胰岛素对体外培养奶牛乳腺上皮细胞乳蛋白、乳脂肪合成相关基因mRNA表达的影响

doi:10.11843/j.issn.0366-6964.2013.05.007

Abstract

Abstract:

This experiment was conducted to study the effects of insulin on mRNA expression of genes related to milk protein and fat synthesis in bovine mammary cells cultured in vitro. Mammary epithelial cells were cultured for 24 h with the following hormone treatments: no hormones (NH, control), 50 ng·mL^-1 hydrocortisone + 200 ng·mL^-1 prolactin (FP) or 100 ng·mL^-1 insulin + 50 ng·mL^-1 hydrocortisone + 200 ng·mL^-1 prolactin (IFP). The relative expression of the related genes of milk protein and fat synthesis were measured by real-time PCR. The results showed that the mRNA relative abundance of β-casein (CSN2), κ-casein (CSN3), Acetyl-CoA carboxylase (ACACA), fatty acid synthase (FASN) and sterol regulatory element pinding protein1 (SREBP1) in IFP group were significantly up-regulated(P<0.05). In addition, signal transducers and activators of transduction 5(STAT5B) and ETS-related transcription factor Elf-5 (ELF5) in JAK2-STAT5 signal pathway, as well as Phosphoinositide-3-kinase (PI3K), protein kinase B(AKT1) and eukaryotic translation initiation factor 4E(EIF4E) in PI3K/Akt/mTOR siganal pathway in IFP group were significantly up-regulated (P<0.05), while the IFP hormone combinations had no effect on TSC1, TSC2 or RHEB transcription in AMPK singal pathway. The data demonstrated that insulin induced the mRNA expression of genes related to milk protein synthesis through JAK2-STAT5 and PI3K/Akt/mTOR signal pathway and the mRNA expression of genes related to milk fat synthesis through PI3K/Akt/mTOR and SREBP signal pathway in bovine mammary epithelial cells cultured in vitro, which indicated insulin, hydrocortisone and prolactin regulate the mRNA expression of genes related to milk protein and fat synthes together.

CLC Number:

S823.9⁺1.2

WANG Hao-yu, QIN Tong, HAO Hai-sheng, DU Wei-hua, ZHAO Xue-ming, ZHU Hua-bin. Effects of Insulin on mRNA Expression of Genes Related to Milk Protein and Fat Synthesis in Bovine Mammary Epithelial Cells Cultured in vitro[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(5): 710-718.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.xmsyxb.com/EN/10.11843/j.issn.0366-6964.2013.05.007

https://www.xmsyxb.com/EN/Y2013/V44/I5/710

References

ERDMANN B, BRETER H. Irregular distribution of mammary-derived growth inhibitor in the bovine mammary epithelium［J］. Cell Tissue Res, 1993, 272(2): 383-389.

ACCORNERO P, MARTIGNANI E, MIRETTI S, et al. Epidermal growth factor and hepatocyte growth factor receptors collaborate to induce multiple biological responses in bovine mammary epithelial cells［J］. J Dairy Sci, 2009, 92(8): 3667-3675.

佟慧丽，高学军，李庆章，等.胰岛素、催乳素对奶山羊乳腺上皮细胞泌乳功能的影响［J］.畜牧兽医学报，2008，39(6)：721-725.

FREEMAN M E, KANYICSKA B, LERANT A, et al. Prolactin: structure, function, and regulation of secretion［J］. Physiol Rev, 2000, 80(4): 1523-1631.

LEE H J, ORMANDY C J. Interplay between progesterone and prolactin in mammary development and implications for breast cancer［J］. Mol Cell Endocrinol, 2012, 357(1-2): 101-107.

李敏.新型糖皮质激素对肥大细胞脱颗粒的快速作用及其机制研究［D］.上海: 第二军医大学，2010.

