雌激素对奶牛乳腺上皮细胞中酪蛋白和甘油三酯表达的影响
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摘要
为探讨雌激素对乳腺上皮细胞中酪蛋白及甘油三酯表达的影响,试验于2018年2月至2018年4月在新疆肉乳用草食动物营养实验室进行,实验选取扩增生长状态良好的乳腺上皮细胞,添加0、50、100、200μmol/l雌激素孵育乳腺上皮细胞后,分别在12、24、48、72 h后检测各组细胞酪蛋白及甘油三酯表达的情况。结果显示,在12 h时,添加50μmol/l雌激素组CSN1的表达量极显著高于对照组(P<0.01),添加50μmol/l雌激素组TG表达量极显著高于对照组(P<0.01);50μmol/l雌激素添加组CSN1表达量显著高于200μmol/l雌激素添加组(P<0.05),且极显著高于100μmol/l雌激素添加组(P<0.01),100μmol/l雌激素添加组CSN2表达量显著高于200μmol/l雌激素添加组(P<0.05),50μmol/l雌激素添加组TG表达量极显著高于100、200μmol/l雌激素添加组(P<0.01);添加50μmol/l雌激素时,12、48、72 h的CSN1表达量显著高于24 h(P<0.05),在72 hCSN2表达量显著高于其他各时间点(P<0.05),24、72 h时TG表达量显著高于12、48 h(P<0.05)。结果发现,添加雌激素可以促进乳腺上皮细胞中CSN1、CSN2及TG的表达。200μmol/l在一定条件下促进CSN1的表达效果最好;100、200μmol/l雌激素添加组均能够促进CSN2的表达,且100μmol/l促进CSN2的表达效果最好,尤其是在12、24 h;50、200μmol/l雌激素添加组可以促进TG的表达,但100μmol/l雌激素添加组能够抑制TG的表达。
To investigate the effect of estrogen on the expression of casein and triglyceride in bovine mammary epithelial cells. The experiment was carried out in Xinjiang Meat and Milk Herbivorous Animal Nutrition Laboratory from February 2018 to April 2018. The bovine mammary epithelial cells with good proliferative status were selected and incubated with 0, 50, 100, 200 μmol/l estrogen. The expression of casein and triglyceride in each group was detected after 12, 24, 48 h and 72 h respectively.The results showed that at 12 hours, the expression of CSN1 in 50 μmol/l estrogen supplementation group was extremely significant higher than that in the control group(P<0.01). The expression of TG in 50 μmol/l estrogen group was significantly higher than that in control group(P<0.05). The expression of CSN1 in 50 μmol/l estrogen supplementation group was significantly higher than that in200 μmol/l estrogen supplementation group(P<0.05), and extremely significant higher than that in 100 μmol/l estrogen supplementation group(P<0.01). The expression of CSN2 in 100 μmol/l estrogen supplementation group was significantly higher than that in 200 μmol/l estrogen supplementation group(P<0.05). The expression of TG in 50 μmol/l estrogen supplementation group was extremely significant higher than that in 100 μmol/l and 200 μmol/l estrogen supplementation group(P<0.01). When 50 μmol/l estrogen was added, the expression of CSN1 at 12, 48 h and 72 h was significantly higher than that at 24 h(P<0.05). The expression of CSN2 at 72 h was significantly higher than that at other time points(P<0.05), and the expression of TG at 24 h and 72 h was significantly higher than that at 12 h and 48 h(P<0.05). Estrogen could promote the expression of CSN1, CSN2 and TG in bovine mammary epithelial cells. 200 μmol/l could promote the expression of CSN1 best under certain conditions; 100 and 200 μmol/l estrogen supplementation group could promote the expression of CSN2, and 100 μmol/l could promote the expression of CSN2 best, especially at 12 h and24 h; 50 μmol/l and 200 μmol/l estrogen supplementation group could promote the expression of TG,but 100 μmol/l estrogen supplementation group could inhibit the expression of TG.
