缬氨酸对猪原代肝细胞脂质代谢调节基因PPARα、FASN表达的影响  被引量:1

Effect of valine on expression of fatty acid metabolism-regulated genes PPARαand FASN in porcine primary hepatocytes

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作  者:谢宪兵[1] 黄仁祺 何玉珠 吴国云 陈星平 郑双艳[4] 万小娟 XIE Xianbing;HUANG Renqi;HE Yuzhu;WU Guoyun;CHEN Xingping;ZHENG Shuangyan;WAN Xiaojuan(Department of Laboratory Animal Science,Nanchang University,Nanchang 330031,China;The Second Clinical Medical College of Nanchang University,Nanchang 330031,China;College of Animal Science and Technology,Key Laboratory of Animal Nutrition in Jiangxi Province,Nanchang Key Laboratory of Animal Health and Safety,Jiangxi Agricultural University,Nanchang 330045,China;Sino-German Joint Research Institute,Nanchang University,Nanchang 330047,China;School of Basic Medical Sciences,Nanchang University,Nanchang 330006,China)

机构地区:[1]南昌大学实验动物科学中心,江西南昌330031 [2]南昌大学第二临床学院,江西南昌330031 [3]江西农业大学动物科学技术学院/江西省动物营养重点实验室/南昌市动物健康与安全重点实验室,江西南昌330045 [4]南昌大学中德联合研究院,江西南昌330047 [5]南昌大学基础医学院,江西南昌330006

出  处:《江西农业大学学报》2024年第2期427-437,共11页Acta Agriculturae Universitatis Jiangxiensis

基  金:国家自然科学基金项目(31960676);江西省自然科学基金项目(20202BABL215020,20212BAB205014,20232BAB215052)。

摘  要:【目的】研究旨在探究缬氨酸调节猪肝脏脂质代谢相关基因及其参与的调控途径,为揭示缬氨酸调控猪肝脏脂质代谢的分子作用机制提供试验数据和理论依据。【方法】以5日龄三元杂(杜洛克×长白×大白)仔猪原代肝细胞为试验对象,采用0,0.1,1,10 mmol/L缬氨酸(Val)分别处理原代肝细胞24 h及48 h,通过蛋白免疫印迹(Western-blot)技术检测不同浓度缬氨酸对脂质代谢重要调控基因PPARα及FASN的影响,同时将10 mmol/L缬氨酸处理24 h组与对照组的肝细胞样品进行RNA-Seq转录组测序,结合GO功能与KEGG通路富集分析,筛选出缬氨酸调控肝脏脂类代谢的潜在关键基因及信号通路,最后应用实时荧光定量(qPCR)技术验证差异表达基因。【结果】(1)与对照组相比,10 mmol/L缬氨酸处理原代肝细胞24 h和48 h均显著降低PPARα、FASN基因的蛋白表达(P<0.05);(2)与对照组相比,10 mmol/L缬氨酸处理原代肝细胞24 h组共筛选出418个差异表达基因,其中181个上调,237个下调。(3)GO功能分析发现,差异基因在分子功能方面主要参与蛋白质结合过程、催化活性、分子转导活性和转运蛋白活性。(4)KEGG分析发现,差异基因富集在代谢、疾病、生物系统、细胞信号调控、基因调控、环境调控等6个生物过程。其中脂质代谢是参与代谢生物过程的主要信号通路之一,富集到脂肪酸代谢的FASN、SCD、FADS2、FADS1、HSD17B12等5个差异基因主要参与胰岛素信号通路、AMPK信号通路、PPAR信号通路。【结论】10 mmol/L缬氨酸可显著下调猪原代肝细胞中PPARα及FASN的蛋白表达,调节脂质代谢关键基因的表达及信号通路活性,研究结果为进一步解析支链氨基酸调控脂代谢提供了理论基础。[Objective]This study aims to explore the genes related to the regulation of liver lipid metabolism by valine in pig and the regulatory pathways,providing experimental data and theoretical basis for revealing the molecular mechanism of of the regulation.[Method]The porcine primary hepatocytes derived from 5-day-old piglet(Duroc×Landrace×Large White)were treated with 0,0.1,1.0,and 10 mmol/L valine(Val)for 24 and 48 hours,respectively.The effects of different concentrations of valine on the important genes PPARαand FASN regulating lipid metabolism were detected through Western blot analysis.Potential key genes and signaling pathways involved in the regulation were screened.With GO function and KEGG pathway enrichment analysis,real-time fluorescence quantification(qPCR)technology was used to validate differentially expressed genes.[Result](1)The results showed that 10 mmol/L of valine significantly reduced the protein expression of PPARαand FASN(P<0.05)in porcine primary hepatocytes treated with valine for 24 and 48 hours,respectively.(2)The analysis of transcriptome sequencing data showed that compared with the control group,a total of 418 differentially expressed genes were screened in the valine group,of which 181 genes were upregulated and 237 genes were downregulated.(3)GO functional analysis revealed that differentially expressed genes mainly participated in Binding,Catalytic activity,Signal transducer activity,and Transporter activity in terms of molecular functions.(4)KEGG pathway enrichment analysis revealed that differentially expressed genes were enriched in six biological processes,including metabolism,human disease,organismal systems,cellular processes,genetic information processing,and environmental information processing.Lipid metabolism was one of the main signaling pathways involved in metabolism biological processes.The five differentially expressed genes enriched in fatty acid metabolism,including FASN,SCD,FADS2,FADS1,and HSD17B12,were mainly involved in the insulin signaling pathway,AMPK sign

关 键 词:缬氨酸  原代肝细胞 脂质代谢 转录组测序 

分 类 号:S852.2[农业科学—基础兽医学] S828[农业科学—兽医学]

 

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