysiak J. Mice lacking DUSP6/8 have enhanced ERK1/2 activity and resistance to diet-induced obesity. Biochem Biophys Res Commun 2020;533:17-22. https://doi.org/10.1016/j.bbrc.2020.08.106
  25. Xu H, Yang Q, Shen M, et al. Dual specificity MAPK phosphatase 3 activates PEPCK gene transcription and increases gluconeogenesis in rat hepatoma cells. J Biol Chem 2005;280: 36013-8. https://doi.org/10.1074/jbc.M508027200
  26. Feng B, Jiao P, Helou Y, et al. Mitogen-activated protein kinase phosphatase 3 (MKP-3)–deficient mice are resistant to dietinduced obesity. Diabetes 2014;63:2924-34. https://doi.org/10.2337/db14-0066
  27. Laragione T, Harris C, Rice N, Gulko PS. DUSP6 deletion protects mice and reduces disease severity in autoimmune arthritis. iScience 2024;27:110158. https://doi.org/10.1016/j.isci.2024.110158
  28. Jiang H, Deng L, Lin Z, et al. GSDMB interacts with IGF2BP1 to suppress colorectal cancer progression by modulating DUSP6-ERK pathway. Int Immunopharmacol 2024;143: 113280. https://doi.org/10.1016/j.intimp.2024.113280
  29. Rodrigues BA, Muñoz VR, Kuga GK, et al. Obesity increases mitogen-activated protein kinase phosphatase-3 levels in the hypothalamus of mice. Front Cell Neurosci 2017;11:313. https://doi.org/10.3389/fncel.2017.00313
  30. Feng B, He Q, Xu H. FOXO1-dependent up-regulation of MAP kinase phosphatase 3 (MKP-3) mediates glucocorticoid-induced hepatic lipid accumulation in mice. Mol Cell Endocrinol 2014;393:46-55. https://doi.org/10.1016/j.mce.2014.06.001
  31. Zhao Y, Pan J, Cao C, et al. RNF20 affects porcine adipocyte differentiation via regulation of mitotic clonal expansion. Cell Prolif 2021;54:e13131. https://doi.org/10.1111/cpr.13131
  32. Lefterova MI, Lazar MA. New developments in adipogenesis. Trends Endocrinol Metab 2009;20:107-14. https://doi.org/10.1016/j.tem.2008.11.005
  33. Moseti D, Regassa A, Kim WK. Molecular regulation of adipogenesis and potential anti-adipogenic bioactive molecules. Int J Mol Sci 2016;17:124. https://doi.org/10.3390/ijms17010124
  34. Brun RP, Kim JB, Hu E, Altiok S, Spiegelman BM. Adipocyte differentiation: a transcriptional regulatory cascade. Curr Opin Cell Biol 1996;8:826-32. https://doi.org/10.1016/S0955-0674(96)80084-6
  35. Tontonoz P, Spiegelman BM. Fat and beyond: the diverse biology of PPARγ. Annu Rev Biochem 2008;77:289-312. https://doi.org/10.1146/annurev.biochem.77.061307.091829
  36. Wojciechowicz T, Kolodziejski PA, Billert M, Strowski MZ, Nowak KW, Skrzypski M. The effects of neuropeptide B on proliferation and differentiation of porcine white preadipocytes into mature adipocytes. Int J Mol Sci 2023;24:6096. https://doi.org/10.3390/ijms24076096
  37. Mersmann HJ, Ding ST. Fatty acids modulate porcine adipocyte differentiation and transcripts for transcription factors and adipocyte-characteristic proteins. J Nutr Biochem 2001;12:101-8. https://doi.org/10.1016/S0955-2863(00)00136-4
  38. Pfuhlmann K, Pfluger PT, Schriever SC, Müller TD, Tschöp MH, Stemmer K. Dual specificity phosphatase 6 deficiency is associated with impaired systemic glucose tolerance and reversible weight retardation in mice. PLOS ONE 2017;12: e0183488. https://doi.org/10.1371/journal.pone.0183488
  39. Liu X, Qin K, Wang C, et al. Butyric acid reduced lipid deposition in immortalized chicken preadipocyte by inhibiting cell proliferation and differentiation. Poult Sci 2024;103: 104171. https://doi.org/10.1016/j.psj.2024.104171
  40. Bravo R, Frank R, Blundell PA, Macdonald-Bravo H. Cyclin/PCNA is the auxiliary protein of DNA polymerase-δ. Nature 1987;326:515-7. https://doi.org/10.1038/326515a0
  41. Juríková M, Danihel Ľ, Polák Š, Varga I. Ki67, PCNA, and MCM proteins: markers of proliferation in the diagnosis of breast cancer. Acta Histochem 2016;118:544-52. https://doi.org/10.1016/j.acthis.2016.05.002
  42. Krek W, Nigg EA. Differential phosphorylation of vertebrate p34cdc2 kinase at the G1/S and G2/M transitions of the cell cycle: identification of major phosphorylation sites. EMBO J 1991;10:305-16. https://doi.org/10.1002/j.1460-2075.1991.tb07951.x
  43. Su TT, Stumpff J. Promiscuity rules? The dispensability of cyclin E and Cdk2. Sci Stke 2004;2004:pe11. https://doi.org/10.1126/stke.2242004pe11
  44. Gu Y, Li D, Luo Q, et al. MicroRNA-145 inhibits human papillary cancer TPC1 cell proliferation by targeting DUSP6. Int J Clin Exp Med 2015;8:8590-8.
  45. Wu QN, Liao YF, Lu YX, et al. Pharmacological inhibition of DUSP6 suppresses gastric cancer growth and metastasis and overcomes cisplatin resistance. Cancer Lett 2018;412:243-55. https://doi.org/10.1016/j.canlet.2017.10.007
  46. Song H, Wu C, Wei C, et al. Silencing of DUSP6 gene by RNAi-mediation inhibits proliferation and growth in MDAMB-231 breast cancer cells: an in vitro study. Int J Clin Exp Med 2015;8:10481-90.
  47. Kim WK, Oh KJ, Choi HR, et al. MAP kinase phosphatase 3 inhibits brown adipocyte differentiation via regulation of Erk phosphorylation. Mol Cell Endocrinol 2015;416:70-6. https://doi.org/10.1016/j.mce.2015.08.023