To investigate the effect of estrogen on the expression of casein and triglyceride in bovine mammary epithelial cells. The experiment was carried out in Xinjiang Meat and Milk Herbivorous Animal Nutrition Laboratory from February 2018 to April 2018. The bovine mammary epithelial cells with good proliferative status were selected and incubated with 0, 50, 100, 200 μmol/l estrogen. The expression of casein and triglyceride in each group was detected after 12, 24, 48 h and 72 h respectively.The results showed that at 12 hours, the expression of CSN1 in 50 μmol/l estrogen supplementation group was extremely significant higher than that in the control group(P<0.01). The expression of TG in 50 μmol/l estrogen group was significantly higher than that in control group(P<0.05). The expression of CSN1 in 50 μmol/l estrogen supplementation group was significantly higher than that in200 μmol/l estrogen supplementation group(P<0.05), and extremely significant higher than that in 100 μmol/l estrogen supplementation group(P<0.01). The expression of CSN2 in 100 μmol/l estrogen supplementation group was significantly higher than that in 200 μmol/l estrogen supplementation group(P<0.05). The expression of TG in 50 μmol/l estrogen supplementation group was extremely significant higher than that in 100 μmol/l and 200 μmol/l estrogen supplementation group(P<0.01). When 50 μmol/l estrogen was added, the expression of CSN1 at 12, 48 h and 72 h was significantly higher than that at 24 h(P<0.05). The expression of CSN2 at 72 h was significantly higher than that at other time points(P<0.05), and the expression of TG at 24 h and 72 h was significantly higher than that at 12 h and 48 h(P<0.05). Estrogen could promote the expression of CSN1, CSN2 and TG in bovine mammary epithelial cells. 200 μmol/l could promote the expression of CSN1 best under certain conditions; 100 and 200 μmol/l estrogen supplementation group could promote the expression of CSN2, and 100 μmol/l could promote the expression of CSN2 best, especially at 12 h and24 h; 50 μmol/l and 200 μmol/l estrogen supplementation group could promote the expression of TG,but 100 μmol/l estrogen supplementation group could inhibit the expression of TG.
引文
[1]赵国丽,宫艳斌,韩元,等.激素和生长因子调控奶牛乳腺发育的研究进展[J].中国奶牛,2011(6):25-30.
[2]李冬辉.催乳素、雌激素和胰岛素对奶牛乳腺上皮细胞中瘦素及其受体的影响[D].哈尔滨:东北农业大学,2012.
[3]姚志兰,崔平福,宗佳丽,等.奶牛泌乳启动调控激素的研究进展[J].畜牧与兽医,2018,50(8):129-132.
[4]杨慧敏,谷文举.大豆黄酮对奶牛泌乳及相关激素的影响[J].今日畜牧兽医,2007(7):9-10.
[5]郝振荣,朱志宁,郭玉琴,等.大豆黄酮对产奶量和牛奶品质的影响[J].北京农学院学报,2009,24(4):26-27,38.
[6]赵传毅,姜宁,杨文忠,等.饲喂不同水平大豆异黄酮对泌乳中期奶牛生产性能的影响[J].黑龙江畜牧兽医,2011(17):68-69.
[7]曹艳红,罗军,张蒙,等.体外培养奶山羊乳腺上皮细胞的形态学观察[J].中国兽医杂志,2008,44(5):20-21.
[8] Warzych E, Cieslak A, Pawlak P, et al. Maternal nutrition affects the comPosition of follicular fluid and transcript content in gilt oocytes[J]. Veterinarni Medicina, 2011,56:156-167.
[9] Tan D, Johnson D A, Wu W, et al. Unmodified prolactin(PRL)and S179D PRL-initiated bioluminescence resonance energy transfer between homo-and hetero-pairs of long and short human PRL receptors in living human cells[J]. Molecular Endocrinology,2005, 19(5):1291-1303.
