精胺对雌鹅十二指肠多胺代谢关键基因表达及多胺含量的影响
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摘要
精胺是细胞增殖和分化的必需因子之一,不仅参与调节肠道上皮细胞增殖与凋亡、促进肠道发育,还参与调控肠道消化与吸收功能。为研究外源性精胺对四川白鹅(Anser cygnoides)雌鹅十二指肠多胺代谢的影响,按照鹅体重灌胃5和10 mg/kg精胺,应用qRT-PCR和高效液相色谱技术检测雌鹅十二指肠多胺代谢关键基因表达量及多胺含量。结果表明,5 mg/kg精胺灌胃组雌鹅十二指肠组织鸟氨酸脱羧酶抗酶抑制剂(ornithine decarboxylase antizyme inhibitor, AZIN) 1、鸟氨酸脱羧酶抗酶1 (ornithine decarboxylase antizyme 1, OAZ1)、精胺合成酶(spermine synthase, SPMS)、亚精胺/精胺N1-乙酰转移酶(spermidine/spermine-N1-acetyltransferase, SSAT)和乙酰多胺氧化酶(acetyl-polyamine oxidase, APAO)基因表达量均显著高于对照组(P<0.05);AZIN2基因表达量显著低于对照组(P<0.05);10 mg/kg精胺灌胃组雌鹅十二指肠AZIN1、OAZ1、鸟氨酸脱羧酶(ornithine decarboxylase, ODC)、亚精胺合成酶(spermidine synthase, SPDS)和SSAT的基因表达量均显著高于对照组(P<0.05)。5 mg/kg精胺灌胃组雌鹅十二指肠亚精胺(65.08 vs 41.84 mg/kg)和精胺(255.79 vs 218.84 mg/kg)含量显著高于对照组(P<0.05);10 mg/kg精胺灌胃组雌鹅十二指肠亚精胺(56.20 vs 41.84 mg/kg)含量显著高于对照组(P<0.05);而精胺(188.34 vs218.84 mg/kg)含量显著低于对照组(P<0.05)。综上所述,外源性精胺灌胃可通过介导多胺代谢关键酶基因表达而影响雌鹅十二指肠组织中亚精胺和精胺含量,对腐胺含量无显著影响。本研究揭示了外源性精胺对雌鹅十二指肠多胺代谢关键基因表达及多胺含量的影响,为精胺调控家禽消化与吸收的相关研究提供了基础数据,并可为开发新型功能性饲料提供新途径。
Spermine is one of the essential factors for cell proliferation and differentiation. The spermine in small intestine not only plays a role in regulating intestinal epithelial cell proliferation and apoptosis and promoting intestinal development, but also plays a role in regulating intestinal digestion and absorption. This experiment was conducted to study the effects of exogenous spermine on polyamine metabolism in female goose(Anser cygnoides) duodenum. Geese were fed spermine at 5 and 10 mg/kg(body weight). The expression of key genes related to polyamine metabolism and the content of polyamines in female geese duodenum were measured by qRT-PCR and high performance liquid chromatography. The gene expression levels of ornithine decarboxylase antizyme inhibitor(AZIN) 1, ornithine decarboxylase antizyme 1(OAZ1), spermine synthase(SPMS), spermidine/spermine-N1-acetyltransferase(SSAT) and acetyl-polyamine oxidase(APAO) in the duodenum of female geese treated with spermine at 5 mg/kg were significantly higher than those in the control group(P<0.05), while the gene expression level of AZIN2 significantly decreased(P<0.05). The gene expression levels of AZIN1, OAZ1, ornithine decarboxylase(ODC), spermidine synthase(SPDS) and SSAT in the duodenum of female geese treated with spermine at 10 mg/kg were significantly higher than those in the control group(P<0.05). The contents of spermidine(65.08 vs 41.84 mg/kg) and spermine(255.79 vs 218.84 mg/kg) in the duodenum of female geese treated with spermine at 5 mg/kg were significantly higher than those in the control group(P<0.05). Spermidine content(56.20 vs 41.84 mg/kg) in the duodenum of female geese treated with spermine at 10 mg/kg was significantly higher than that in the control group(P<0.05), while spermine content(188.34 vs 218.84 mg/kg) significantly decreased(P<0.05). In summary, exogenous spermine affects the contents of spermidine and spermine in female goose duodenum through regulating the expression of key genes related to polyamine metabolism, while it had no significant effect on putrescine content. This study revealed the effects of exogenous spermine on the expression of key genes of polyamine metabolism and polyamine content in duodenum of female geese, which provides basic data for the study of spermine regulation on digestion and absorption of poultry, and provides a new way for the development of new functional feedstuffs.
