中华内分泌代谢杂志
中華內分泌代謝雜誌
중화내분비대사잡지
CHINESE JOURNAL OF ENDOCRINOLOGY AND METABOLISM
2010年
2期
135-137
,共3页
线粒体融合素2基因%基因沉默%糖代谢%胰岛素抵抗
線粒體融閤素2基因%基因沉默%糖代謝%胰島素牴抗
선립체융합소2기인%기인침묵%당대사%이도소저항
Mitofusin-2 gene%Gene silencing%Glycometabolism%Insulin resistance
目的 研究RNA干扰(RNAi)介导线粒体融合素2(mitofusin-2,Mfn2)基因沉默对BALB/c小鼠葡萄糖代谢和胰岛素敏感性的影响.方法 构建带有绿色荧光蛋白(GFP)基因的Mfn2短发卡RNA质粒载体(Mfn2 shRNA)和阴性对照质粒载体(HK).44只BALB/c小鼠分为转染组(Mfn2)和阴性对照组.将含有75μg质粒溶液1.5 ml以流体力学法注入小鼠体内.质粒注射5 d后,采用腹腔葡萄糖耐量和胰岛素耐量试验观察Mfn2基因干扰对小鼠葡萄糖代谢和胰岛素敏感性的影响;采用氚标记葡萄糖(3-[~3H]-葡萄糖)通过尾静脉注射到小鼠体内,留取血标本,液体闪烁计数仪测定标本放射性浓度,计算小鼠肝脏葡萄糖生成率;Western印迹法检测肝脏组织中Mfn2蛋白表达.结果 与阴性对照组小鼠比较,Mfn2组小鼠肝脏组织Mfn2蛋白表达明显减少(8.05±0.15对8.56±0.01,P<0.05),空腹血糖升高[(6.95±0.83对4.68±0.29)mmol/L,P<0.05],胰岛素敏感性下降,肝脏葡萄糖生成增加[(49.43±16.31对24.91±4.07)μmol·kg~(-1)·min~(-1),P<0.05].结论 下调Mfn2基因表达使BALB/c小鼠葡萄糖代谢异常和胰岛素抵抗,Mfn2基因在维持体内葡萄糖代谢稳态中起重要作用.
目的 研究RNA榦擾(RNAi)介導線粒體融閤素2(mitofusin-2,Mfn2)基因沉默對BALB/c小鼠葡萄糖代謝和胰島素敏感性的影響.方法 構建帶有綠色熒光蛋白(GFP)基因的Mfn2短髮卡RNA質粒載體(Mfn2 shRNA)和陰性對照質粒載體(HK).44隻BALB/c小鼠分為轉染組(Mfn2)和陰性對照組.將含有75μg質粒溶液1.5 ml以流體力學法註入小鼠體內.質粒註射5 d後,採用腹腔葡萄糖耐量和胰島素耐量試驗觀察Mfn2基因榦擾對小鼠葡萄糖代謝和胰島素敏感性的影響;採用氚標記葡萄糖(3-[~3H]-葡萄糖)通過尾靜脈註射到小鼠體內,留取血標本,液體閃爍計數儀測定標本放射性濃度,計算小鼠肝髒葡萄糖生成率;Western印跡法檢測肝髒組織中Mfn2蛋白錶達.結果 與陰性對照組小鼠比較,Mfn2組小鼠肝髒組織Mfn2蛋白錶達明顯減少(8.05±0.15對8.56±0.01,P<0.05),空腹血糖升高[(6.95±0.83對4.68±0.29)mmol/L,P<0.05],胰島素敏感性下降,肝髒葡萄糖生成增加[(49.43±16.31對24.91±4.07)μmol·kg~(-1)·min~(-1),P<0.05].結論 下調Mfn2基因錶達使BALB/c小鼠葡萄糖代謝異常和胰島素牴抗,Mfn2基因在維持體內葡萄糖代謝穩態中起重要作用.
목적 연구RNA간우(RNAi)개도선립체융합소2(mitofusin-2,Mfn2)기인침묵대BALB/c소서포도당대사화이도소민감성적영향.방법 구건대유록색형광단백(GFP)기인적Mfn2단발잡RNA질립재체(Mfn2 shRNA)화음성대조질립재체(HK).44지BALB/c소서분위전염조(Mfn2)화음성대조조.장함유75μg질립용액1.5 ml이류체역학법주입소서체내.질립주사5 d후,채용복강포도당내량화이도소내량시험관찰Mfn2기인간우대소서포도당대사화이도소민감성적영향;채용천표기포도당(3-[~3H]-포도당)통과미정맥주사도소서체내,류취혈표본,액체섬삭계수의측정표본방사성농도,계산소서간장포도당생성솔;Western인적법검측간장조직중Mfn2단백표체.결과 여음성대조조소서비교,Mfn2조소서간장조직Mfn2단백표체명현감소(8.05±0.15대8.56±0.01,P<0.05),공복혈당승고[(6.95±0.83대4.68±0.29)mmol/L,P<0.05],이도소민감성하강,간장포도당생성증가[(49.43±16.31대24.91±4.07)μmol·kg~(-1)·min~(-1),P<0.05].결론 하조Mfn2기인표체사BALB/c소서포도당대사이상화이도소저항,Mfn2기인재유지체내포도당대사은태중기중요작용.
Objective To study the effects of RNA interference(RNAi)-mediated silencing of mitofusin-2 (Mfn2) gene on glycornetabolism and insulin resistance in BALB/c mice. Methods Mfn2 short hairpin RNA (shRNA) and negative control green fluorescent protein(GFP) -expressed plasmid vectors were constructed. 44 mice were randomly divided into transfection group (Mfn2) and negative control group (HK). 1.5 ml GFP-expressed plasmid(negative control or Mfn2 shRNA,75 μg for each mouse)was injected into the mice in 5 seconds through vena caudalis. Five days later, intraperitoneal glucose tolerance test (IPGTT) and intraperitoneal insulin tolerance test(IPITT)were performed to evaluate glycometabolism and insulin sensitivity. D-3-[3~H]-glucose in PBS buffer were injected via the tail vein. Blood samples were taken at specific time points. Radioactivity was measured in all samples with liquid scintillation counter. The rates of hepatic glucose production in vivo were calculated. Mfn2 protein expression in hepatic tissue was detected by Western blot. Results Compared with HK mice, the Mfn2 expressions of Mfn2 mice decreased markedly(8.05±0.15 vs 8.56±0.01 ,P<0.05). The fasted blood glucose leves [(6.95±0. 83 vs 4.68±0. 29) mmol/L,P<0. 05] in Mfn2 mice were higher than those in HK mice. The insulin sensitivity of Mfn2 mice decreased markedly compared with HK mice. The rate of hepatic glucose production was significantly elevated in Mfn2 mice [(49.53±16.31)μmol·kg~(-1)·min~(-1)],compared with negative control mice[(24.91±4.07)μmol·kg~(-1)·min~(-1),P<0.05].Conclusion The down-regulatd expression of Mfn2 induces glycometabolic disorder and insulin resistance in BALB/c mice. Mfn2 plays an important role in maintaining glucose homeostasis in vivo.
