TNC基因在牛肌肉发育和前体脂肪细胞诱导分化过程中的表达研究
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摘要
【目的】分析TNC基因在牛背最长肌组织发育和前体脂肪细胞诱导分化过程中的表达变化趋势及其与脂肪沉积之间的关系。【方法】利用qRT-PCR技术分析了3、6、12、18、24和30月龄南阳牛背最长肌组织中TNC基因的表达变化,利用索氏抽提法测定了24头牛背最长肌组织中的脂肪含量。分离了牛前体脂肪细胞,通过在培养基中添加2μmol/L柠檬苦素干扰细胞的分化,利用qRT-PCR技术分析了正常诱导分化和柠檬苦素干扰后的诱导分化过程中TNC基因的表达变化。【结果】TNC基因在18月龄之后的牛背最长肌组织中的表达水平逐渐升高。根据肌肉组织中脂肪含量的高低分为高肌内脂肪含量组和低肌内脂肪含量组,发现TNC基因的表达水平在这2组之间没有显著差异。TNC基因在牛前体脂肪细胞诱导分化开始后的第6小时表达量急剧升高,之后开始下降;在诱导分化的后期,当细胞质中有明显的脂质液滴积累时,TNC基因的表达水平再次升高。在诱导分化培养基中添加柠檬苦素能够抑制细胞的分化和脂质的积累,同时也导致TNC基因表达量显著下降。【结论】TNC基因参与了脂肪细胞的分化和脂质沉积的调控过程,该基因表达量上调有利于细胞质中脂质液滴的积累。
[Purpose]In order to analyze the expression pattern of TNC gene during the development of longissimus dorsi muscle and the differentiation of preadipocyte, and the relationship between TNC gene expression level with fat deposition in beef cattle. [Methods]The expression of TNC gene was analyzed by qRT-PCR technology in longissimus dorsi muscle tissues of 3, 6, 12, 18, 24 and30 months old Nanyang cattle. The fat content in longissimus dorsi muscle of 24 cattle was determined by Soxhlet extraction. The preadipocyte of cattle was separated and induced differentiation with normal medium or medium with 2 μmol/L limonin, and the expression of TNC gene was analyzed by qRT-PCR technology during the differentiation of preadipocyte in control and treatment groups respectively. [ Results] The expression of the TNC gene increased gradually in longissimus dorsi muscle after 18 month old in cattle. The muscle tissues were divided into high intramuscular fat group and low intramuscular fat group according to the fat content, and the expression levels showed no significant difference between these two groups. The expression level of TNC gene increased sharply at the 6 th hour of the differentiation, then decreased slowly, but increased again at the end of differentiation with the deposition of lipid droplet in cytoplasm. The differentiation of preadipocytes and lipid accumulation in the cytoplasm could be inhibited by limonin in medium, which also induced a significant decrease of TNC gene expression. [Conclusion]TNC gene is involved in the regulation of differentiation and lipid deposition in cultivated preadipocytes and fat tissues, and the up-regulated expression of TNC gene would benefit the lipid accumulation in cytoplasm.
[Purpose]In order to analyze the expression pattern of TNC gene during the development of longissimus dorsi muscle and the differentiation of preadipocyte, and the relationship between TNC gene expression level with fat deposition in beef cattle. [Methods]The expression of TNC gene was analyzed by qRT-PCR technology in longissimus dorsi muscle tissues of 3, 6, 12, 18, 24 and30 months old Nanyang cattle. The fat content in longissimus dorsi muscle of 24 cattle was determined by Soxhlet extraction. The preadipocyte of cattle was separated and induced differentiation with normal medium or medium with 2 μmol/L limonin, and the expression of TNC gene was analyzed by qRT-PCR technology during the differentiation of preadipocyte in control and treatment groups respectively. [ Results] The expression of the TNC gene increased gradually in longissimus dorsi muscle after 18 month old in cattle. The muscle tissues were divided into high intramuscular fat group and low intramuscular fat group according to the fat content, and the expression levels showed no significant difference between these two groups. The expression level of TNC gene increased sharply at the 6 th hour of the differentiation, then decreased slowly, but increased again at the end of differentiation with the deposition of lipid droplet in cytoplasm. The differentiation of preadipocytes and lipid accumulation in the cytoplasm could be inhibited by limonin in medium, which also induced a significant decrease of TNC gene expression. [Conclusion]TNC gene is involved in the regulation of differentiation and lipid deposition in cultivated preadipocytes and fat tissues, and the up-regulated expression of TNC gene would benefit the lipid accumulation in cytoplasm.
引文
[1]FRY C S,KIRBY T J,KOSMAC E,et al.Myogenic progenitor cells control extracellular matrix production by fibroblasts during skeletal muscle hypertrophy[J].Cell Stem Cell,2017,5,20(1):56.DOI:10.1016/j.stem.2016.09.010.
[2]C?TéJ A,GUéNARD F,LESSARD J,et al.Temporal changes in gene expression profile during mature adipocyte dedifferentiation[J].International Journal of Genomics,2017,2017:5149362.DOI:10.1155/2017/5149362.
