限时摄食对AD模型小鼠脂肪LPL表达的调控及机制探讨  

EFFECTS OF TIME-LIMITED FEEDING ON LIPOPROTEIN LIPASE GENE EXPRESSION IN ADIPOSE TISSUE FROM MICE WITH ALZHEIMER’S DISEASE

作　　者：孟娜 李心蕙 张静竹[1] 潘鹏宇 胡爽 安丽[1] MENG Na;LI Xin-hui;ZHANG Jing-zhu;PAN Peng-yu;HU Shuang;AN Li(School of Public Health,China Medical University,Shenyang 110122;Liaoning Cancer Hospital and Institute,Shenyang 110042,China)

机构地区：[1]中国医科大学公共卫生学院,沈阳110122 [2]辽宁省肿瘤医院,沈阳110042

出　　处：《营养学报》2021年第4期362-369,共8页Acta Nutrimenta Sinica

基　　金：辽宁省教育厅科学研究一般项目(No.QN2019031)。

摘　　要：目的探讨限时摄食对AD模型小鼠脂肪LPL表达的调控作用及机制。方法将5月龄APPswe/PS1dE9双转基因AD模型小鼠及同窝出生的野生型小鼠分为AD模型和野生对照(自由摄食)、AD禁食和野生禁食共4组(摄食与禁食各24h交替进行),每组10只,连续饲养5月。再将小鼠前体脂肪(3T3-L1)细胞分为对照、βOHB、Aβ处理及βOHB+Aβ,其中βOHB组及βOHB+Aβ组细胞以1mmol/LβOHB预处理3h,再向Aβ处理组及βOHB+Aβ组细胞加入0.5或2μmol/LAβ,继续培养12h。最后,以siRNA沉默组蛋白去乙酰化酶2/3(siRNA-HDAC2/siRNA-HDAC3),同时设立对照(scrambled siRNA)组。采用qRT-PCR和Western blotting法检测脂肪组织和3T3-L1细胞LPL、HDAC2/3mRNA及蛋白表达水平,Western blotting法检测脂肪组织及3T3-L1细胞组蛋白乙酰化(Ace-H3K9/H4K12)水平。结果限时摄食可抑制AD模型小鼠脂肪组织LPL表达水平的下调和Ace-H3K9/H4K12水平的降低,以及HDAC2/3表达水平的升高。0.5μmol/L Aβ处理的3T3-L1细胞LPL表达水平升高,而2μmol/L Aβ处理的3T3-L1细胞LPL表达水平则降低;βOHB可抑制0.5或2μmol/L Aβ处理的3T3-L1细胞LPL表达水平的变化,以及Ace-H3K9/H4K12水平的降低和HDAC2/3表达水平的升高。沉默细胞HDAC2或HDAC3可分别下调或上调LPL表达。结论限时摄食可调控AD模型小鼠脂肪组织LPL的表达,βOHB对HDAC2、HDAC3的抑制可能参与上述调控过程。Objective To investigate the effects of time-limited feeding on LPL expression in adipose tissue from AD model mice.Methods APPswe/PS1 dE9 double-transgenic mice(5-month-old)were assigned into AD and AD+ADF group,and their wild-type littermates into WT and WT+ADF group(5 males and 5 females in each group).Mice in WT+ADF and AD+ADF groups were treated with time-limited feeding(alternated feeding and fasting for 24 hours each).Mice in WT and AD groups were fed ad libitum.After 5 months of experiment,the adipose tissue was collected for analyses.Precursor fat(3 T3-L1)cells were cultured and then divided into four groups:control,βOHB,AβandβOHB+Aβ.Cells inβOHB andβOHB+Aβgroups were pretreated with a final concentration of 1 mmol/LβOHB for 3 hours.Subsequently,cells in AβandβOHB+Aβgroups were treated with a final concentration of 0.5 or 2μmol/L Aβ.Twelve hours later,cells were collected for assay.Finally,the endogenous HDAC2 and HDAC3 mRNA levels in cells were interfered by HDAC2 and HDAC3 siRNA duplex respectively and scrambled siRNA was used as control.The relative expression levels of LPL,HDAC2 and HDAC3 mRNA in the adipose tissue and 3 T3-L1 cells were tested using quantitative real-time polymerase chain reaction.The relative expression levels of LPL,HDAC2,HDAC3 protein and Ace-H3 K9/H4 K12 levels in the adipose tissue and 3 T3-L1 cells were detected by Western blotting.Results Time-limited feeding exhibited inhibitory effects on the down-regulation of LPL expression,the decrease of Ace-H3 K9/H4 K12 levels,and the up-regulation of HDAC2/3 expression in AD mice.The expression level of LPL was increased in 3 T3-L1 cells treated with a final concentration of 0.5μmol/L Aβ,while increased in cells treated with 2μmol/L Aβ.βOHB inhibited the alterations of LPL expression levels,the decrease of Ace-H3 K9/H4 K12 levels and the up-regulation of HDAC2/3 expression levels in 3 T3-L1 cells with 0.5μmol/L or 2μmol/L Aβtreatment.The expression levels of LPL were down-regulated and up-regulated,respectively,in

关 键 词：阿尔茨海默病 Β-淀粉样蛋白 限时摄食 脂蛋白脂酶 

分 类 号：R152.2[医药卫生—营养与食品卫生学]