李真，李庆章.奶山羊乳腺发育过程中生长激素、胰岛素及其受体的变化规律研究［J］.中国农业科学，2010，43(8)：1730-1737.

SHEEHY P A, NICHOLAS K R, WYNN P C. An investigation of the role of insulin in bovine milk protein gene expression in mammary explant culture［J］. Asian-Aus J Anim Sci, 2000, Supplement (13): 272-275.

MENZIES K K, LEFEVRE C, MACMILLAN K L, et al. Insulin regulates milk protein synthesis at multiple levels in the bovine mammary gland［J］. Funct Integrat Genom, 2009, 9(2): 197-217.

BIONAZ M, LOOR J J. Gene networks driving bovine mammary protein synthesis during the lactation cycle［J］. Bioinform Biol Insights, 2011, 5: 83-98.

HORTON J D, GOLDSTEIN J L, BROWN M S. SREBPs: activators of the complete program of cholesterol and fatty acid synthesis in the liver［J］. J Clin Invest, 2002, 109(9): 1125-1131.

WANG X, PROUD C G. The mTOR pathway in the control of protein synthesis［J］. Physiology (Bethesda), 2006, 21: 362-369.

PETERSON T R, SENGUPTA S S, HARRIS T E, et al. mTOR complex 1 regulates lipin 1 localization to control the SREBP pathway［J］. Cell, 2011, 146(3): 408-420.

张霞，孙琳琳，钟殿胜. LKB1-AMPK-mTOR信号传导通路在肿瘤中的研究进展［J］.中国肺癌杂志，2011，14(8)：685-688.

MCFADDEN J W,CORL B A. Activation of AMP-activated protein kinase (AMPK) inhibits fatty acid synthesis in bovine mammary epithelial cells［J］. Biochem Biophys Res Commun, 2009, 390(3): 388-393.

HU H, WANG J, BU D, et al. In vitro culture and characterization of a mammary epithelial cell line from Chinese Holstein dairy cow［J］. PLoS ONE, 2009, 4(11): e7636.

胡小丽，张春林，王加启，等.不同剂量胰岛素、胰高血糖素和甲状腺素处理对奶牛乳腺上皮细胞FcRn mRNA表达的影响［J］.中国农业科学，2011，44(20)：4279-4283.

LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(Delta Delta C(T)) method［J］. Methods, 2001, 25(4): 402-408.

BIONAZ M, LOOR J J. Identification of reference genes for quantitative real-time PCR in the bovine mammary gland during the lactation cycle［J］. Physiol Genom, 2007, 29(3): 312-319.

胡菡，王加启，李发弟，等.高温诱导体外培养奶牛乳腺上皮细胞的应激响应［J］.农业生物技术学报，2011，19(2)：287-293.

BERNIER-DODIER P, GIRARD C L, TALBOT B G, et al. Effect of dry period management on mammary gland function and its endocrine regulation in dairy cows［J］. J Dairy Sci, 2011, 94(10): 4922-4936.

SHAO Y, WALL E H, MCFADDEN T B, et al. Lactogenic hormones stimulate expression of lipogenic genes but not glucose transporters in bovine mammary gland［J］. Domest Anim Endocrinol, 2013, 44 (2): 57-69.

LIU X, ROBINSON G W, HENNIGHAUSEN L. Activation of Stat5a and Stat5b by tyrosine phosphorylation is tightly linked to mammary gland differentiation［J］. Mol Endocrinol, 1996, 10(12): 1496-1506.

HENNIGHAUSEN L, ROBINSON G W. Interpretation of cytokine signaling through the transcription factors STAT5A and STAT5B［J］. Genes Dev, 2008, 22(6): 711-721.

SHEKAR P C, GOEL S, RANI S D, et al. kappa-casein-deficient mice fail to lactate［J］. Proc Natl Acad Sci U S A, 2006, 103(21): 8000-8005.

