代谢性疾病易感猪皮下脂肪功能障碍分子病理机制解析  

Molecular Pathological Mechanisms of Subcutaneous Fat Dysfunction in Metabolic Disease Susceptible Pigs

作　　者：徐塽 杜娟 张凯艺 苗佳坤 杨宇 王彦芳[1] 杨述林[1] XU Shuang;DU Juan;ZHANG Kaiyi;MIAO Jiakun;YANG Yu;WANG Yanfang;YANG Shulin(State Key Laboratory of Animal Biotech Breeding,Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100193,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所畜禽生物育种全国重点实验室,北京100193

出　　处：《畜牧兽医学报》2024年第11期4938-4949,共12页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(32070535);国家重点研发计划(2021YFA0805903);中国农业科学院科技创新工程(ASTIP-IAS05)。

摘　　要：旨在了解代谢性疾病易感猪皮下脂肪功能障碍的分子病理机制,探究皮下脂肪组织能量代谢紊乱与表观调控的相关性。本研究选用体重相近、健康状况良好的6月龄野生型雄性巴马猪个体及转基因制备的代谢性疾病易感猪,经高脂高糖饮食(high-fat high-sugar diet,HFHSD)诱导3个月或12个月后,将其分为4组,分别为饮食诱导3个月的野生型组(WT-3,n=5)、饮食诱导12个月的野生型组(WT-12,n=5)、饮食诱导3个月的转基因组(TG-3,n=8)和饮食诱导12个月的转基因组(TG-12,n=4)。首先对试验动物进行血清学评价,检测甘油三酯、游离脂肪酸、瘦素和脂联素浓度;然后对试验动物进行组织学病理评价,并且通过转录组测序获得代谢紊乱的分子特征及富集的信号通路;随后通过qPCR、蛋白免疫印迹(Western blot,WB)、酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)对关键基因、代谢物及表观修饰进行检测。血清生化测定结果及病理组织学评价表明,转基因及饮食诱导均能引起脂肪组织功能障碍,转基因联合饮食诱导组的病理损伤更严重;转录组测序结果表明,脂肪组织功能障碍典型的特征是线粒体氧化磷酸化功能受损,脂肪组织糖脂代谢和蛋白质合成等功能下降;qPCR结果显示,TG-12组中线粒体编码的基因表达显著下调;WB结果显示,TG-12组中调节糖脂代谢的关键基因ACLY、ACSS2和FASN显著下调;ELISA结果显示,关键的中间代谢物乙酰辅酶A含量降低;且经WB验证,在基因表达中具有广泛调控作用的组蛋白乙酰化水平降低,可能是脂肪功能障碍的主要原因。利用代谢性疾病易感猪揭示了皮下脂肪线粒体功能降低通过下调乙酰辅酶A含量及组蛋白乙酰化水平,经表观修饰发挥广泛的基因表达抑制作用,为人类肥胖相关皮下功能障碍疾病治疗提供了模型和参考数据。The aim of this study was to understand the molecular pathological mechanisms of subcutaneous adipose dysfunction in pigs susceptible to metabolic diseases,and to investigate the correlation between disturbances of energy metabolism in subcutaneous adipose tissue and epigenetic regulation.The study selected wild-type male Bama pigs with similar weight and good health at 6 months old,as well as transgenic pigs susceptible to metabolic diseases,and divided them into 4 groups after being induced by a high-fat high-sugar diet(HFHSD)for 3 or 12 months.The wild-type groups were named WT-3(n=5)and WT-12(n=5),while the transgenic groups were named TG-3(n=8)and TG-12(n=4).Firstly,the animals were evaluated by serum biochemistry,containing the concentrations of triglycerides,free fatty acids,leptin,and adiponectin.Then,the animals were subjected to histopathological evaluation,and molecular features and enriched signaling pathways of metabolic disorders were obtained through transcriptome sequencing.Subsequently,key genes,metabolites,and epigenetic modifications were detected using qPCR,Western blot,and ELISA.The results of serum biochemical assay and histopathological evaluation showed that both transgenic and diet induction could cause adipose tissue dysfunction,and the pathological injury of transgenic combined diet induction group was more serious.Transcriptome sequencing results showed that adipose tissue dysfunction was characterized by impaired mitochondrial oxidative phosphorylation and decreased glucose and lipid metabolism and protein synthesis in adipose tissue.qPCR results showed that the expression of mitochondria-encoded genes was significantly down-regulated in TG-12 group.WB results showed that the key genes regulating glucose and lipid metabolism,ACLY,ACSS2 and FASN,were significantly down-regulated in TG-12 group.ELISA results showed that the content of acetyl-coA,the key intermediate metabolite,was decreased.Moreover,WB verified that the reduction of histone acetylation,which has a wide regulatory role i

关 键 词：转基因猪 皮下脂肪组织 转录组 线粒体 组蛋白乙酰化 

分 类 号：S852.21[农业科学—基础兽医学]