Abstract:In order to investigate the differential gene expression on intramuscular and subcutaneous preadipocytes during differentiation in Congjiang Xiang pig (Sus scrofa), the fat precursors culture model in vitro of Congjiang Xiang pig was established. In this study, Congjiang Xiang pig in five days old was provided, then subcutaneous adipose tissue and the longissimus dorsi muscle was excised. We used collagenase to harvest preadipocytes from longissimus dorsi muscle and subcutaneous adipose tissue respectively. The two preadipocytes were induced differentiation by a series of hormone-induced complexes. The mRNA expression levels of gene involved in adipogenesis, such as peroxisome proliferator-activated receptorsγ (PPARγ), lipoprotein lipase (LPL), fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), adipose triglyceride lipase (ATGL) and Hormone sensitive lipase (HSL), were compared by real-time PCR in the two preadipocytes. The result showed that the model of preadipocytes in vitro was successfully established from Congjiang pig. The mRNA expression levels of PPARγ, FAS and ACC in intramuscular preadipocytes were significantly a higher at 72 h after the addition of inducer, which a higher than the level in 0 h (P<0.05). The mRNA expression level of LPL was no significant difference in the various stages, but there was a significant up-regulation of LPL in 144 h after the addition of inducer, and the value was significantly higher than the other stages (P<0.01). The mRNA expression levels of ATGL and HSL showed the pattern of “up-down-up”in the differentiation phase. In the subcutaneous preadipocytes, the mRNA expression level of PPARγ was the highest in 72 h after induction, and the value was significantly higher than in other stages (P<0.01). The mRNA expression levels of LPL, FAS, ACC, ATGL and HSL were all higher in 48 and 72 h, and the value was significantly higher than that in 0h (P<0.01). In summary, this study showed that in intramuscular preadipocytes, the genes related to adipocyte differentiation were more active at the late stage of differentiation, while in subcutaneous preadipocytes, the related genes were more active at the earlier stage of differentiation in vitro. The result prompt that the formation of subcutaneous adipose is much earlier than intramuscular adipose, which may be of great significance in fat deposition mechanism of Congjiang Xiang pig for further study.
[1] Rosen E D. The transcriptional basis of adipocyte development.[J]. Prostaglandins Leukotrienes & Essential Fatty Acids, 2005, 73(1):31-4.[2] Zhang F, Deng B, Wen J, et al. PPARγ and MyoD are differentially regulated by myostatin in adipose-derived stem cells and muscle satellite cells.[J]. Biochemical & Biophysical Research Communications, 2015, 458(2):375-380.[3] Cliona M. Stapleton, Douglas G. Mashek, Shuli Wang,etal. Lysophosphatidic Acid Activates Peroxisome Proliferator Activated Receptor-γ in CHO Cells That Over-Express Glycerol 3-Phosphate Acyltransferase-1[J]. Plos One, 2011, 6(4):e18932-e18932.[4] Shen W J, Yu Z, Patel S, et al. Hormone-Sensitive Lipase Modulates Adipose Metabolism Through PPARγ[J]. Biochimica Et Biophysica Acta, 2011, 1811(1):9-16.[5] Ramirez-Zacarias JL, Castro-Mufiozledo F and kuri-Harcuch W. Quantitation of adipose conversion and triglycerides by staining intracyctoplasmic lipids with Oil red 0[J]. Histochemistry,1992, 97:493-497.[6]马晶晶,章涛. PPARγ功能与疾病关系研究进展[J]. 中国药理学通报,2012,05:601-604.[7] Noh JR, Kim YH, Hwang JH,et al. Scoparone inhibits adipocyte differentiation through down-regulation of peroxisome proliferators-activated receptor γ in 3T3-L1 preadipocytes.[J]. Food Chemistry, 2013, 141(2):723-730.[8] Linhart H G, Ishimura-Oka K, Demayo F, et al. C/EBPalpha is required for differentiation of white, but not brown, adipose tissue.[J]. Proceedings of the National Academy of Sciences, 2001, 98(22):12532-7.[9] Johnson A M F, Olefsky J M. The origins and drivers of insulin resistance.[J]. Cell, 2013, 152(4):673-84.[10] Tontonoz P, Spiegelman B M. Fat and Beyond: The Diverse Biology of PPARγ[J]. Biochemistry, 2008, 77(77):289-312.[11] Libby A E, Wang H, Mittal R, et al. Lipoprotein lipase is an important modulator of lipid uptake and storage in hypothalamic neurons[J]. Biochemical & Biophysical Research Communications, 2015, 465(2):287-92.[12]张罕星,朱晓彤,束刚,周桂炫,高萍,高淑静,张常明,江青艳,陈瑶生. 猪肌内脂肪前体细胞与皮下脂肪前体细胞分化过程中基因差异表达分析[J]. 中国农业科学,2008,11:3760-3768.[13] Fan H, Wu D, Tian W, et al. Inhibitory effects of tannic acid on fatty acid synthase and 3T3-L1 preadipocyte.[J]. Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, 2013, 1831(7):1260-1266.[14] Innocenzi D, Alò P L, Balzani A, et al. Fatty acid synthase expression in melanoma.[J]. Journal of Cutaneous Pathology, 2003, 30(1):23-8.[15]张罕星,朱晓彤,束刚,高萍,高淑静,张常明,江青艳,陈瑶生. 猪脂肪前体细胞分化过程中聚脂相关基因的表达模式[J]. 动物学报,2007,01:143-150.[16] Cordonier E L, Jarecke S K, Hollinger F E, et al. Inhibition of acetyl-CoA carboxylases by soraphen A prevents lipid accumulation and adipocyte differentiation in 3T3-L1 cells.[J]. European Journal of Pharmacology, 2016, 780:202-208.[17] Schoiswohl G, Stefanovicracic M, Menke M N, et al. Impact of Reduced ATGL-Mediated Adipocyte Lipolysis on Obesity-Associated Insulin Resistance and Inflammation in Male Mice.[J]. Endocrinology, 2015, 156(10):3610-24.[18] Ahmadian M, Duncan R E, Varady K A, et al. Adipose overexpression of desnutrin promotes fatty acid use and attenuates diet-induced obesity.[J]. Diabetes, 2009, 58(4):855-866.[19]刘作华,杨飞云,孔路军,周晓容,辜玉红,王孝友. 日粮能量水平对生长育肥猪肌内脂肪含量以及脂肪酸合成酶和激素敏感脂酶mRNA表达的影响[J]. 畜牧兽医学报,2007,09:934-941.