LE M N, KOHANSKI R A, WANG L H, et al. Dual mechanism of signal transducer and activator of transcription 5 activation by the insulin receptor［J］. Mol Endocrinol, 2002, 16(12): 2764-2779.

MENZIES K K, LEE H J, LEFEVRE C, et al. Insulin, a key regulator of hormone responsive milk protein synthesis during lactogenesis in murine mammary explants［J］. Funct Integr Genom, 2010, 10(1): 87-95.

HARRIS J, STANFORD P M, SUTHERLAND K, et al. Socs2 and elf5 mediate prolactininduced mammary gland development［J］. Mol Endocrinol, 2006, 20(5): 1177-1187.

LEMAY D G, NEVILLE M C, RUDOLPH M C, et al. Gene regulatory networks in lactation: identification of global principles using bioinformatics［J］. BMC Syst Biol, 2007, 1: 56.

ZHOU X, TAN M, STONE HAWTHORNE V, et al. Activation of the Akt/mammalian target of rapamycin/4EBP1 pathway by ErbB2 overexpression predicts tumor progression in breast cancers［J］. Clin Cancer Res, 2004, 10(20): 6779-6788.

KIMBALL S R, JEFFERSON L S. New functions for amino acids: effects on gene transcription and translation［J］. Am J Clin Nutr, 2006, 83(2): 500S-507S.

TOERIEN C A, CANT J P. Abundance and phosphorylation state of translation initiation factors in mammary glands of lactating and nonlactating dairy cows［J］. J Dairy Sci, 2007, 90(6): 2726-2734.

SALTIEL A R, PESSIN J E. Insulin signaling pathways in time and space［J］. Trends Cell Biol, 2002, 12(2): 65-71.

RAUGHT B, GINGRAS A C, SONENBERG N. The target of rapamycin (TOR) proteins［J］. Proc Natl Acad Sci U S A, 2001, 98(13): 7037-7044.

RIDER M H. The ubiquitin-associated domain of AMPK-related protein kinases allows LKB1induced phosphorylation and activation［J］. Biochem J, 2006, 394(Pt 3): e7-9.

黄德强，罗凌玉，王丽丽，等.AMPK在胰岛素信号转导通路中的作用［J］.中国细胞生物学学报，2011，33(11)：1220-1229.

刘微，邹俊杰，李晓曦，等.mTOR信号通路中上游调控蛋白Rheb在脂肪细胞分化中的作用［J］. 第二军医大学学报，2012，33(7)：697-702.

王红芳，杨维仁，刘建新，等.固醇调控元件结合蛋白及其对乳脂合成的调节作用［J］.动物营养学报，2010，22(5)：1165-1170.

SHIMOMURA I, BASHMAKOV Y, IKEMOTO S, et al. Insulin selectively increases SREBP-1c mRNA in the livers of rats with streptozotocin-induced diabetes［J］. Proc Natl Acad Sci U S A, 1999, 96(24): 13656-13661.

YANG Y A, HAN W F, MORIN P J, et al. Activation of fatty acid synthesis during neoplastic transformation: role of mitogen-activated protein kinase and phosphatidylinositol 3-kinase［J］. Exp Cell Res, 2002, 279(1): 80-90.

PETERSON D G, MATITASHVILI E A, BAUMAN D E. The inhibitory effect of trans-10, cis-12 CLA on lipid synthesis in bovine mammary epithelial cells involves reduced proteolytic activation of the transcription factor SREBP-1［J］. J Nutr, 2004, 134(10): 2523-2527.

林先滋，罗军，张犁苹，等. miR-200a对奶山羊乳腺上皮细胞乳脂合成相关基因mRNA表达的影响［J］.畜牧兽医学报，2012，43(7)：1028-1036.

