CAJ | 학술논문

目的通过体外培养干预,研究激活Wnt信号通路对小鼠NIT-1胰岛细胞中PPARγ及葡萄糖激酶(Glucokinase,GK)表达的影响,探讨Wnt信号通路与PPARγ在胰岛细胞中的对话.方法重组Wnt3a蛋白干预体外培养的小鼠NIT-1胰岛细胞,激活Wnt信号通路,荧光定量PCR及Western印迹方法比较PPARγ的mRNA及蛋白水平,荧光定量PCR方法比较GK的mRNA水平.结果 Wnt3a干预组细胞的PPARγ及GK的mRNA水平均较对照组增加41.2％和65.0％ (P＜0.01),PPARγ蛋白表达亦明显增加(＜0.01).予dickkopf 1阻断Wnt信号通路,PPARγ与GK的表达较Wnt3a干预组降低,激活Wnt通路同时以wortmannin阻断磷酸肌醇3激酶(PI3K)通路,PPARγ与GK的的表达亦较单纯Wnt3a干预组降低.同时阻断Wnt及PI3K通路时,PPARγ的蛋白水平较Wnt3a干预组下降的更为显著,且较单独阻断Wnt或PI3K信号通路时更明显.结论激活Wnt信号通路,能够上调胰岛细胞PPARγ及GK的表达,且作用部分依赖于PI3K通路.
목적 통과체외배양간예,연구격활Wnt신호통로대소서NIT-1이도세포중PPARγ급포도당격매(Glucokinase,GK)표체적영향,탐토Wnt신호통로여PPARγ재이도세포중적대화.방법 중조Wnt3a단백간예체외배양적소서NIT-1이도세포,격활Wnt신호통로,형광정량PCR급Western인적방법비교PPARγ적mRNA급단백수평,형광정량PCR방법비교GK적mRNA수평.결과 Wnt3a간예조세포적PPARγ급GK적mRNA수평균교대조조증가41.2％화65.0％ (P＜0.01),PPARγ단백표체역명현증가(＜0.01).여dickkopf 1조단Wnt신호통로,PPARγ여GK적표체교Wnt3a간예조강저,격활Wnt통로동시이wortmannin조단린산기순3격매(PI3K)통로,PPARγ여GK적적표체역교단순Wnt3a간예조강저.동시조단Wnt급PI3K통로시,PPARγ적단백수평교Wnt3a간예조하강적경위현저,차교단독조단Wnt혹PI3K신호통로시경명현.결론 격활Wnt신호통로,능구상조이도세포PPARγ급GK적표체,차작용부분의뢰우PI3K통로.
Objective To investigate the expression of peroxisome proliferator-activated rec eptor γ (PPARγ) and glucokinase (GK) induced by Wnt signaling pathway in mice NIT-1 β-cells,and to explore the interaction between PPARγ and Wnt signaling pathways.Methods Recombinant Wnt3a protein was applied to NIT-1 beta-cells to activate Wnt signaling pathway.The expression of PPARγ was determined by real-time PCR and Western blotting.The expression of GK was determined by real time PCR.Results Wnt3a rapidly activated Wnt/β-catenin/TCF signaling pathway,and increased PPARγ and GK mRNA expression by 41.2％ and 65.0％ in NIT-1cells,with PPARγ protein expression increasing by 97.8％ (P＜0.01).These effects were abrogated by Wnt and PIK3 inhibitors,dickkopf 1 and wortmannin treatment (P＜ 0.01).Conclusions PPARγ and GK can be upregulated by Wnt singnaling,and the effects might partially be PI3K-dependent.