[10]杨建英,张勇法.大豆黄酮对奶牛产奶量和乳中常规成分的影响[J].饲料研究,2005(6):30-31.
[11]艾晓杰,吴晓林,朱勇琪,等.大豆黄酮对荷斯坦牛乳中蛋白质和脂肪含量的影响[J].中国奶牛,2005(6):19-20.
[12] Albenzio M, Santillo A, Caroprese M, et al. Effect of stage of lactation on the immune competence of goat mammary gland[J].Journal of Dairy Science, 2016, 99(5):3889-3895.
[13] Muldoon T G. Interplay between Estradiol and Prolactin in the Regulation of Steroid Hormone Receptor Levels, Nature and Functionality in Normal Mouse Mammary Tissue[J]. Endocrinology, 1981,109(5):1339-1346.
[14] Berryhill G E, Trott J F, Hovey R C. Mammary gland development-It's not just about estrogen[J]. Journal of dairy science,2016,99(1):875-883.
[15]卢志勇,梁代华,杨运玲,等.大豆异黄酮对奶牛乳腺上皮细胞泌乳性能及抗氧化能力的影响[J].饲料与畜牧,2013(2):25-29.
[16]赵传毅,姜宁,杨文忠,等.饲喂不同水平大豆异黄酮对泌乳中期奶牛生产性能的影响[J].黑龙江畜牧兽医,2011(17):68-69.
[17]朱河水,王艳玲,杨国宇,等.大豆黄酮对奶牛相关产奶性能的影响[J].华北农学报,2006,21(6):127-129.
[18] Backwell F R, Bequette B J, Wilson D, et al. Evidence for the utilization of PePtides for milk Protein synthesis in the lactating dairy goat in vivo[J].American Journal of Physiology, 1996, 271(2):955-960.
[19] Barash I, FaermanA, Richenstein M, et al. In vivo and in vitro expression of human serum albumin genomic sequences in mammary epithelial cells with beta-lactoglobu-lin and whey acidic Protein Promoters[J]. Mol. Reprod. Dev., 1999,52(3):241-252.
[20] Scully K M, Gleiberman A S, Lindzey J, et al. Role of estrogen receptor-alpha in the anterior pituitary gland[J]. Molecular Endocrinology, 1997, 11(6):674-681.
[21]程蕾,王定发,刘晓华,等.大豆异黄酮对奶牛产奶量及乳成分的影响[J].饲料博览, 2009(4):5-7.
[22]李冬.METTL3对奶牛乳腺上皮细胞乳蛋白和乳脂肪合成的调控及其作用机理[D].哈尔滨:东北农业大学,2017.
[23]李艳,徐欣然,徐春奎,等.催乳激素和赖氨酸对奶牛乳腺上皮细胞分泌乳蛋白的影响[J].上海畜牧兽医通讯, 2018(5):2-7.
[24] Ismail P M, Amato P, Soyal S M, et al. Progesterone involvement in breast development and Tumorigenesis-as revealed by progesterone receptor"knockout"and"knockin"mouse models[J]. Steroids, 2003, 68(10/13):779-787.
[2]李冬辉.催乳素、雌激素和胰岛素对奶牛乳腺上皮细胞中瘦素及其受体的影响[D].哈尔滨:东北农业大学,2012.
[3]姚志兰,崔平福,宗佳丽,等.奶牛泌乳启动调控激素的研究进展[J].畜牧与兽医,2018,50(8):129-132.
[4]杨慧敏,谷文举.大豆黄酮对奶牛泌乳及相关激素的影响[J].今日畜牧兽医,2007(7):9-10.
[5]郝振荣,朱志宁,郭玉琴,等.大豆黄酮对产奶量和牛奶品质的影响[J].北京农学院学报,2009,24(4):26-27,38.
