苏淮猪背最长肌FAPs细胞体外成脂能力及其基因表达模式的研究

doi:10.11843/j.issn.0366-6964.2023.10.012

Abstract

Abstract: This study aimed to obtain higher purity of fibro/adipogenic progenitors (FAPs) from longissimus dorsi muscle of Suhuai pig, and evaluate the alteration of the adipogenic capacity and gene expression patterns during in vitro culture. The longissimus dorsi muscle of one-day Suhuai boar was digested to obtain suspended mixed cells. In order to determine the optimal differential adhesion time for FAPs cell, the purity of cell isolated from 4 different adhesion time were evaluated by immunofluorescence using an antibody against platelet-derived growth factor receptor α (PDGFRα). Then, the adipogenic differentiation ability of different passage of FAPs cells (P1, P3 and P5) cultured in vitro was compared, and the RNA-Seq was used to identify the differentially expressed genes (DEGs), KEGG pathways and GO functional classification in FAPs cells with different passages.The results showed that the purity of FAPs cells isolated from longissimus dorsi muscle of Suhuai pig by differential adherence for 30 min was much higher than other conditions. Evaluation of the adipogenic capacity of FAPs cells with different passages showed that the adipogenic ability of FAPs cells decreased significantly with the increase of passages in vitro. RNA-Seq results showed that the gene expression patterns of P3 and P5 generation FAPs cells were similar. Compared with P1 to P3 or P1 to P5 generation cells respectively, a total of 2 336 common DEGs was identified, including 1 102 up-regulated DEGs and 1 234 down-regulated DEGs. KEGG and GO analysis showed that the common up-regulated DEGs were mainly enriched in PI3K-AKT-FoxO, PI3K-AKT-HIF-1 and cGMP-PKG signaling pathways etc., and the common down-regulated DEGs were mainly enriched in DNA replication and repair signaling pathways etc. Among them, the genes of IRS1, FOXO3, CCNG2, EGFR and FGF1, which could inhibit cell proliferation and adipogenic differentiation were up-regulated, while the genes of CSF1R and SIPA1L2, which could promote cell proliferation were down-regulated. Compared P3 with P5 generation cells, the up-regulated DEGs were mainly enriched in NOD-like, MAPK and non-canonical Wnt signaling pathways etc.; The down-regulated DEGs were mainly enriched in VEGF, PPAR, Notch and IL-17 signaling pathways etc. Among them, TNFAIP3, AREG, FGF1 and FGF9 genes, which could inhibit adipogenic differentiation were up-regulated, and PPARγ, C/EBPα and LCN2 genes, which could promote adipogenic differentiation were down-regulated.These results showed that porcine FAPs cells with higher purity can be obtained by differential adherence for 30 min. The adipogenic capacity of porcine FAPs is continuously decreased during in vitro culture, and the gene expression patterns also undergo significant changes during in vitro culture. The major change for in vitro cultured FAPs is the reduction of gene expression related to cell proliferation and adipogenic differentiation. This study could provide a reference for developing a medium formula, which could maintain the adipogenic capacity of porcine FAPs cells during in vitro culture.

Key words: pig, FAPs cell, in vitro, adipogenic capacity, gene expression

CLC Number:

S828.2

JI Zhengyu, NI Mengru, ZHANG Zhaobo, ZHAO Gan, HUANG Zan, LI Pinghua, HUANG Ruihua, HOU Liming. Research on Adipogenic Capacity and Gene Expression Pattern of in Vitro FAPs Cells Derived from Longissimus Dorsi Muscle of Suhuai Pig[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4126-4142.