Spermine is one of the essential factors for cell proliferation and differentiation. The spermine in small intestine not only plays a role in regulating intestinal epithelial cell proliferation and apoptosis and promoting intestinal development, but also plays a role in regulating intestinal digestion and absorption. This experiment was conducted to study the effects of exogenous spermine on polyamine metabolism in female goose(Anser cygnoides) duodenum. Geese were fed spermine at 5 and 10 mg/kg(body weight). The expression of key genes related to polyamine metabolism and the content of polyamines in female geese duodenum were measured by qRT-PCR and high performance liquid chromatography. The gene expression levels of ornithine decarboxylase antizyme inhibitor(AZIN) 1, ornithine decarboxylase antizyme 1(OAZ1), spermine synthase(SPMS), spermidine/spermine-N1-acetyltransferase(SSAT) and acetyl-polyamine oxidase(APAO) in the duodenum of female geese treated with spermine at 5 mg/kg were significantly higher than those in the control group(P<0.05), while the gene expression level of AZIN2 significantly decreased(P<0.05). The gene expression levels of AZIN1, OAZ1, ornithine decarboxylase(ODC), spermidine synthase(SPDS) and SSAT in the duodenum of female geese treated with spermine at 10 mg/kg were significantly higher than those in the control group(P<0.05). The contents of spermidine(65.08 vs 41.84 mg/kg) and spermine(255.79 vs 218.84 mg/kg) in the duodenum of female geese treated with spermine at 5 mg/kg were significantly higher than those in the control group(P<0.05). Spermidine content(56.20 vs 41.84 mg/kg) in the duodenum of female geese treated with spermine at 10 mg/kg was significantly higher than that in the control group(P<0.05), while spermine content(188.34 vs 218.84 mg/kg) significantly decreased(P<0.05). In summary, exogenous spermine affects the contents of spermidine and spermine in female goose duodenum through regulating the expression of key genes related to polyamine metabolism, while it had no significant effect on putrescine content. This study revealed the effects of exogenous spermine on the expression of key genes of polyamine metabolism and polyamine content in duodenum of female geese, which provides basic data for the study of spermine regulation on digestion and absorption of poultry, and provides a new way for the development of new functional feedstuffs.
引文
陈咨余,姜冬梅,康波,等.2017.腹腔注射亚精胺对小鼠卵巢组织多胺含量及代谢相关基因表达的影响[J].华南农业大学学报,38(4):52-56.(Chen Z Y,Jiang D M,Kang B,et al.2017.Effects of intraperitoneal spermidine injection on polyamine content and metabolism-related gene expression in mouse ovary[J].Journal of South China Agricultural University,38(4):52-56.)
李恩中,陈燕芳,索世英,等.2010.外源精脒和精胺可诱导大鼠早期再生肝抗酶的表达[J].解剖学报,41(3):387-390.(Li E Z,Chen Y F,Suo S Y,et al.2010.Eexogenous spermidine and spermine induce the expression of antizyme in early regenerating rat liver[J].Acta Anatomica Sinica,41(3):387-390.)
易星,莫远亮,姜冬梅,等.2014.多胺的生物学功能及其调控机制[J].动物营养学报,26(2):348-352.(Yi X,Mo YL,Jiang D M,et al.2014.Biological functions of polyamine and its regulatory mechanisms[J].Chinese Journal of Animal Nutrition,26(2):348-352.)
易治鑫,姜冬梅,康波.2014.四川白鹅OAZ1和OAZ2基因组织差异表达的研究[J].华北农学报,29(1):50-54.(Yi Z X,Jiang D M,Kang B.2014.Study on differential expression of OAZ1 and OAZ2 gene in various tissues of the sichuan white goose[J].Acta Agriculturae Boreali-Sinica,29(1):50-54.)
赵宏涛,何余湧,陆伟.2015.外源精胺对断奶仔猪血液精胺含量、脏器发育和生产性能的影响[J].饲料工业,36(11):15-17.(Zhao H T,He Y Y,Lu W.2015.Effect of exogenous spermine on the level of spermine in blood,organ development and performance of weanerpiglets[J].Feed Industry,36(11):15-17.)
