Stimulation of glycerol kinase in grass carp preadipocytes by EPA
Stimulation of glycerol kinase in grass carp preadipocytes by EPA
复制标题
EPA对草鱼前脂肪细胞甘油激酶的刺激
DOI：
10.1007/s10695-016-0336-x
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发表时间：
2017
影响因子：
2.9
通讯作者：
Ji Hong
中科院分区：
文献类型：
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作者：
Lei Caixia;Tian Jingjing;Ji Hong
本研究旨在研究二十碳五烯酸(EPA)对草鱼前脂肪细胞甘油激酶(GyK)表达的影响，并探讨其作用机制。在此，我们克隆了草鱼GyK基因的部分序列，并分析了其组织分布。结果表明，GyK基因在肝脏中表达最多，其次是脂肪组织和肾脏。此外，用4 0 0μM油酸(18：1N-9，OA)建立肥大前体脂肪细胞模型。经100μ甲基二十碳五烯酸(20：5N-3，EPA)处理6、12、24小时后，GyK基因的表达和酶活性显著增强，同时，过氧化物体增殖激活受体γ、脂肪甘油三酯脂肪酶以及草鱼HSL基因的两种异构体被Sun等人首次鉴定，并将其定义为HSLA和HSLb。因此，HSLA和HSLB可能是合适的。EPA处理24小时后，甘油三酯含量显著增加，而竞争性拮抗剂HY-15859在12小时可减弱EPA诱导的GyK的升高。令人惊讶的是，血清剥夺引起的脂解和PPARγ基因表达的增强与GyK基因表达的增加是平行的，而GyK酶的活性则趋于稳定。其他脂肪酸，包括二十二碳六烯酸、α-亚麻酸、亚油酸和油酸也促进了GyK基因的表达。此外，我们还用一种不可逆的PPARγ拮抗剂GW9662来研究PPARγ在GyK诱导中的作用。数据显示，GW9662在12h取消了EPA对GyK的诱导，提示EPA上调了草鱼前脂肪细胞GyK的表达。ATgl和PPARγ参与了GyK的诱导。PPARγ可能是EPA诱导GyK表达的关键调节因子。
This study was conducted to assess the effect of eicosapentaenoic acid (EPA) on grass carp preadipocyte glycerol kinase (GyK) expression, as well as to explore the mechanism. Here, we cloned partial sequence of grass carp GyK gene and analyzed its tissue distribution. The result showed that GyK gene expressed most in the liver, followed by adipose tissue and the kidney. Besides, 400 μM oleic acid (18:1n-9, OA) was used to establish a hypertrophic preadipocyte model. GyK gene expression and enzyme activity were significantly enhanced after model cells were treated with 100 μM eicosapentaenoic acid (20:5n-3, EPA) for 6, 12, and 24 h. Meanwhile, peroxisome proliferative-activated receptor (PPAR)γ, adipose triglyceride lipase (ATGL), and the two isoforms of grass carp HSL gene were first identified by Sun et al (2016), and they defined the two isoforms as HSLa and HSLb. Therefore, maybe HSLa and HSLb are appropriate.. The content of triglyceride was dramatically increased by EPA treatment for 24 h. Further, a competitive ATGL antagonist, HY-15859, attenuated the increase in GyK induced by EPA at 12 h. Surprisingly, the enhanced lipolysis and PPARγ gene expression induced by serum deprivation were paralleled by an increase in GyK gene expression, whereas a stabilization in GyK enzyme activity. Other fatty acids, including docosahexaenoic acid, alpha-linolenic acid, linoleic acid, and OA also promoted GyK gene expression. Moreover, an irreversible PPARγ antagonist, GW9662, was used to investigate the role of PPARγ in GyK induction. Data showed that GW9662 abolished the induction of GyK by EPA at 12 h. Together, these data suggested that EPA elevated grass carp preadipocytes GyK expression. ATGL and PPARγ contributed to the induction of GyK. PPARγ may be a key regulator in response to GyK expression induced by EPA.