DLD和NDUFA9及其巴豆酰化促进仔猪原代皮下白脂棕色化  

DLD,NDUFA9 and Their Crotonylation Promote Browning of Primary Subcutaneous White Adipocytes in Piglets(Sus scrofa)

作　　者：刘月霞[1] 刘尊海 梁君桐 陈齐 陈欣豪 左云非 孙超[1] LIU Yue-Xia;LIU Zun-Hai;LIANG Jun-Tong;CHEN Qi;CHEN Xin-Hao;ZUO Yun-Fei;SUN Chao(College of Animal Science and Technology,Northwest A&F University,Yangling,712100,China)

机构地区：[1]西北农林科技大学动物科技学院,杨凌712100

出　　处：《农业生物技术学报》2024年第4期820-832,共13页Journal of Agricultural Biotechnology

基　　金：国家十四五重点研发计划(2021YFF1000602);陕西省重点研发项目(2021NY-020);陕西省创新能力支撑计划(2023-CX-TD-57);科技援青合作专项(2022-QY-219)。

摘　　要：二氢硫辛酸脱氢酶(dihydrolipoyl dehydrogenase,Dld)和烟酰胺腺嘌呤二核苷酸(nicotinamide adenine dinucleotide,NADH)脱氢酶[泛醌] 1 α亚复合体亚基9(NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 9,Ndufa9)基因均与能量代谢和线粒体功能相关,提示Dld和Ndufa9与白脂棕色化可能存在内在联系。本研究以仔猪原代皮下白色脂肪细胞为研究对象,诱导其分化为成熟脂肪细胞。分别使用Dld与Ndufa9干扰片段和过表达载体处理成熟脂肪细胞后,利用实时定量PCR(real-time quantitative PCR,q PCR)和Western blot检测脂质代谢相关基因表达水平,结果显示,过表达Dld与Ndufa9后,脂合成相关基因脂肪酸结合蛋白4(fatty acid binding protein 4,Fabp4)和脂肪酸合成酶(fatty acid synthase,Fasn)显著降低(P<0.05),脂解相关基因脂肪甘油三酯脂酶(adipose triglyceride lipase,Atgl)和激素敏感脂肪酶(hormone sensitivelipase,Hsl)、白脂棕色化相关基因过氧化物酶体增殖激活受体γ共激活因子1α(peroxisome proliferator-activated receptor gamma coactivator 1 alpha,Pgc-1α)和含PR结构域的锌指蛋白16(PR domain containing 16,Prdm16)、产热相关基因解耦蛋白2(uncoupling protein 2,Ucp2)、骨形态发生蛋白4(bone morphogenetic proteins 4,Bmp4)、Bmp7和诱导细胞死亡的DNA片段化因子α样效应因子A(cell death-inducing DNA fragmentation factor α-like effector A,Cidea)均显著上调(P<0.05),表明Dld与Ndufa9均促进仔猪皮下白色脂肪细胞发生棕色化。使用伏立诺他(vorinostat,SAHA)和巴豆酸钠(NaCr)处理成熟脂肪细胞。发现DLD和NDUFA9巴豆酰化同样促进仔猪皮下白色脂肪细胞棕色化。通过免疫共沉淀(co-immunoprecipitation,Co-IP)检测发现DLD和NDUFA9结合。以上结果提示,DLD和NDUFA9及其巴豆酰化修饰促进仔猪原代皮下白脂棕色化,也表明Dld和Ndufa9是调控猪脂质代谢的关键因子。本研究为进一步探究猪肉品质改良提供理论依据。Dihydrolipoyl dehydrogenase(Dld) and nicotinamide adenine dinucleotide(NADH) dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 9(Ndufa9) gene were found to be related to energy metabolism and mitochondrial function.It was suggested that Dld and Ndufa9 may be intrinsically linked to white adipocytes browning.This study aimed to investigate the effect of Dld,Ndufa9 and their crotonylation modifications on the browning of primary subcutaneous white adipocytes in piglets(Sus scrofa).To investigate this intrinsic linkage,this study was conducted with primary subcutaneous white adipocytes from piglets,and induced their differentiation into mature adipocytes.Interference and overexpression treatment of Dld and Ndufa9,detection of expression of lipid metabolism genes using real-time quantitative PCR(qPCR) and Western blot.The results revealed that lipogenesis-related genes fatty acid binding protein 4(Fabp4) and fatty acid synthase(Fasn) were significantly lower(P<0.05) and lipolysis-related genes adipose triglyceride lipase(Atgl) and hormone sensitivelipase(Hsl),browning of white fat-related genes peroxisome proliferatoractivated receptor gamma coactivator 1 alpha(Pgc-1α) and PR domain containing 16(Prdm16) and thermogenesis-related genes uncoupling protein 2(Ucp2),bone morphogenetic proteins 4(Bmp4),bone morphogenetic proteins 7(Bmp7) and cell death-inducing DNA fragmentation factorα-like effector A(Cidea)were significantly up-regulated(P<0.05) after overexpression of Dld and Ndufa9 compared with the control group,which indicated that Dld and Ndufa9 promoted browning of subcutaneous white adipocytes in piglets.Next,mature adipocytes were treated with vorinostat(SAHA) and sodium crotonate(NaCr) to give the cells a crotonylation-modified environment.It was found that crotonylation modification of Dld and Ndufa9 similarly promoted browning of subcutaneous white adipocytes in piglets.And through co-immunoprecipitation(CoIP),it was further discovered that DLD combined with NDUFA9.These results suggested that DLD,NDUFA

关 键 词：仔猪原代皮下白色脂肪细胞 白脂棕色化 二氢硫辛酸脱氢酶(DLD) NADH脱氢酶[泛醌]1α亚复合体亚基9(NDUFA9) 巴豆酰化修饰 

分 类 号：S813.3[农业科学—畜牧学]