[1]	ZHAO Shanjiang, SHUAI Zhiqiang, PANG Yunwei, HAO Haisheng, ZOU Huiying, DU Weihua, ZHAO Xueming, ZHU Huabin. Effect of Age at First Pregnancy on Milk Production and Reproductive Performance During the First 2 Lactations in Holstein Dairy Cattle [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(4): 732-742.
[2]	ZHAO Chang, ZHANG Jiang, BAI Yunlong, SUN Shuhan, SONG Yuxi, XIA Cheng. Analysis of Serum Differential Proteins in Cows with Inactive Ovaries Based on iTRAQ Technology [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(5): 972-982.
[3]	LI Xiao-jun, LI Xin, CHEN Xiao-li, ZHAO Yi-qiang, LU Yong-qiang, WANG Dong. The Transcription of Pld6 during Spermiogenesis and the Analysis of Its Protein Sequence in Dairy Cows [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(12): 2632-2640.
[4]	GUO Yan-sheng, TAO Jin-zhong. Selection of Milk Biomarkers of Pregnancy Recognition in Dairy Cows Based on LC-Q/TOF-MS Metabolomics [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(8): 1633-1641.
[5]	LIU Ying-chun, CHEN Yu-meng, HAN Zhe-xian, GAO Feng, ZHOU Huan-min. Induction and Identification of Neural Cell from Mammary Stem Cells in Holstein Cows and the Effect of Prolactin on Its Proliferative Activity [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(3): 542-548.
[6]	XU Chao, LI Feng, HAO Hai-sheng, LIU Yan, ZHAO Xue-ming, PANG Yun-wei, ZHU Hua-bin, DU Wei-hua. Sex-Related Genes Expression during Development of the Early Bovine Fetuses [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(10): 1892-1901.
[7]	ZHAO Yan-kun, SHAO Wei, WANG Li-wen, YU Xiong. Effect of Co-culturing with Umbilical Cord Mesenchymal Stem Cells on Milk Fat Synthesis and Expression of Key Genes in Bovine Mammary Gland Epithelial Cells [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(5): 846-853.
[8]	LIU Xiao-qian，LIU Ying, RONG Chao，WANG Kun，XIAO Hang，ZHANG Yuan-shu. Expression Localization of Angiotensin Converting Enzyme 2 (ACE 2) in Dairy Cow Mammary Gland and Its Anti-inflammation Effect [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(3): 552-560.
[9]	LI Xiao-jun，WANG Zhen-ling，CHEN Xiao-li，KOU Hong-xiang，LI Xiu-ling，WANG Yan，WANG Dong. Study Progress on the Rule of Body Temperature and Its Application in Reproduction of Dairy Cattle [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(12): 2331-2341.
[10]	CHENG Lei，WANG Ding-fa，LIU Xiao-hua，XIANG Min，LING Ming-hu，HU Xiu-zhong，XIA Yu. Study on ISG15 and OAS1 Transcriptions in Peripheral Blood of Dairy Cows during Early Pregnancy [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(1): 77-84.
[11]	WANG Xiao, XIE Xiao-li, WANG Sheng, QIAN Meng-ying, MA Pei-pei, ZHANG Yuan, SUN Dong-xiao, LIU Jian-feng, DING Xiang-dong, JIANG Li, WANG Ya-chun, ZHANG Yi, ZHANG Sheng-li, ZHANG Qin,YU Ying. Genome-wide Association Study for Mastitis Susceptibility and Resistance in Chinese Holsteins [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(12): 1907-1912.
[12]	ZHANG Mei-rong, CHEN Dan, LIAO Xiang-xiang, MAO Yong-jiang, LIU Kun, CHEN Liang, WANG Xing-long, YANG Zhang-ping, YANG Li-guo. Genetic Polymorphism of IL8 Gene and Its Association with Milk Traits and SCS in Holstein Imported from Australia [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(5): 690-696.

Effects of Insulin on mRNA Expression of Genes Related to Milk Protein and Fat Synthesis in Bovine Mammary Epithelial Cells Cultured in vitro

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 12

Recommended Articles

Metrics

Comments