甜菜碱对鹅胚原代肝细胞脂肪代谢和DNA甲基化的影响及调控机制  

Effects and Regulating Mechanism of Betaine on Lipid Metabolism and DNA Methylation in Primary Germinal Hepatocytes of Geese

作　　者：杨芷 杨雨 阳金金 王志跃 杨海明[2] YANG Zhi;YANG Yu;YANG Jingjing;WANG Zhiyue;YANG Haiming(Institutes of Agricultural Science and Technology Development,Yangzhou University,Yangzhou 225009,China;College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]扬州大学农业科技发展研究院,扬州225009 [2]扬州大学动物科学与技术学院,扬州225009

出　　处：《动物营养学报》2021年第5期2918-2926,共9页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：江苏现代农业(水禽)产业技术体系(JATS[2020]443);江苏省自然科学基金青年基金项目(BK20190897);扬州市自然科学基金青年基金项目(YZ2019080);江苏省研究生科研与实践创新计划(SJCX20_1382)。

摘　　要：本试验旨在研究甜菜碱对鹅胚原代肝细胞脂肪代谢和DNA甲基化的影响及调控机制。以鹅胚原代肝细胞为试验对象,试验1:研究不同浓度[0(对照)、2.5、5.0和10.0 mmol/L]甜菜碱培养液对鹅胚原代肝细胞DNA甲基化相关酶活性和泛酰巯基乙胺酶-1(VNN-1)启动子区域DNA甲基化的影响;试验2:研究不同处理[正常培养液(对照)、5.0μmol/L DNA甲基化酶抑制剂(AZA)、5.0 mmol/L甜菜碱、5.0μmol/L AZA+5.0 mmol/L甜菜碱]培养液对鹅胚原代肝细胞VNN-1和脂肪酸合成酶(FAS)基因表达的影响,探索VNN-1启动子区域甲基化调控基因表达的分子机制。结果表明:1)2.5、5.0和10.0 mmol/L甜菜碱组肝细胞的低密度脂蛋白(LDL)含量显著低于对照组(P<0.05)。2)S-腺苷甲硫氨酸/S-腺苷同型半胱氨酸(SAM/SAH)比值随着甜菜碱浓度的增加呈上升趋势,呈显著一次线性关系(P<0.05)。3)5.0和10.0 mmol/L甜菜碱组肝细胞VNN-1基因表达量显著低于对照组和2.5 mmol/L甜菜碱组(P<0.05);5.0和10.0 mmol/L甜菜碱组肝细胞VNN-1启动子区域DNA甲基化水平显著高于对照组(P<0.05)。4)AZA处理96 h时,AZA组和AZA+甜菜碱组肝细胞中VNN-1基因表达量显著高于对照组(P<0.05)。与对照组相比,处理48 h时,AZA处理显著提高了肝细胞中FAS基因表达量(P<0.05)。综上所述,鹅胚原代肝细胞培养液中添加甜菜碱能抑制肝细胞的脂质积累,提高细胞总体的DNA甲基化水平,且VNN-1启动子区域的甲基化负反馈调节其基因表达。AZA抑制DNA甲基化后发现VNN-1和FAS的高表达与DNA去甲基化相关。This experiment was conducted to study the effects and regulating mechanism of betaine on lipid metabolism and DNA methylation in primary germinal hepatocytes of geese.Geese primary germinal hepatocytes were used as the research material in this experiment.Experiment 1:this experiment was studied the effects of different concentrations[0(control),2.5,5.0 and 10.0 mmol/L]of betaine on DNA methylation related enzyme activity and DNA methylation in promoter region of VNN-1(pantetheinase)in geese primary germinal hepatocytes.Experiment 2:this experiment was investigated the effects of different treatments[normal culture medium(control),5.0μmol/L DNA methylation enzyme inhibitor(AZA),5.0 mmol/L betaine,5.0μmol/L AZA+5.0 mmol/L betaine]on the expression of VNN-1 and fatty acid synthase(FAS)genes in geese primary germinal hepatocytes,and explored the molecular mechanism of VNN-1 promoter region methylation regulates the molecular mechanism of gene expression.The results showed as follows:1)the content of low-density lipoprotein(LDL)in betaine culture medium containing 2.5,5.0 and 10.0 mmol/L was significantly lower than that in the control group(P<0.05).2)The S-adenosylmethionine/S-adenosylhomocysteine(SAM/SAH)ratio showed an increasing trend with increasing betaine concentration,with a significant primary linear relationship(P<0.05).3)The expression of VNN-1 gene in hepatocytes of 5.0 and 10.0 mmol/L betaine groups was significantly lower than that of control group and 2.5 mmol/L betaine group(P<0.05);DNA methylation in VNN-1 promoter region of hepatocytes of 5.0 and 10.0 mmol/L betaine groups was significantly higher than that of control group(P<0.05).4)The expression of VNN-1 gene in hepatocytes of AZA group and AZA+betaine group was significantly higher than that of the control group at 96 h of AZA treatment(P<0.05).AZA treatment significantly increased the expression of FAS gene in hepatocytes at 48 h of treatment compared with the control group(P<0.05).In conclusion,adding betaine to the primary germinal hepatoc

关 键 词：甜菜碱 鹅胚原代肝细胞 VNN-1 脂质代谢 DNA甲基化 

分 类 号：S816.7[农业科学—饲料科学]