李君;梁文双;赫秋亚;杨芳慧;权凯;王笑笑;邓红雨;
LI Jun;LIANG Wenshuang;HE Qiuya;YANG Fanghui;QUAN Kai;WANG Xiaoxiao;DENG Hongyu;Henan International Joint Laboratory of Ruminant Nutrition and Feed Resources Development, College of Animal Science and Technology, Henan University of Animal Husbandry and Economy;College of Animal Science and Technology, Northwest Agriculture and Forestry University;

• 1:河南牧业经济学院动物科技学院河南省反刍动物营养与饲料资源开发国际联合实验室
• 2:西北农林科技大学动物科技学院

摘要(Abstract)：

为研究SIRT1/FoxO1通路在奶山羊乳腺脂质合成中的作用,本实验采用0(对照)、50、100、150μmol/L的SIRT1激动剂(RES)处理奶山羊乳腺上皮细胞,qRT-PCR检测脂质代谢相关基因的表达,检测细胞中甘油三酯含量,油红O染色法观察脂滴的积累情况。结果表明:100μmol/L RES处理组的SIRT1和FoxO1基因的相对mRNA表达量高于对照组(P<0.05),乳脂关键调控因子SREBP1和PPARγ表达量下降(P<0.05);SIRT1/FoxO1通路激活后,脂肪酸合酶FASN(P<0.01)、脂肪酸去饱和酶SCD1(P<0.01)和脂肪酸转运酶CD36(P<0.05)表达量均下降;而脂解相关基因HSL和ATGL的表达上调(P<0.05)。SIRT1/FoxO1激活后,细胞中甘油三酯含量显著降低(P<0.05),脂滴积累减少。综上可知,SIRT1/FoxO1通路负调控奶山羊乳腺上皮细胞脂质合成,为改善羊奶营养价值和风味提供基础资料。
To study the effects of SIRT1/FoxO1 pathway on lipid metabolism in dairy goat mammary gland. The goat mammary epithelial cells were treated with 0(control), 50, 100, 150 μmol/L of SIRT1 agonist(RES), the expression of genes related to lipid metabolism was detected by qRT-PCR, the content of triglycerides was detected by enzymatic colorimetric, and the accumulation of lipid droplets in cells was observed by oil red O staining. The results showed that the relative expression of SIRT1 and FoxO1 genes in 100 μmol/L RES group were higher than that of the control group. After activation of SIRT1/FoxO1 pathway, the expression of SREBP1, PPARγ, FASN, SCD1 and CD36 were lower than those of the control group(P<0.01,P<0.05). Meanwhile, the expression of HSL and ATGL were significantly up-regulated(P<0.05). After activation of SIRT1/FoxO1 pathway, the content of triglycerides and lipid droplets were significantly decreased(P<0.05). In summary, the findings indicated that SIRT1/FoxO1 have negative regulatory effect on lipid synthesis in dairy goat mammary epithelial cells, providing basic data for improving the nutritional value and flavor of goat milk.

关键词(KeyWords)： SIRT1;乳腺上皮细胞;脂质合成;甘油三酯;脂滴积累
SIRT1;Mammary gland epithelial cells;Lipid synthesis;Triglycerides;Lipid droplet accumulation

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(31601915);; 河南省高等学校重点科研项目(17A230012);; 河南牧业经济学院博士科研启动项目

作者(Author): 李君;梁文双;赫秋亚;杨芳慧;权凯;王笑笑;邓红雨;
LI Jun;LIANG Wenshuang;HE Qiuya;YANG Fanghui;QUAN Kai;WANG Xiaoxiao;DENG Hongyu;Henan International Joint Laboratory of Ruminant Nutrition and Feed Resources Development, College of Animal Science and Technology, Henan University of Animal Husbandry and Economy;College of Animal Science and Technology, Northwest Agriculture and Forestry University;

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参考文献(References)：

• [1]Cao Y,Jiang X,Ma H,et al.SIRT1 and insulin resistance[J].JDiabetes Complications,2016,30(1):178-183.
• [2]Motta M C,Divecha N,Lemieux M,et al.Mammalian SIRT1represses forhead transcription factors[J].Cell,2004,116(4):551-563.
• [3]Picard F,Kurtev M,Chung N,et al.Sirt1 promotes fat mobilization in white adipocytes by repressing PPAR-gamma[J].Nature,2004,429(6993):771-776.
• [4]Zhao H,Chen S,Gao K,et al.Resveratrol protects against spinal cord injury by activating autophagy and inhibiting apoptosis mediated by the SIRT1/AMPK signaling pathway[J].Neuroscience,2017,348:241-251.
• [5]Rodgers J T,Lerin C,Haas W,et al.Nutrient control of glucose homeostasis through a complex of PGC-1alpha and SIRT1[J].Nature,2005,434(7029):113-118.
• [6]Xu H F,Luo J,Zhao W S,et al.Overexpression of SREBP1(sterol regulatory element binding protein 1)promotes de novo fatty acid synthesis and triacylglycerol accumulation in goat mammary epithelial cells[J].J Dairy Sci,2016,99(1):783-795.
• [7]Shi H,Zhao W,Zhang C,et al.Transcriptome-wide analysis reveals the role of PPARγcontrolling the lipid metabolism in goat mammary epithelial cells[J].PPAR Res,2016,2016(9):1-11.
• [8]Shen Z,Liang X,Rogers C Q,et al.Involvement of adiponectinSIRT1-AMPK signaling in the protective action of rosiglitazone against alcoholic fatty liver in mice[J].Am J Physiol,2010,298(1):364-374.
• [9]Yasar C,Oguzhan O,Ebubekir S,et al.SIRT1 as a potential therapeutic target for treatment of nonalcoholic fatty liver disease[J].Med Sci Monit,2011,17(5):5-9.
• [10]Anderson S M,Rudolph M C,Mcmanaman J L,et al.Key stages in mammary gland development.Secretory activation in the mammary gland:it's not just about milk protein synthesis![J].Breast Cancer Res Bcr,2007,9(1):204.
• [11]Bionaz M,Loor J J.Gene networks driving bovine milk fat synthesis during the lactation cycle[J].Bmc Genomics,2008,9(1):366-387.
• [12]Chakrabarti P,English T,Karki S,et al.SIRT1 controls lipolysis in adipocytes via FOXO1-mediated expression of ATGL[J].J Lipid Res,2011,52(9):1693-1701.
• [13]Yao D,Luo J,He Q,et al.SCD1 alters long-chain fatty acid(LCFA)composition and its expression is directly regulated by SREBP-1 and PPARγ1 in dairy goat mammary cells[J].J Cell Physiol,2016,232(3):635-649.
• [14]Huang C H,Shiu S M,Wu M T,et al.Monacolin K affects lipid metabolism through SIRT1/AMPK pathway in HepG2 cells[J].Arch Pharm Res,2013,36(12):1541-1551.