Please wait a minute...
Journal of Integrative Agriculture  2026, Vol. 25 Issue (9): 3786-3799    DOI: 10.1016/j.jia.2025.09.006
Animal Science · Veterinary Medicine Advanced Online Publication | Current Issue | Archive | Adv Search |
MFAP5 enhances the cold resistance of piglets by promoting the transition of adipocyte progenitor cells to fibroblast lineage

Xiangfei Ma1, Mengting Li1, Shengda Qiu1, Di Liu2, Hong Ma2, Wei Wei1, Lifan Zhang1, Zan Huang1#, Jie Chen1#

1 College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China

2 Institute of Animal Husbandry, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, China

 Highlights 

Dorsal subcutaneous adipose tissues (subWATs) in piglets exhibited thermogenic potential and fibrotic remodeling under cold stress.

Single-nucleus RNA-seq (snRNA-seq) and RNA-seq identified MFAP5 as a critical mediator of cold-induced adipose plasticity, which promoted fibrogenic lineage commitment in fibro/adipogenic progenitors (FAPs).

MFAP5 overexpression or conditioned medium inhibited adipocyte differentiation and induced fibroblast-like phenotypes in C3H10T1/2 and porcine stromal vascular fraction (SVF) cells, accompanied by elevated mitochondrial activity.

MFAP5 activates the Hippo signaling pathway, thereby increasing the nuclear translocation of YAP1 and upregulating fibrogenic markers such as ACTA2 and CTGF.

Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
摘要  

猪缺乏典型的棕色脂肪组织,但有研究表明猪脂肪细胞具有不依赖UCP1产热的能力,然而目前对这些产热脂肪细胞的发育过程和调控机制仍然缺乏了解。本研究探究了二花脸仔猪背部皮下脂肪组织在冷刺激下的适应性反应机制,重点关注纤维/脂肪生成祖细胞(FAPs)的分化命运转变及其调控分子微纤维相关蛋白5MFAP5)的作用,揭示了冷刺激条件下脂肪组织纤维化与产热功能的关联。本文以4对体重相近的6-8周龄雄性全同胞二花脸仔猪为试验材料,将全同胞二花脸仔猪随机分为室温组(28±0.5℃)和冷刺激组(11±0.5℃)进行30天试验。采集10个部位脂肪组织进行分析,发现背部皮下脂肪组织在冷激下表现出显著的产热潜力,同时观察到背部皮下脂肪组织发生组织结构重塑。综合分析背部皮下脂肪组织转录组测序和单细胞测序数据,发现冷刺激促进FAPs向非脂肪成纤维细胞分化,且MFAP5是冷刺激背部皮下脂肪组织可塑性的潜在调节因子。体外细胞试验证实,过表达MFAP5或添加MFAP5条件培养基抑制前脂肪细胞分化为成熟脂肪细胞,促进其向非脂肪成纤维细胞分化,并显著增强非脂肪成纤维化细胞的线粒体生物合成。进一步研究表明,MFAP5主要通过Hippo-YAP1信号通路发挥作用。综上所述,本研究发现冷刺激通过上调二花脸仔猪背部皮下脂肪组织中MFAP5表达,激活Hippo-YAP1信号通路,促使FAPs分化为非脂肪成纤维细胞,进而抑制脂肪生成并促进脂肪组织纤维化和产热适应。这一发现揭示了MFAP5作为冷适应调控因子的新功能,为理解哺乳动物脂肪组织可塑性提供了新视角。