[6]赵传毅,姜宁,杨文忠,等.饲喂不同水平大豆异黄酮对泌乳中期奶牛生产性能的影响[J].黑龙江畜牧兽医,2011(17):68-69.
[7]曹艳红,罗军,张蒙,等.体外培养奶山羊乳腺上皮细胞的形态学观察[J].中国兽医杂志,2008,44(5):20-21.
[8] Warzych E, Cieslak A, Pawlak P, et al. Maternal nutrition affects the comPosition of follicular fluid and transcript content in gilt oocytes[J]. Veterinarni Medicina, 2011,56:156-167.
[9] Tan D, Johnson D A, Wu W, et al. Unmodified prolactin(PRL)and S179D PRL-initiated bioluminescence resonance energy transfer between homo-and hetero-pairs of long and short human PRL receptors in living human cells[J]. Molecular Endocrinology,2005, 19(5):1291-1303.
[10]杨建英,张勇法.大豆黄酮对奶牛产奶量和乳中常规成分的影响[J].饲料研究,2005(6):30-31.
[11]艾晓杰,吴晓林,朱勇琪,等.大豆黄酮对荷斯坦牛乳中蛋白质和脂肪含量的影响[J].中国奶牛,2005(6):19-20.
[12] Albenzio M, Santillo A, Caroprese M, et al. Effect of stage of lactation on the immune competence of goat mammary gland[J].Journal of Dairy Science, 2016, 99(5):3889-3895.
[13] Muldoon T G. Interplay between Estradiol and Prolactin in the Regulation of Steroid Hormone Receptor Levels, Nature and Functionality in Normal Mouse Mammary Tissue[J]. Endocrinology, 1981,109(5):1339-1346.
[14] Berryhill G E, Trott J F, Hovey R C. Mammary gland development-It's not just about estrogen[J]. Journal of dairy science,2016,99(1):875-883.
[15]卢志勇,梁代华,杨运玲,等.大豆异黄酮对奶牛乳腺上皮细胞泌乳性能及抗氧化能力的影响[J].饲料与畜牧,2013(2):25-29.
[16]赵传毅,姜宁,杨文忠,等.饲喂不同水平大豆异黄酮对泌乳中期奶牛生产性能的影响[J].黑龙江畜牧兽医,2011(17):68-69.
[17]朱河水,王艳玲,杨国宇,等.大豆黄酮对奶牛相关产奶性能的影响[J].华北农学报,2006,21(6):127-129.
[18] Backwell F R, Bequette B J, Wilson D, et al. Evidence for the utilization of PePtides for milk Protein synthesis in the lactating dairy goat in vivo[J].American Journal of Physiology, 1996, 271(2):955-960.
[19] Barash I, FaermanA, Richenstein M, et al. In vivo and in vitro expression of human serum albumin genomic sequences in mammary epithelial cells with beta-lactoglobu-lin and whey acidic Protein Promoters[J]. Mol. Reprod. Dev., 1999,52(3):241-252.
[20] Scully K M, Gleiberman A S, Lindzey J, et al. Role of estrogen receptor-alpha in the anterior pituitary gland[J]. Molecular Endocrinology, 1997, 11(6):674-681.
[21]程蕾,王定发,刘晓华,等.大豆异黄酮对奶牛产奶量及乳成分的影响[J].饲料博览, 2009(4):5-7.
[22]李冬.METTL3对奶牛乳腺上皮细胞乳蛋白和乳脂肪合成的调控及其作用机理[D].哈尔滨:东北农业大学,2017.
[23]李艳,徐欣然,徐春奎,等.催乳激素和赖氨酸对奶牛乳腺上皮细胞分泌乳蛋白的影响[J].上海畜牧兽医通讯, 2018(5):2-7.
[24] Ismail P M, Amato P, Soyal S M, et al. Progesterone involvement in breast development and Tumorigenesis-as revealed by progesterone receptor"knockout"and"knockin"mouse models[J]. Steroids, 2003, 68(10/13):779-787.