References 41

[1]	WOOD J D, ENSER M, FISHER A V, et al.Fat deposition, fatty acid composition and meat quality:a review[J].Meat Sci, 2008, 78(4):343-358.
[2]	DU M, WANG B, FU X, et al.Fetal programming in meat production[J].Meat Sci, 2015, 109:40-47.
[3]	JOE A W B, YI L, NATARAJAN A, et al.Muscle injury activates resident fibro/adipogenic progenitors that facilitate myogenesis[J].Nat Cell Biol, 2010, 12(2):153-163.
[4]	UEZUMI A, FUKADA S, YAMAMOTO N, et al.Identification and characterization of PDGFRα+ mesenchymal progenitors in human skeletal muscle[J].Cell Death Dis, 2014, 5(4):e1186.
[5]	DU M, HUANG Y, DAS A K, et al.Meat Science and Muscle Biology Symposium:manipulating mesenchymal progenitor cell differentiation to optimize performance and carcass value of beef cattle[J].J Anim Sci, 2013, 91(3):1419-1427.
[6]	LI X, FU X, YANG G, et al.Review:enhancing intramuscular fat development via targeting fibro-adipogenic progenitor cells in meat animals[J].Animal, 2020, 14(2):312-321.
[7]	AGLEY C C, ROWLERSON A M, VELLOSO C P, et al.Human skeletal muscle fibroblasts, but not myogenic cells, readily undergo adipogenic differentiation[J].J Cell Sci, 2013, 126(Pt 24):5610-5625.
[8]	KRUK Z A, BOTTEMA M J, REYES-VELIZ L, et al.Vitamin A and marbling attributes:intramuscular fat hyperplasia effects in cattle[J].Meat Sci, 2018, 137:139-146.
[9]	CHEN F F, WANG Y Q, TANG G R, et al.Differences between porcine longissimus thoracis and semitendinosus intramuscular fat content and the regulation of their preadipocytes during adipogenic differentiation[J].Meat Sci, 2019, 147:116-126.
[10]	MOLINA T, FABRE P, DUMONT N A.Fibro-adipogenic progenitors in skeletal muscle homeostasis, regeneration and diseases[J].Open Biol, 2021, 11(12):210110.
[11]	DENG B, ZHANG F, WEN J H, et al.The transcriptomes from two adipocyte progenitor cell types provide insight into the differential functions of MSTN[J].Genomics, 2020, 112(5):3826-3836.
[12]	AYKAN A, OZTURK S, SAHIN I, et al.The effects of hydrogen sulfide on adipocyte viability in human adipocyte and adipocyte-derived mesenchymal stem cell cultures under ischemic conditions[J].Ann Plast Surg, 2015, 75(6):657-665.
[13]	CHAPMAN M A, MUKUND K, SUBRAMANIAM S, et al.Three distinct cell populations express extracellular matrix proteins and increase in number during skeletal muscle fibrosis[J].Am J Physiol Cell Physiol, 2017, 312(2):C131-C143.
[14]	JUDSON R N, LOW M, EISNER C, et al.Isolation, culture, and differentiation of fibro/adipogenic progenitors (FAPs) from skeletal muscle[M]//RYALL J G.Skeletal Muscle Development.New York:Humana Press, 2017:93-103.
[15]	UEZUMI A, FUKADA S I, YAMAMOTO N, et al.Mesenchymal progenitors distinct from satellite cells contribute to ectopic fat cell formation in skeletal muscle[J].Nat Cell Biol, 2010, 12(2):143-152.
[16]	WOSCZYNA M N, BISWAS A A, COGSWELL C A, et al.Multipotent progenitors resident in the skeletal muscle interstitium exhibit robust BMP-dependent osteogenic activity and mediate heterotopic ossification[J].J Bone Miner Res, 2012, 27(5):1004-1017.
[17]	ALECOVA B, GATTO S, ETXANIZ U, et al.Dynamics of cellular states of fibro-adipogenic progenitors during myogenesis and muscular dystrophy[J].Nat Commun, 2018, 9(1):3670.
[18]	TAHA M F, HEDAYATI V.Isolation, identification and multipotential differentiation of mouse adipose tissue-derived stem cells[J].Tissue Cell, 2010, 42(4):211-216.
[19]	CHEN M, CHOI S, WEN T, et al.A p53-phosphoinositide signalosome regulates nuclear AKT activation[J].Nat Cell Biol, 2022, 24(7):1099-1113.
[20]	ZHANG H B, DOMMA A J, GOODRUM F D, et al.The akt forkhead box O transcription factor axis regulates human cytomegalovirus replication[J].mBio, 2022, 13(4):e0104222.
[21]	BLOEDJES T A, DE WILDE G, MAAS C, et al.AKT signaling restrains tumor suppressive functions of FOXO transcription factors and GSK3 kinase in multiple myeloma[J].Blood Adv, 2020, 4(17):4151-4164.
