摘要
溶血磷脂酸酰基转移酶(Sn-1-acylglycerol-3-phosphate O-acyltransferase 6,AGPAT6)是脂类物质合成代谢相关酶,但其在奶牛乳脂合成中调节作用尚不明确。研究采用荧光定量PCR、Western blot等方法检测奶牛泌乳循环过程中乳腺组织中AGPAT6表达变化、奶牛乳腺上皮细胞中AGPAT6对乳脂合成影响以及相关信号通路分子表达变化。结果表明,AGPAT6 mRNA和蛋白质表达在泌乳期奶牛乳腺组织中极显著高于青春期和干奶期(P<0.01)。在具有乳脂合成能力的奶牛乳腺上皮细胞中,AGPAT6基因过表达极显著提高细胞中TAG含量,AGPAT6基因干扰获得相反结果。AGPAT6过表达可激活PI3K-AKT-mTOR信号通路,添加PI3K抑制剂会抑制AGPAT6诱导PI3K-AKT-mTOR信号通路激活及细胞中TAG合成(P<0.01)。以上结果显示AGPAT6表达与泌乳奶牛乳脂合成呈正相关,AGPAT6通过激活PI3K-AKT-mTOR信号通路促进奶牛乳腺上皮细胞中乳脂合成。
Sn-1-acylglycerol-3-phosphate O-acyltransferase 6(AGPAT6)is a key enzyme related to TAG synthesis;however,the role of AGPAT6 in milk fat synthesis in dairy cow is not well understood.In this study,the expression of AGPAT6 in mammary tissues of dairy cows during lactation cycle was detected by fluorescence quantitative PCR and Western blot,and then,the effects of AGPAT6 on signaling pathway activation and milk fat synthesis were also detected.The results showed that the mRNA and protein level of AGPAT6 was significantly higher in mammary tissues of lactating cows than those in puberty and dry cows(P<0.01).In cultured mammary epithelial cells of dairy cows,AGPAT6 overexpression extremely significantly increased the intracellular TAG content,while AGPAT6 knockdown had the opposite effect(P<0.01).Western blot analysis showed that AGPAT6 overexpression activated PI3K-AKT-mTOR signaling pathway,and this activation and AGPAT6-induced TAG synthesis were inhibited when the cells were treated with PI3K inhibitor.Taken together,these data suggested that AGPAT6 expression was positively correlated with milk fat synthesis in mammary glands of lactating cows.AGPAT6 regulated milk fat synthesis via PI3K-AKT-mTOR signaling pathway.
作者
林叶
刘灿
江怡
宫晓庆
刘川萍
杨洋
侯晓明
LIN Ye;LIU Can;JIANG Yi;GONG Xiaoqing;LIU Chuanping;YANG Yang;HOU Xiaoming(Heilongjiang Key Laboratory of Animal Cell and Genetic Engineering,Northeast Agricultural University,Harbin 150030,China)
出处
《东北农业大学学报》
CAS
CSCD
北大核心
2022年第4期41-48,共8页
Journal of Northeast Agricultural University
基金
国家自然科学基金面上项目(31771453,31671285)
东北农业大学“学术骨干”基金资助项目(18XG25)。
