摘要
过多的脂肪和胆固醇随着肉蛋奶被人体摄入是导致人类高血脂等各种疾病诱发的原因之一,而探索脂代谢通路相关基因的表达变化及其调控机制已经成为分子生物学技术在兽医学领域中的研究热点。与高血脂有关的ApoE、ApoC1和Tomm40等基因研究较多,而与这些基因共同位于小鼠七号染色体上且遗传距离较近的PVRL-2及其编码蛋白Nectin-2则少见报道。为了解PVRL-2在脂代谢中的作用,本研究利用成功构建的PVRL-2慢病毒载体包装病毒在3T3-L1前脂细胞中进行了过表达分析,并通过生物信息学方法对鼠Nectin-2蛋白的结构及其在血脂调控方面的潜在功能进行了预测。结果显示,与对照相比,病毒侵染的3T3-L1细胞中PVRL-2的表达量升高近100倍,而PPARα的表达量则升高了4.5倍;蛋白结构预测显示Necitn-2为含有信号肽的跨膜蛋白。PPARα表达量上升会引起脂肪酸代谢相关基因的活化进而能够改善机体的脂代谢,而信号肽以及跨膜结构的存在表明Nectin-2蛋白有可能是通过与膜上的蛋白受体相结合进而启动下游脂代谢的信号通路。这提示或可将Nectin-2作为新的预防和治疗血脂异常等心血管疾病的药物靶点。
Excess fat and cholesterol in food such as meat,eggs or milk could lead to hyperlipoidemia in human.Currently,to explore genes expression and their mechanisms associated with lipid metabolism has been a major focus in veterinary science.Growing bodies of evidence indicated that molecular functions of fatty acid metabolism related genes such as ApoE,ApoC1 and Tomm40 were very well characterized;however,function of their chromosomal neighbor such as PVRL-2 gene in the fatty acid metabolism remains unclear.Present study was aim to investigate potential role of mouse PVRL-2 gene in regulation of fatty acid related gene expression using preadipogenic 3T3-L1 cells.The cells were infected by Lentiviral particles which was produced by lentiviral plasmid containing Pvrl2 gene,and RNA were extracted 48h post viral infection.Quantitative real-time PCR analysis confirmed that PVRL-2 overexpressed more than 100 folds upon PVRL-2 virus transformation compared to the control.Notably,the expression of PPARα gene which is a key player in the fatty acid oxidation was strongly induced（4.5 fold increase） post PVRL-2 viral infection,but not other genes that related to the fatty acid metabolism such as CPT1A,FASN,COX7Α,PGC1B,ASADM showed similar changes.Furthermore,bioinformatics analyses revealed that Nectin-2,coded by PVRL-2,should be a transmembrane protein with a signal peptide.In conclusion,the present study demonstrated that overexpression of PVRL-2 induce the expression of PPARα,which highlight the potential roles of PVRL-2 gene in fatty acid metabolism.Future studies are needed to determine detailed molecular function of PVRL-2 gene in fatty acid metabolism.
出处
《畜牧与兽医》
北大核心
2013年第8期8-13,共6页
Animal Husbandry & Veterinary Medicine
基金
中国科学院百人计划启动基金(Y151541104)
