摘要
旨在克隆获得山羊PDK4基因序列,明确其组织细胞表达特性,并明晰其对山羊肌内脂肪细胞脂代谢的作用。构建山羊PDK4过表达和干扰细胞模型,利用RT-PCR、实时荧光定量PCR等研究PDK4对山羊肌内脂肪细胞脂代谢的影响。结果克隆获得山羊PDK4基因序列,长度为1808 bp,定位于线粒体和细胞质。明确了山羊PDK4组织及细胞时序表达模式,发现PDK4高表达于山羊肺脏、臂三头肌和肝脏组织(P<0.01);在诱导分化5 d的山羊肌内脂肪细胞中表达水平极显著高于诱导分化之前的表达水平(P<0.01)。干扰和过表达山羊PDK4分别显著降低和增加了脂质积聚,且干扰PDK4后脂代谢相关基因FABP3、CD36、ACACA、AGPAT6和ADRP表达水平均显著(P<0.05)或极显著(P<0.01)降低,而过表达PDK4后表达水平极显著升高(P<0.01)。PDK4在山羊各组织及肌内前体脂肪细胞分化各个阶段均存在表达。过表达山羊PDK4促进了脂肪细胞脂滴累积和脂质沉积相关基因表达上调,干扰后呈相反的趋势和表达。
This study aims to clone the goat PDK4 gene sequence,to clarify its tissue and temporal expression characteristics,and to clarify its effect on the lipid metabolism of goat intramuscular adipocytes.Overexpression and interference cell model of goat PDK4 was constructed,and RT-PCR and real-time fluorescent quantitative PCR were used to determine and study the effect of PDK4 on the lipid metabolism in goat intramuscular adipocytes.As results,the cloned goat PDK4 sequence was 1808 bp,and it was located in mitochondria and cytoplasm.The expression pattern of goat PDK4 tissue and cell temporal expression was clarified,showing that PDK4 was highly expressed in the goat’s lung,triceps brachii muscle and liver tissue(P<0.01).The expression level in goat intramuscular adipocytes at 5 d of induction was significantly higher than that before differentiation(P<0.01).Interference and overexpression of goat PDK4 significantly reduced and increased lipid accumulation,respectively.The expression of lipid metabolism-related genes FABP3,CD36,ACACA,AGPAT6 and ADRP significantly(P<0.05)or extremely significantly(P<0.01)decreased with si-PDK4,while those genes extremely significantly increased(P<0.01)after overexpression of PDK4.In conclusion,PDK4 is expressed in goat tissues and in various stages of intramuscular preadipocyte differentiation.Overexpression of goat PDK4 promots the up-regulation of lipid droplet accumulation and lipid deposition-related gene expression in adipocytes,and shows the opposite trend and expression after interference.
作者
张浩
张亚楠
李鑫
王佳美
王永
朱江江
熊燕
林亚秋
ZHANG Hao;ZHANG Ya-nan;LI Xin;WANG Jia-mei;WANG Yong;ZHU Jiang-jiang;XIONG Yan;LIN Ya-qiu(Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Protection and Utilization of Ministry of Education/Sichuan Province,Southwest Minzu University,Chengdu 610041;College of Animal&Veterinary Sciences,Southwest Minzu University,Chengdu 610041)
出处
《生物技术通报》
CAS
CSCD
北大核心
2021年第12期151-159,共9页
Biotechnology Bulletin
基金
国家自然科学基金项目(31902154)
四川省杰出青年科技人才计划项目(2020JDJQ0010)。
关键词
丙酮酸脱氢酶激酶4
山羊
表达特性
脂代谢
pyruvate dehydrogenase kinase 4
goat
expression characteristics
lipid metabolism