[22]	GARRIDO-SÁNCHEZ L, DEL MAR ROCA-RODRÍGUEZ M, FERNÁNDEZ-VELEDO S, et al.CCNG2 and CDK4 is associated with insulin resistance in adipose tissue[J].Surg Obes Relat Dis, 2014, 10(4):691-696.
[23]	ZHONG H, CHILES K, FELDSER D, et al.Modulation of hypoxia-inducible factor 1alpha expression by the epidermal growth factor/phosphatidylinositol 3-kinase/PTEN/AKT/FRAP pathway in human prostate cancer cells:implications for tumor angiogenesis and therapeutics[J].Cancer Res, 2000, 60(6):1541-1545.
[24]	REGAZZETTI C, PERALDI P, GRÉMEAUX T, et al.Hypoxia decreases insulin signaling pathways in adipocytes[J]. Diabetes, 2009, 58(1):95-103.
[25]	WANG T, WANG Y X, YAMASHITA H.Evodiamine inhibits adipogenesis via the EGFR-PKCα-ERK signaling pathway[J]. FEBS Lett, 2009, 583(22):3655-3659.
[26]	WANG S P, CAO S, ARHATTE M, et al.Adipocyte Piezo1 mediates obesogenic adipogenesis through the FGF1/FGFR1 signaling pathway in mice[J].Nat Commun, 2020, 11(1):2303.
[27]	JONKER J W, SUH J M, ATKINS A R, et al.A PPARγ-FGF1 axis is required for adaptive adipose remodelling and metabolic homeostasis[J].Nature, 2012, 485(7398):391-394.
[28]	SCHALL N, GARCIA J J, KALYANARAMAN H, et al.Protein kinase G1 regulates bone regeneration and rescues diabetic fracture healing[J].JCI Insight, 2020, 5(9):e135355.
[29]	O'BRIEN J, MARTINSON H, DURAND-ROUGELY C, et al.Macrophages are crucial for epithelial cell death and adipocyte repopulation during mammary gland involution[J].Development, 2012, 139(2):269-275.
[30]	CHEN T Q, PENG Y, HU W J, et al.Irisin enhances chondrogenic differentiation of human mesenchymal stem cells via Rp1/PI3K/AKT axis[J].Stem Cell Res Ther, 2022, 13(1):392.
[31]	GENG X, IRVIN M R, HIDALGO B, et al.An exome-wide sequencing study of lipid response to high-fat meal and fenofibrate in Caucasians from the GOLDN cohort[J].J Lipid Res, 2018, 59(4):722-729.
[32]	AI L Y, XU Q Q, WU C W, et al.A20 Attenuates FFAs-induced lipid accumulation in nonalcoholic steatohepatitis[J].Int J Biol Sci, 2015, 11(12):1436-1446.
[33]	AI L Y, WANG X H, CHEN Z W, et al.A20 reduces lipid storage and inflammation in hypertrophic adipocytes via p38 and Akt signaling[J].Mol Cell Biochem, 2016, 420(1-2):73-83.
[34]	ZACHARA M, RAINER P Y, HASHIMI H, et al.Mammalian adipogenesis regulator (Areg) cells use retinoic acid signalling to be non-and anti-adipogenic in age-dependent manner[J].EMBO J, 2022, 41(18):e108206.
[35]	BEENKEN A, MOHAMMADI M.The FGF family:biology, pathophysiology and therapy[J].Nat Rev Drug Discov, 2009, 8(3):235-253.
[36]	HUANG K, WANG Y, ZHU J J, et al.Regulation of fibroblast growth factor 9 on the differentiation of goat intramuscular adipocytes[J].Anim Sci J, 2021, 92(1):e13627.
[37]	MANCA C, PINTUS S, MURRU E, et al.Fatty acid metabolism and derived-mediators distinctive of PPAR-α activation in obese subjects post bariatric surgery[J].Nutrients, 2021, 13(12):4340.
[38]	TAKAHASHI J, TAKAHASHI N, TADAISHI M, et al.Valerenic acid promotes adipocyte differentiation, adiponectin production, and glucose uptake via its PPARγ ligand activity[J].ACS Omega, 2022, 7(51):48113-48120.
[39]	EL-JACK A K, HAMM J K, PILCH P F, et al. Reconstitution of insulin-sensitive glucose transport in fibroblasts requires expression of both PPARγ and C/EBPα[J].J Biol Chem, 1999, 274(12):7946-7951.
[40]	SHARMA N, KAUR R, YADAV B, et al.Transient delivery of A-C/EBP protein perturbs differentiation of 3T3-L1 cells and induces preadipocyte marker genes[J].Front Mol Biosci, 2020, 7:603168.
[41]	GUO H, BAZUINE M, JIN D Z, et al.Evidence for the regulatory role of lipocalin 2 in high-fat diet-induced adipose tissue remodeling in male mice[J].Endocrinology, 2013, 154(10):3525-3538.

Research on Adipogenic Capacity and Gene Expression Pattern of in Vitro FAPs Cells Derived from Longissimus Dorsi Muscle of Suhuai Pig

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