Abstract  

Although pigs lack classical brown adipose tissue, several studies have demonstrated that porcine adipocytes possess the capacity to undergo thermogenesis through UCP1-independent mechanisms.  However, the developmental processes and regulatory mechanisms underlying these thermogenic adipocytes remain poorly characterized.  Here, we found that dorsal subcutaneous adipose tissues (subWATs) in pigs exhibits significant thermogenic potential under cold stress.  Notably, we observed substantial cold-induced structural remodeling in dorsal subWATs, characterized by increased fibrotic deposition.  An integrated analysis of snRNA-seq and RNA-seq data on dorsal subWATs identified MFAP5, which encodes a microfibril-associated glycoprotein in the extracellular matrix, as a potential regulator of the cold-induced plasticity of dorsal subWATs.  Both MFAP5 overexpression and MFAP5-conditioned medium (MFAP5-CM) not only inhibited preadipocyte differentiation into adipocytes but also promoted their commitment to non-adipogenic fibrogenic lineages.  Furthermore, MFAP5 treatments significantly enhanced the mitochondrial biogenesis of these fibrogenic cells. Mechanistic investigations showed that these phenotypic alterations are predominantly mediated through the Hippo signaling pathway.  In summary, our findings elucidate the pivotal role of MFAP5 in regulating adipocyte development following cold exposure, thus providing crucial insights into the molecular mechanisms underlying porcine adaptation to cold stress.

Keywords:  piglets       cold adaptation        cell fate        microfibril associated protein 5  
Received: 17 February 2025   Accepted: 01 August 2025 Online: 04 September 2025  
Fund: 

This work was supported by grants from the Joint Funds of the National Natural Science Foundation of China (U20A2052), the National Natural Science Foundation of China (32372853, 32170847), the Jiangsu Agriculture Science and Technology Innovation Fund, China (CX(23)3137). 

About author:  Xiangfei Ma, E-mail: 2021205002@stu.njau.edu.cn; #Correspondence Jie Chen, E-mail: jiechen@njau.edu.cn; Zan Huang, E-mail: huangzan@njau.edu.cn

Cite this article: 

Xiangfei Ma, Mengting Li, Shengda Qiu, Di Liu, Hong Ma, Wei Wei, Lifan Zhang, Zan Huang, Jie Chen. 2026. MFAP5 enhances the cold resistance of piglets by promoting the transition of adipocyte progenitor cells to fibroblast lineage. Journal of Integrative Agriculture, 25(9): 3786-3799.

Berg F, Gustafson U, Andersson L. 2006. The uncoupling protein 1 gene (UCP1) is disrupted in the pig lineage: a genetic explanation for poor thermoregulation in piglets. PLoS Genetics, 2, e129.

Bornstein M R, Neinast M D, Zeng X, Chu Q, Axsom J, Thorsheim C, Li K, Blair M C, Rabinowitz J D, Arany Z. 2023. Comprehensive quantification of metabolic flux during acute cold stress in mice. Cell Metabolism35, 2077-2092.e2076.

Burl R B, Ramseyer V D, Rondini E A, Pique-Regi R, Lee Y H, Granneman J G. 2018. Deconstructing adipogenesis induced by β3-adrenergic receptor activation with single-cell expression profiling. Cell Metabolism28, 300-309.e304.

Cheng Y M, Hong P C, Song M M, Zhu H N, Qin J, Zhang Z D, Chen H, Ma X Z, Tian M Y, Zhu W Y, Huang Z. 2023. An immortal porcine preadipocyte cell strain for efficient production of cell-cultured fat. Communications Biology6, 1202.

Côté J A, Ostinelli G, Gauthier M F, Lacasse A, Tchernof A. 2019. Focus on dedifferentiated adipocytes: Characteristics, mechanisms, and possible applications. Cell and Tissue Research378, 385–398.

Cristancho A G, Lazar M A. 2011. Forming functional fat: A growing understanding of adipocyte differentiation. Nature Reviews. Molecular Cell Biology12, 722–734.

Cui H X, LUO N, Guo L P, Liu L, Xing S Y, Zhao G P, Wen J. 2023. TIMP2 promotes intramuscular fat deposition by regulating the extracellular matrix in chicken. Journal of Integrative Agriculture22, 853–863.

Datta R, Podolsky M J, Atabai K. 2018. Fat fibrosis: friend or foe? JCI Insight3, 19.

Le Dividich J, Noblet J. 1981. Colostrum intake and thermoregulation in the neonatal pig in relation to environmental temperature. Biology of the Neonate40, 167–174.