[3]RAJPRASAD L,BRENDA J R,CHRISTOPHER M S,et al.Extracellular matrix motion and early morphogenesis[J].Development,2016,15,143(12):2056.DOI:10.1242/dev.127886.
[4]ADAMS J C,CHIQUET-EHRISMANN R,TUCKER RP.The evolution of tenascins and fibronectins[J].Cell Adhesion&Migration,2014,30:1.DOI:10.4161/19336918.2014.970030.
[5]MURPHY-ULLRICH J E,SAGE E H.Revisiting the matricellular concept[J].Matrix Biology,2014,37:1.DOI:10.1016/j.matbio.2014.07.005.
[6]ERICKSON H P.Tenascin-C,tenascin-R and tenascinX:a family of talented proteins in search of functions[J].Current Opinion in Cell Biology,1993,5:869.
[7]GHERZI R,CARNEMOLLA B,SIRI A,et al.Human tenascin gene.Structure of the 5’-region,identification,and characterization of the transcription regulatory sequences[J].Journal of Biological Chemistry,1995,270:3429.
[8]祁艳霞,张小辉,王玉琴,等.牛肌肉组织中胶原和ADAMTS2基因表达与肉质的相关性分析[J].云南农业大学学报(自然科学),2014,29(2):173.DOI:10.3969/j.issn.1004-390X(n).2014.02.005.
[9]祁艳霞,张小辉,王玉琴,等.基质金属蛋白酶2,3,13基因表达水平与牛肉质性状的相关性分析[J].云南农业大学学报(自然科学),2016,31(1):62.DOI:10.16211/j.issn.1004-390X(n).2016.01.010.
[10]徐铁山,张小辉,刘佩,等.柠檬苦素对牛前体脂肪细胞分化过程中核纤层蛋白基因表达的影响[J].黑龙江畜牧兽医,2017(3):10.DOI:10.13881/j.cnki.hljxmsy.2017.0195.
[11]KABLAN A,SAUNDERS R A,SZKUDLAREK-MIKHO M,et al.Prieurianin causes weight loss in dietinduced obese mice and inhibits adipogenesis in cultured preadipocytes[J].Journal of Diabetes&Metabolism,2010,1(101):1.
[12]曲桂娟,秦贵信.肉牛肌内脂肪的研究进展[J].中国畜牧兽医,2010,37(6):221.
[13]TUCKER R P,CHIQUET-EHRISMANN R.The regulation of tenascin expression by tissue microenvironments[J].Biochimica et Biophysica Acta-Molecular Basis of Disease,2009,1793:888.DOI:10.1016/j.bbamcr.2008.12.012.
[14]IMANAKA-YOSHIDA K,AOKI H.Tenascin-C and mechanotransduction in the development and diseases of cardiovascular system[J].Frontiers in Physiology,2014,5:283.DOI:10.3389/fphys.2014.00283.
[15]BOHANNON-STEWART A,GARY K,BONIFACE K,et al.Expression of potential regulatory genes in abdominal adipose tissue of broiler chickens during early development[J].Genetics Research International,2014:318304.DOI:10.1155/2014/318304.
[16]ZHANG X H,QI Y X,GAO X,et al.Expression of ADAMTS4 and ADAMTS5 in longissimus dorsi muscle related to meat tenderness in Nanyang cattle[J].Genetics and Molecular Research,2013,12(4):4639.DOI:10.4238/2013.October.18.2.
[17]QI Y X,ZHANG X H,WANG Y Q,et al.Expression of MMP-1,-2,and-8 in longissimus dorsi muscle and their relationship with meat quality traits in cattle[J].Genetics and Molecular Research,2016,15(1):1.DOI:10.4238/gmr.15017593.
[18]MARIMAN E C M,WANG P.Adipocyte extracellular matrix composition,dynamics and role in obesity[J].Cellular and Molecular Life Sciences,2010,67:1277.DOI:10.1007/s00018-010-0263-4.
[19]CHIOVARO F,CHIQUET-EHRISMANN R,CHI-QUET M.Transcriptional regulation of tenascin genes[J].Cell Adhesion&Migration,2015,9(1/2):34.DOI:10.1080/19336918.2015.1008333.
[20]THOH M,BABAJAN B,RAGHAVENDRA P B,et al.Azadirachtin interacts with retinoic acid receptors and inhibits retinoic acid-mediated biological responses[J].Journal of Biological Chemistry,2011,286(6):4690.DOI:10.1074/jbc.M110.169334.
[21]CHUNG C Y,MURPHY-ULLRICH J E,ERICKSON HP.Mitogenesis,cell migration,and loss of focal adhesions induced by tenascin-C interacting with its cell surface receptor,annexinⅡ[J].Molecular Biology of the Cell,1996,7:883.
[22]KIM S M,MATTHIAS C,RICHARD P T,et al.Tenascin-C at a glance[J].Journal of Cell Science,2016,129(23):4321.DOI:10.1242/jcs.190546.
