中华老年医学杂志
中華老年醫學雜誌
중화노년의학잡지
Chinese Journal of Geriatrics
2013年
7期
773-777
,共5页
李国平%唐蔚青%隋靖喆%黎健%陈保生
李國平%唐蔚青%隋靖喆%黎健%陳保生
리국평%당위청%수정철%려건%진보생
肝细胞%受体,胰岛素%脂蛋白类,VLDL%载脂蛋白B类%三酰甘油类
肝細胞%受體,胰島素%脂蛋白類,VLDL%載脂蛋白B類%三酰甘油類
간세포%수체,이도소%지단백류,VLDL%재지단백B류%삼선감유류
Hepatocytes%Receptor,insulin%Lipoprotein,VLDL%Apolipoproteins B%Triglycerides
目的 分析肝脏组织特异性的胰岛素信号急性缺失对载脂蛋白B和三酰甘油分泌的影响. 方法 应用Cre-LoxP系统原理,在重组酶基因的上游插入肝组织特异性白蛋白基因启动子,以此构建组织特异性表达的腺病毒.扩增纯化后的腺病毒经胰岛素受体Floxed小鼠尾静脉注射,分别在注射2d、4d和6d后收集小鼠血浆,测定新生载脂蛋白B和极低密度脂蛋白三酰甘油分泌速率,抽提肝脏脂质并用酶法检测脂质含量,并用免疫印迹法分析胰岛素受体和脂代谢相关蛋白在肝脏内的表达. 结果 小鼠在注射含重组酶的腺病毒2d、4d和6d后,胰岛素受体表达依次降低30.50％ (P＜0.05)、60.12％ (P＜0.01)和99.54％ (P＜0.001),极低密度脂蛋白三酰甘油的分泌速率分别降低20.43％ (P＜0.05)、33.63％(P＜0.05)和44.21％(P＜0.01);下调了固醇调控结合蛋白1和脂肪酸合成酶以及极低密度脂蛋白形成相关蛋白的表达,但不影响新生载脂蛋白B100的分泌和肝脏脂质含量；而在含重组酶的腺病毒注射6d后新生载脂蛋白B48升高约35.07％ (P＜0.05).结论 肝脏胰岛素信号急性缺失降低极低密度脂蛋白中三酰甘油的分泌速率并呈现时间依赖性,揭示小鼠肝脏分泌较小的极低密度脂蛋白颗粒,这种变化可能和肝脏内脂肪酸的合成速度下降有关.
目的 分析肝髒組織特異性的胰島素信號急性缺失對載脂蛋白B和三酰甘油分泌的影響. 方法 應用Cre-LoxP繫統原理,在重組酶基因的上遊插入肝組織特異性白蛋白基因啟動子,以此構建組織特異性錶達的腺病毒.擴增純化後的腺病毒經胰島素受體Floxed小鼠尾靜脈註射,分彆在註射2d、4d和6d後收集小鼠血漿,測定新生載脂蛋白B和極低密度脂蛋白三酰甘油分泌速率,抽提肝髒脂質併用酶法檢測脂質含量,併用免疫印跡法分析胰島素受體和脂代謝相關蛋白在肝髒內的錶達. 結果 小鼠在註射含重組酶的腺病毒2d、4d和6d後,胰島素受體錶達依次降低30.50％ (P＜0.05)、60.12％ (P＜0.01)和99.54％ (P＜0.001),極低密度脂蛋白三酰甘油的分泌速率分彆降低20.43％ (P＜0.05)、33.63％(P＜0.05)和44.21％(P＜0.01);下調瞭固醇調控結閤蛋白1和脂肪痠閤成酶以及極低密度脂蛋白形成相關蛋白的錶達,但不影響新生載脂蛋白B100的分泌和肝髒脂質含量；而在含重組酶的腺病毒註射6d後新生載脂蛋白B48升高約35.07％ (P＜0.05).結論 肝髒胰島素信號急性缺失降低極低密度脂蛋白中三酰甘油的分泌速率併呈現時間依賴性,揭示小鼠肝髒分泌較小的極低密度脂蛋白顆粒,這種變化可能和肝髒內脂肪痠的閤成速度下降有關.
목적 분석간장조직특이성적이도소신호급성결실대재지단백B화삼선감유분비적영향. 방법 응용Cre-LoxP계통원리,재중조매기인적상유삽입간조직특이성백단백기인계동자,이차구건조직특이성표체적선병독.확증순화후적선병독경이도소수체Floxed소서미정맥주사,분별재주사2d、4d화6d후수집소서혈장,측정신생재지단백B화겁저밀도지단백삼선감유분비속솔,추제간장지질병용매법검측지질함량,병용면역인적법분석이도소수체화지대사상관단백재간장내적표체. 결과 소서재주사함중조매적선병독2d、4d화6d후,이도소수체표체의차강저30.50％ (P＜0.05)、60.12％ (P＜0.01)화99.54％ (P＜0.001),겁저밀도지단백삼선감유적분비속솔분별강저20.43％ (P＜0.05)、33.63％(P＜0.05)화44.21％(P＜0.01);하조료고순조공결합단백1화지방산합성매이급겁저밀도지단백형성상관단백적표체,단불영향신생재지단백B100적분비화간장지질함량；이재함중조매적선병독주사6d후신생재지단백B48승고약35.07％ (P＜0.05).결론 간장이도소신호급성결실강저겁저밀도지단백중삼선감유적분비속솔병정현시간의뢰성,게시소서간장분비교소적겁저밀도지단백과립,저충변화가능화간장내지방산적합성속도하강유관.
Objective To analyze the effects of acute depletion of liver-specific insulin signaling on secretion of apolipoprotein B (apoB) and triglyceride (TG).Methods Based on Cre-LoxP principle,a promoter of hepatic tissue specific albumin gene was inserted into upstream of the cre recombinase gene.Albumin-Cre adenovirus (Ad-CRE) and GFP adenovirus (Ad-GFP) were amplified in 293A cells and purified before intravenous administration to mice.After adenovirus infection for 2 days,4 days and 6 days,blood samples from mice were collected and hepatic tissues were frozen.The secretion rates of hepatic newly synthesized apoB and very low density lipoprotein (VLDL)-TG were determined by injection of Triton WR-1339.The levels of plasma cholesterol (TC) and TG were measured.The expressions of insulin receptor and other lipoprotein metabolism related proteins in hepatic tissues were analyzed by Western blot.Results After 2 d,4 d and 6 d of the Ad-CRE injection into mice,insulin receptor expression was reduced by 30.50％ (P＜0.05),60.12％ (P＜ 0.01) and 99.54％ (P＜0.001),and VLDL-TG secretion rate was decreased by 20.43％ (P＜0.05),33.63％ (P＜0.05) and 44.21％ (P＜0.01),respectively.Expressions of sterol regulatory binding proteins 1,fatty acid synthase,and the related proteins of VLDL-formation were decreased,but there were no changes in hepatic secretion of apoB100 and hepatic lipids.The hepatic secretion of apoB48 was increased by 35.07％ (P＜0.05) 6 d after Ad-CRE injection.Conclusions Acute depletion of hepatic insulin receptor might reduce VLDL-TG secretion in manner of time-dependent,and increase the assembly and secretion of smaller apoB-containing lipoproteins in mice liver,which is probably associated with decreased lipogenesis.