Du K, Ramachandran A, Mcgill M R, Mansouri A, Asselah T, Farhood A, Woolbright B L, Ding W X, Jaeschke H. 2017. Induction of mitochondrial biogenesis protects against acetaminophen hepatotoxicity. y. Food and Chemical Toxicology108, 339–350.

Ghesmati Z, Rashid M, Fayezi S, Gieseler F, Alizadeh E, Darabi M. 2024. An update on the secretory functions of brown, white, and beige adipose tissue: Towards therapeutic applications. Reviews in Endocrine & Metabolic Disorders25, 279–308.

Han C, Leonardo T R, Romana-Souza B, Shi J, Keiser S, Yuan H, Altakriti M, Ranzer M J, Ferri-Borgogno S, Mok S C, Koh T J, Hong S J, Chen L, Dipietro L A. 2023. Microfibril-associated protein 5 and the regulation of skin scar formation. Scientific Reports13, 8728.

He T, Wang S, Li S, Shen H, Hou L, Liu Y, Wei Y, Xie F, Zhang Z, Zhao Z, Mo C, Guo H, Huang Q, Zhang R, Shen D, Li B. 2023. Suppression of preadipocyte determination by SOX4 limits white adipocyte hyperplasia in obesity. iScience26, 106289.

Hoerst K, Van Den Broek L, Sachse C, Klein O, Von Fritschen U, Gibbs S, Hedtrich S. 2019. Regenerative potential of adipocytes in hypertrophic scars is mediated by myofibroblast reprogramming. Journal of Molecular Medicine (Berlin, Germany), 97, 761–775.

Hou L, Shi J, Cao L, Xu G, Hu C, Wang C. 2017. Pig has no uncoupling protein 1. Biochemical and Biophysical Research Communications487, 795–800.

Hou L, Xie M, Cao L, Shi J, Xu G, Hu C, Wang C. 2018. Browning of pig white preadipocytes by co-overexpressing pig PGC-1α and mice UCP1. Cellular Physiology and Biochemistry48, 556–568.

Huang J, Wu S, Barrera J, Matthews K, Pan D. 2005. The Hippo signaling pathway coordinately regulates cell proliferation and apoptosis by inactivating Yorkie, the Drosophila homolog of YAP. Cell122, 421–434.

Hwang J H, Kim K M, Oh H T, Yoo G D, Jeong M G, Lee H, Park J, Jeong K, Kim Y K, Ko Y G, Hwang E S, Hong J H. 2022. TAZ links exercise to mitochondrial biogenesis via mitochondrial transcription factor A. Nature Communications13, 653.

Ikeda K, Maretich P, Kajimura S. 2018. The common and distinct features of brown and beige adipocytes. Trends in Endocrinology and Metabolism29, 191–200.

Jang E H, Lee J H, Kim S A. 2021. Acute valproate exposure induces mitochondrial Biogenesis and autophagy with FOXO3a modulation in SH-SY5Y cells. Cells10, 2522.

Kelley K W, Blecha F, Regnier J A. 1982. Cold exposure and absorption of colostral immunoglobulins by neonatal pigs. Journal of Animal Science55, 363–368.

Kotarsky C J, Johnson N R, Mahoney S J, Mitchell S L, Schimek R L, Stastny S N, Hackney K J. 2021. Time-restricted eating and concurrent exercise training reduces fat mass and increases lean mass in overweight and obese adults. Physiolgical Reports9, e14868.

Krois C R, Vuckovic M G, Huang P, Zaversnik C, Liu C S, Gibson C E, Wheeler M R, Obrochta K M, Min J H, Herber C B, Thompson A C, Shah I D, Gordon S P, Hellerstein M K, Napoli J L. 2019. RDH1 suppresses adiposity by promoting brown adipose adaptation to fasting and re-feeding. Cellular and Molecular Life Sciences76, 2425–2447.