[2]C?TéJ A,GUéNARD F,LESSARD J,et al.Temporal changes in gene expression profile during mature adipocyte dedifferentiation[J].International Journal of Genomics,2017,2017:5149362.DOI:10.1155/2017/5149362.
[3]RAJPRASAD L,BRENDA J R,CHRISTOPHER M S,et al.Extracellular matrix motion and early morphogenesis[J].Development,2016,15,143(12):2056.DOI:10.1242/dev.127886.
[4]ADAMS J C,CHIQUET-EHRISMANN R,TUCKER RP.The evolution of tenascins and fibronectins[J].Cell Adhesion&Migration,2014,30:1.DOI:10.4161/19336918.2014.970030.
[5]MURPHY-ULLRICH J E,SAGE E H.Revisiting the matricellular concept[J].Matrix Biology,2014,37:1.DOI:10.1016/j.matbio.2014.07.005.
[6]ERICKSON H P.Tenascin-C,tenascin-R and tenascinX:a family of talented proteins in search of functions[J].Current Opinion in Cell Biology,1993,5:869.
[7]GHERZI R,CARNEMOLLA B,SIRI A,et al.Human tenascin gene.Structure of the 5’-region,identification,and characterization of the transcription regulatory sequences[J].Journal of Biological Chemistry,1995,270:3429.
[8]祁艳霞,张小辉,王玉琴,等.牛肌肉组织中胶原和ADAMTS2基因表达与肉质的相关性分析[J].云南农业大学学报(自然科学),2014,29(2):173.DOI:10.3969/j.issn.1004-390X(n).2014.02.005.
[9]祁艳霞,张小辉,王玉琴,等.基质金属蛋白酶2,3,13基因表达水平与牛肉质性状的相关性分析[J].云南农业大学学报(自然科学),2016,31(1):62.DOI:10.16211/j.issn.1004-390X(n).2016.01.010.
[10]徐铁山,张小辉,刘佩,等.柠檬苦素对牛前体脂肪细胞分化过程中核纤层蛋白基因表达的影响[J].黑龙江畜牧兽医,2017(3):10.DOI:10.13881/j.cnki.hljxmsy.2017.0195.
[11]KABLAN A,SAUNDERS R A,SZKUDLAREK-MIKHO M,et al.Prieurianin causes weight loss in dietinduced obese mice and inhibits adipogenesis in cultured preadipocytes[J].Journal of Diabetes&Metabolism,2010,1(101):1.
[12]曲桂娟,秦贵信.肉牛肌内脂肪的研究进展[J].中国畜牧兽医,2010,37(6):221.
[13]TUCKER R P,CHIQUET-EHRISMANN R.The regulation of tenascin expression by tissue microenvironments[J].Biochimica et Biophysica Acta-Molecular Basis of Disease,2009,1793:888.DOI:10.1016/j.bbamcr.2008.12.012.
[14]IMANAKA-YOSHIDA K,AOKI H.Tenascin-C and mechanotransduction in the development and diseases of cardiovascular system[J].Frontiers in Physiology,2014,5:283.DOI:10.3389/fphys.2014.00283.
[15]BOHANNON-STEWART A,GARY K,BONIFACE K,et al.Expression of potential regulatory genes in abdominal adipose tissue of broiler chickens during early development[J].Genetics Research International,2014:318304.DOI:10.1155/2014/318304.
[16]ZHANG X H,QI Y X,GAO X,et al.Expression of ADAMTS4 and ADAMTS5 in longissimus dorsi muscle related to meat tenderness in Nanyang cattle[J].Genetics and Molecular Research,2013,12(4):4639.DOI:10.4238/2013.October.18.2.
[17]QI Y X,ZHANG X H,WANG Y Q,et al.Expression of MMP-1,-2,and-8 in longissimus dorsi muscle and their relationship with meat quality traits in cattle[J].Genetics and Molecular Research,2016,15(1):1.DOI:10.4238/gmr.15017593.
[18]MARIMAN E C M,WANG P.Adipocyte extracellular matrix composition,dynamics and role in obesity[J].Cellular and Molecular Life Sciences,2010,67:1277.DOI:10.1007/s00018-010-0263-4.
[19]CHIOVARO F,CHIQUET-EHRISMANN R,CHI-QUET M.Transcriptional regulation of tenascin genes[J].Cell Adhesion&Migration,2015,9(1/2):34.DOI:10.1080/19336918.2015.1008333.
[20]THOH M,BABAJAN B,RAGHAVENDRA P B,et al.Azadirachtin interacts with retinoic acid receptors and inhibits retinoic acid-mediated biological responses[J].Journal of Biological Chemistry,2011,286(6):4690.DOI:10.1074/jbc.M110.169334.
[21]CHUNG C Y,MURPHY-ULLRICH J E,ERICKSON HP.Mitogenesis,cell migration,and loss of focal adhesions induced by tenascin-C interacting with its cell surface receptor,annexinⅡ[J].Molecular Biology of the Cell,1996,7:883.
[22]KIM S M,MATTHIAS C,RICHARD P T,et al.Tenascin-C at a glance[J].Journal of Cell Science,2016,129(23):4321.DOI:10.1242/jcs.190546.