Cao W,Xu X,Jia G,et al.2018.Roles of spermine in modulating the antioxidant status and Nrf2 signalling molecules expression in the thymus and spleen of suckling piglets-new insight[J].Journal of Animal Physiology and Animal Nutrition,102(1):e183-e192.
Del Rio B,Redruello B,Linares D M,et al.2018.Spermine and spermidine are cytotoxic towards intestinal cell cultures,but are they a health hazard at concentrations found in foods?[J].Food Chemistry,269(2018):321-326.
Gevrekci A O.2017.The roles of polyamines in microorganisms[J].World Journal of Microbiology and Biotechnology,33(11):204.
Kahana C.2009.Antizyme and antizyme inhibitor,a regulatory tango[J].Cellular and Molecular Life Sciences,66(15):2479-2488.
Kang B,Jiang D,He H,et al.2017a.Effect of Oaz1 overexpression on goose ovarian granulosa cells[J].Amino Acids,49(6):1123-1132.
Kang B,Jiang D,Ma R,et al.2017b.OAZ1 knockdown enhances viability and inhibits ER and LHR transcriptions of granulosa cells in geese[J].PLOS One,12(3):e0175016.
Kurian L,Palanimurugan R,Godderz D,et al.2011.Polyamine sensing by nascent ornithine decarboxylase antizyme stimulates decoding of its m RNA[J].Nature,477(7365):490-494.
Li X,Coffino P.1992.Regulated degradation of ornithine decarboxylase requires interaction with the polyamine-inducible protein antizyme[J].Molecular and Cellular Biology,12(8):3556-3562.
Min S H,Simmen R C,Alhonen L,et al.2002.Altered levels of growth-related and novel gene transcripts in reproductive and other tissues of female mice overexpressing spermidine/spermine N1-acetyltransferase(SSAT)[J].Journal of Biological Chemistry,277(5):3647-3657.
Pegg A E.2014.The function of spermine[J].IUBMB Life,66(1):8-18.
Perez-Leal O,Barrero C A,Clarkson A B,et al.2012.Polyamine-regulated translation of spermidine/spermine-N1-acetyltransferase[J].Molecular and Cellular Biology,32(8):1453-1467.
Petros L M,Howard M T,Gesteland R F,et al.2005.Polyamine sensing during antizyme mRNA programmed frameshifting[J].Biochemical and Biophysical Research Communications,338(3):1478-1489.
Shappell N W,Fogel-Petrovic M F,Porter C W.1993.Regulation of spermidine/spermine N1-acetyltransferase by intracellular polyamine pools.Evidence for a functional role in polyamine homeostasis[J].FEBS Letters,321(2-3):179-183.
Snezhkina A V,Krasnov G S,Lipatova A V,et al.2016.The dysregulation of polyamine metabolism in colorectal cancer is associated with overexpression of c-Myc and C/EBPβrather than enterotoxigenic Bacteroides fragilis infection[J].Oxidative Medicine and Cellular Longevity,2016:2353560.
van Wettere W H,Willson N L,Pain S J,et al.2016.Effect of oral polyamine supplementation pre-weaning on piglet growth and intestinal characteristics[J].Animal,10(10):1655-1659.
Wallace H M.2009.The polyamines:Past,present and future[J].Essays in Biochemistry,46(2009):1-9.
Wu Y Y,Li T M,Zang L Q,et al.2018.Effects of berberine on tumor growth and intestinal permeability in HCT116tumor-bearing mice using polyamines as targets[J].Biomedicine Pharmacotherapy,107(2018):1447-1453.
李恩中,陈燕芳,索世英,等.2010.外源精脒和精胺可诱导大鼠早期再生肝抗酶的表达[J].解剖学报,41(3):387-390.(Li E Z,Chen Y F,Suo S Y,et al.2010.Eexogenous spermidine and spermine induce the expression of antizyme in early regenerating rat liver[J].Acta Anatomica Sinica,41(3):387-390.)
易星,莫远亮,姜冬梅,等.2014.多胺的生物学功能及其调控机制[J].动物营养学报,26(2):348-352.(Yi X,Mo YL,Jiang D M,et al.2014.Biological functions of polyamine and its regulatory mechanisms[J].Chinese Journal of Animal Nutrition,26(2):348-352.)