Lemaire R, Bayle J, Mecham R P, Lafyatis R. 2007. Microfibril-associated MAGP-2 stimulates elastic fiber assembly. The Journal of Biological Chemistry282, 800–808.

Li P H, Ma X, Zhang Y Q, Zhang Q, Huang R H. 2017. Progress in the physiological and genetic mechanisms underlying the high prolificacy of the Erhualian pig. Hereditas39, 1016–1024. (in Chinese)

Lin D, Chun T H, Kang L. 2016. Adipose extracellular matrix remodelling in obesity and insulin resistance. Biochemical Pharmacology119, 8–16.

Lin J, Cao C, Tao C, Ye R, Dong M, Zheng Q, Wang C, Jiang X, Qin G, Yan C, Li K, Speakman J R, Wang Y, Jin W, Zhao J. 2017. Cold adaptation in pigs depends on UCP3 in beige adipocytes. Journal of Molecular Cell Biology9, 364–375.

Lin W, Tang Y, Zhao Y, Zhao J, Zhang L, Wei W, Chen J. 2020. MiR-144-3p targets foxo1 to reduce its regulation of adiponectin and promote adipogenesis. Frontiers in Genetics11, 603144.

Mammoto A, Muyleart M, Kadlec A, Gutterman D, Mammoto T. 2018. YAP1-TEAD1 signaling controls angiogenesis and mitochondrial biogenesis through PGC1α. Microvascular Research119, 73–83.

Maniyadath B, Zhang Q, Gupta R K, Mandrup S. 2023. Adipose tissue at single-cell resolution. Cell Metabolism35, 386–413.

Miller C N, Yang J Y, England E, Yin A, Baile C A, Rayalam S. 2015. Isoproterenol increases uncoupling, glycolysis, and markers of beiging in mature 3T3-L1 adipocytes. PLoS ONE10, e0138344.

Newman A A C, Serbulea V, Baylis R A, Shankman L S, Bradley X, Alencar G F, Owsiany K, Deaton R A, Karnewar S, Shamsuzzaman S, Salamon A, Reddy M S, Guo L, Finn A, Virmani R, Cherepanova O A, Owens G K. 2021. Multiple cell types contribute to the atherosclerotic lesion fibrous cap by PDGFRβ and bioenergetic mechanisms. Nature Metabolism3, 166–181.

Norreen-Thorsen M, Struck E C, Öling S, Zwahlen M, Von Feilitzen K, Odeberg J, Lindskog C, Pontén F, Uhlén M, Dusart P J, Butler L M. 2022. A human adipose tissue cell-type transcriptome atlas. Cell Reports40, 111046.

O’mara A E, Johnson J W, Linderman J D, Brychta R J, Mcgehee S, Fletcher L A, Fink Y A, Kapuria D, Cassimatis T M, Kelsey N, Cero C, Sater Z A, Piccinini F, Baskin A S, Leitner B P, Cai H, Millo C M, Dieckmann W, Walter M, Javitt N B, et al. 2020. Chronic mirabegron treatment increases human brown fat, HDL cholesterol, and insulin sensitivity. The Jounal of Clinical Investigation130, 2209–2219.

Peng Z, Ren Z, Tong Z, Zhu Y, Zhu Y, Hu K. 2023. Interactions between MFAP5 + fibroblasts and tumor-infiltrating myeloid cells shape the malignant microenvironment of colorectal cancer. Journal of Translational Medicine21, 405.

Rockey D C, Weymouth N, Shi Z. 2013. Smooth muscle α actin (Acta2) and myofibroblast function during hepatic wound healing. PLoS ONE8, e77166.

Shao M, Wang Q A, Song A, Vishvanath L, Busbuso N C, Scherer P E, Gupta R K. 2019. Cellular origins of beige fat cells revisited. Diabetes68, 1874–1885.

Shook B A, Wasko R R, Mano O, Rutenberg-Schoenberg M, Rudolph M C, Zirak B, Rivera-Gonzalez G C, López-Giráldez F, Zarini S, Rezza A, Clark D A, Rendl M, Rosenblum M D, Gerstein M B, Horsley V. 2020. Dermal adipocyte lipolysis and myofibroblast conversion are required for efficient skin repair. Cell Stem Cell26, 880-895.e886.

Tamura T, Kodama T, Sato K, Murai K, Yoshioka T, Shigekawa M, Yamada R, Hikita H, Sakamori R, Akita H, Eguchi H, Johnson R L, Yokoi H, Mukoyama M, Tatsumi T, Takehara T. 2021. Dysregulation of PI3K and Hippo signaling pathways synergistically induces chronic pancreatitis via CTGF upregulation. The Journal of Clinical Investigation131, 13.

Trayhurn P, Temple N J, Van Aerde J. 1989. Evidence from immunoblotting studies on uncoupling protein that brown adipose tissue is not present in the domestic pig. Canadian Journal of Physiology and Pharmacology67, 1480–1485.

Vaittinen M, Kolehmainen M, Rydén M, Eskelinen M, Wabitsch M, Pihlajamäki J, Uusitupa M, Pulkkinen L. 2015. MFAP5 is related to obesity-associated adipose tissue and extracellular matrix remodeling and inflammation. Obesity (Silver Spring), 23, 1371–1378.

Valenzi E, Bulik M, Tabib T, Morse C, Sembrat J, Trejo Bittar H, Rojas M, Lafyatis R. 2019. Single-cell analysis reveals fibroblast heterogeneity and myofibroblasts in systemic sclerosis-associated interstitial lung disease. Annals of the Rheumatic Diseases78, 1379–1387.

Vijay J, Gauthier M F, Biswell R L, Louiselle D A, Johnston J J, Cheung W A, Belden B, Pramatarova A, Biertho L, Gibson M, Simon M M, Djambazian H, Staffa A, Bourque G, Laitinen A, Nystedt J, Vohl M C, Fraser J D, Pastinen T, Tchernof A, et al. 2020. Single-cell analysis of human adipose tissue identifies depot and disease specific cell types. Nature Metabolism2, 97–109.

Villarroya F, Cereijo R, Villarroya J, Giralt M. 2017. Brown adipose tissue as a secretory organ. Nature Reviews: Endocrinology13, 26–35.

Winther S, Isidor M S, Basse A L, Skjoldborg N, Cheung A, Quistorff B, Hansen J B. 2018. Restricting glycolysis impairs brown adipocyte glucose and oxygen consumption. American Journal of Physiology. Endocrinology and Metabolism314, E214–E223.

Wu Y, Li C S, Meng R Y, Jin H, Chai O H, Kim S M. 2024. Regulation of Hippo-YAP/CTGF signaling by combining an HDAC inhibitor and 5-fluorouracil in gastric cancer cells. Toxicology and Applied Pharmacology482, 116786.

Xiao F, Jiang H, Li Z, Jiang X, Chen S, Niu Y, Yin H, Shu Y, Peng B, Lu W, Li X, Li Z, Lan S, Xu X, Guo F. 2023. Reduced hepatic bradykinin degradation accounts for cold-induced BAT thermogenesis and WAT browning in male mice. Nature Communications, 14, 2523.

Xie X, Huang C, Huang Y, Zou X, Zhou R, Ai H, Huang L, Ma J. 2023. Genetic architecture for skeletal muscle glycolytic potential in Chinese Erhualian pigs revealed by a genome-wide association study using 1.4M SNP array. Frontiers in Genetics14, 1141411.

Yeung T L, Leung C S, Yip K P, Sheng J, Vien L, Bover L C, Birrer M J, Wong S T C, Mok S C. 2019. Anticancer immunotherapy by MFAP5 blockade inhibits fibrosis and enhances chemosensitivity in ovarian and pancreatic cancer. Clinical Cancer Research25, 6417–6428.