易治鑫,姜冬梅,康波.2014.四川白鹅OAZ1和OAZ2基因组织差异表达的研究[J].华北农学报,29(1):50-54.(Yi Z X,Jiang D M,Kang B.2014.Study on differential expression of OAZ1 and OAZ2 gene in various tissues of the sichuan white goose[J].Acta Agriculturae Boreali-Sinica,29(1):50-54.)
赵宏涛,何余湧,陆伟.2015.外源精胺对断奶仔猪血液精胺含量、脏器发育和生产性能的影响[J].饲料工业,36(11):15-17.(Zhao H T,He Y Y,Lu W.2015.Effect of exogenous spermine on the level of spermine in blood,organ development and performance of weanerpiglets[J].Feed Industry,36(11):15-17.)
Cao W,Xu X,Jia G,et al.2018.Roles of spermine in modulating the antioxidant status and Nrf2 signalling molecules expression in the thymus and spleen of suckling piglets-new insight[J].Journal of Animal Physiology and Animal Nutrition,102(1):e183-e192.
Del Rio B,Redruello B,Linares D M,et al.2018.Spermine and spermidine are cytotoxic towards intestinal cell cultures,but are they a health hazard at concentrations found in foods?[J].Food Chemistry,269(2018):321-326.
Gevrekci A O.2017.The roles of polyamines in microorganisms[J].World Journal of Microbiology and Biotechnology,33(11):204.
Kahana C.2009.Antizyme and antizyme inhibitor,a regulatory tango[J].Cellular and Molecular Life Sciences,66(15):2479-2488.
Kang B,Jiang D,He H,et al.2017a.Effect of Oaz1 overexpression on goose ovarian granulosa cells[J].Amino Acids,49(6):1123-1132.
Kang B,Jiang D,Ma R,et al.2017b.OAZ1 knockdown enhances viability and inhibits ER and LHR transcriptions of granulosa cells in geese[J].PLOS One,12(3):e0175016.
Kurian L,Palanimurugan R,Godderz D,et al.2011.Polyamine sensing by nascent ornithine decarboxylase antizyme stimulates decoding of its m RNA[J].Nature,477(7365):490-494.
Li X,Coffino P.1992.Regulated degradation of ornithine decarboxylase requires interaction with the polyamine-inducible protein antizyme[J].Molecular and Cellular Biology,12(8):3556-3562.
Min S H,Simmen R C,Alhonen L,et al.2002.Altered levels of growth-related and novel gene transcripts in reproductive and other tissues of female mice overexpressing spermidine/spermine N1-acetyltransferase(SSAT)[J].Journal of Biological Chemistry,277(5):3647-3657.
Pegg A E.2014.The function of spermine[J].IUBMB Life,66(1):8-18.
Perez-Leal O,Barrero C A,Clarkson A B,et al.2012.Polyamine-regulated translation of spermidine/spermine-N1-acetyltransferase[J].Molecular and Cellular Biology,32(8):1453-1467.
Petros L M,Howard M T,Gesteland R F,et al.2005.Polyamine sensing during antizyme mRNA programmed frameshifting[J].Biochemical and Biophysical Research Communications,338(3):1478-1489.
Shappell N W,Fogel-Petrovic M F,Porter C W.1993.Regulation of spermidine/spermine N1-acetyltransferase by intracellular polyamine pools.Evidence for a functional role in polyamine homeostasis[J].FEBS Letters,321(2-3):179-183.
Snezhkina A V,Krasnov G S,Lipatova A V,et al.2016.The dysregulation of polyamine metabolism in colorectal cancer is associated with overexpression of c-Myc and C/EBPβrather than enterotoxigenic Bacteroides fragilis infection[J].Oxidative Medicine and Cellular Longevity,2016:2353560.
van Wettere W H,Willson N L,Pain S J,et al.2016.Effect of oral polyamine supplementation pre-weaning on piglet growth and intestinal characteristics[J].Animal,10(10):1655-1659.
Wallace H M.2009.The polyamines:Past,present and future[J].Essays in Biochemistry,46(2009):1-9.
Wu Y Y,Li T M,Zang L Q,et al.2018.Effects of berberine on tumor growth and intestinal permeability in HCT116tumor-bearing mice using polyamines as targets[J].Biomedicine Pharmacotherapy,107(2018):1447-1453.
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