Zeve D, Seo J, Suh J M, Stenesen D, Tang W, Berglund E D, Wan Y, Williams L J, Lim A, Martinez M J, Mckay R M, Millay D P, Olson E N, Graff J M. 2012. Wnt signaling activation in adipose progenitors promotes insulin-independent muscle glucose uptake. Cell Metabolism15, 492–504.

Zhang C, Fu Q, Shao K, Liu L, Ma X, Zhang F, Zhang X, Meng L, Yan C, Zhao X. 2022. Indole-3-acetic acid improves the hepatic mitochondrial respiration defects by PGC1a up-regulation. Cellular Signalling99, 110442.

Zhang T, Li H, Sun S, Zhou W, Zhang T, Yu Y, Wang Q, Wang M. 2023. Microfibrillar-associated protein 5 suppresses adipogenesis by inhibiting essential coactivator of PPARγ. Scientific Reports13, 5589.

Zheng Q, Lin J, Huang J, Zhang H, Zhang R, Zhang X, Cao C, Hambly C, Qin G, Yao J, Song R, Jia Q, Wang X, Li Y, Zhang N, Piao Z, Ye R, Speakman J R, Wang H, Zhou Q, et al. 2017. Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity. Proceedings of the National Academy of Sciences of the United States of America114, e9474–e9482.

[1] Yetong Xu, Chengyu Zhou, Yingying Lu, Xutong Guo, Minyue Zong, Junwei Zhu, Pan Zhou, Jiaman Pang, Xie Peng, Zhihong Sun. Multi-omic analysis for dietary supplementation of different ratios of soluble and insoluble fiber on intestinal microbiota, metabolites and inflammation of weaned piglets[J]. >Journal of Integrative Agriculture, 2026, 25(4): 1606-1618.
[2] WANG Man, YU Bing, HE Jun, YU Jie, LUO Yu-heng, LUO Jun-qiu, MAO Xiang-bin, CHEN Dai-wen. The toxicological effect of dietary excess of saccharicterpenin, the extract of camellia seed meal, in piglets[J]. >Journal of Integrative Agriculture, 2020, 19(1): 211-224.
[3] ZHU Cui, WANG Li, WEI Shao-yong, CHEN Zhuang, MA Xian-yong, ZHENG Chun-tian, JIANG Zongyong. Effect of yeast Saccharomyces cerevisiae supplementation on serum antioxidant capacity, mucosal sIgA secretions and gut microbial populations in weaned piglets[J]. >Journal of Integrative Agriculture, 2017, 16(09): 2029-2037.
[4] ZHU Cui, GUO Chang-yi, GAO Kai-guo, WANG Li, CHEN Zhuang, MA Xian-yong, JIANG Zong-yong . Dietary arginine supplementation in multiparous sows during lactation improves the weight gain of suckling piglets[J]. >Journal of Integrative Agriculture, 2017, 16(03): 648-655.
[5] ZHANG Tian, WANG Li-gang, SHI Hui-bi, YAN Hua, ZHANG Long-chao, LIU Xin, PU Lei, LIANG Jing, ZHANG Yue-bo, ZHAO Ke-bin, WANG Li-xian . Heritabilities and genetic and phenotypic correlations of litter uniformity and litter size in Large White sows[J]. >Journal of Integrative Agriculture, 2016, 15(4): 848-854.
[6] HANG Su-qin , ZHU Wei-yun. Gut Bacterial and Lactobacilli Communities of Weaning Piglets in Response to Mannan Oligosaccharide and Sugar Beet Pulp In vitro Fermentation[J]. >Journal of Integrative Agriculture, 2012, 12(1): 122-133.
[7] WANG Xiu-qi, FENG You, SHU Gang, JIANG Qing-yan, YANG Jing-pei, ZHANG Zi-feng. Effect of Dietary Supplementation with Hydrolyzed Wheat Gluten on Growth Performance, Cell Immunity and Serum Biochemical Indices of Weaned Piglets (Sus scrofa)[J]. >Journal of Integrative Agriculture, 2011, 10(6): 938-945.
No Suggested Reading articles found!