激活Wnt通路促进NIT-1胰岛β细胞PPARγ表达的研究

发布时间：2018-01-10 17:16

本文关键词：激活Wnt通路促进NIT-1胰岛β细胞PPARγ表达的研究　出处：《华中科技大学》2011年硕士论文　论文类型：学位论文

更多相关文章： Wnt PI3K PPARγ β-catenin TCF7L2 Akt GK

【摘要】：目的:通过体外培养干预小鼠NIT-1胰岛β细胞,观察Wnt信号通路对过氧化物酶体增生物激活受体γ(peroxisome proliferator-activated receptorγ,PPARγ)及葡萄糖激酶(glucokinase,GK)表达的调节作用,并探讨Wnt信号通路与PPARγ在β细胞中的复杂对话。方法:重组Wnt3a蛋白干预体外培养的小鼠NIT-1胰岛β细胞,荧光定量PCR及Western Blot方法检测PPARγmRNA及蛋白水平较对照组的变化,并给予Wnt通路阻断剂dickkopf 1(DKK1)及PI3K/Akt通路阻断剂wortmannin(Wort),比较PPARγ的表达量的改变。PCR法检测各干预组(对照组、Wnt3a组、Wnt3a+DKK1组、Wnt3a+Wort组)GK mRNA的表达。结果:细胞Wnt信号通路激活,PPARγmRNA水平较对照组增加了41.2%±7.8%(p0.05),蛋白表达较对照组增加了97.8%±29%(p0.05),GK的mRNA水平较对照组增加了65%±14.8%(p0.05)。DKK1阻断Wnt信号通路后,明显抑制了PPARγ与GK的mRNA表达,PPARγ的蛋白水平亦较Wnt3a干预组降低了36.2%±7.7%(p0.05),激活Wnt通路同时以Wort阻断PI3K通路,发现PPARγ与GK的mRNA水平下降,PPARγ的蛋白水平亦较单纯Wnt3a干预组降低了28.1%±3.8%(p0.05)。同时阻断Wnt及PI3K通路时,发现PPARγ的蛋白水平下降的更为显著,较Wnt3a组降低83.4%±4.9%(p0.05),作用较单阻断Wnt或PI3K信号通路时更明显(p0.05)。结论:1.在小鼠NIT-1胰岛β细胞,激活Wnt信号通路,能够上调PPARγ及GK的表达,影响细胞胰岛素分泌功能;2. Wnt3a对NIT-1细胞PPARγ的刺激作用部分依赖于PI3K/Akt通路的作用。
[Abstract]:Objective: to interfere with NIT-1 islet 尾 cells in vitro. To observe the effect of Wnt signaling pathway on peroxisome proliferator-activated receptor 纬. The expression of PPAR 纬 and glucokinase G K) was regulated, and the complex dialogue between Wnt signaling pathway and PPAR 纬 in 尾 cells was discussed. Methods: recombinant Wnt3a protein interfered with NIT-1 islet 尾 cells cultured in vitro. Fluorescence quantitative PCR and Western Blot were used to detect the changes of PPAR 纬 mRNA and protein levels compared with the control group. Wnt pathway blocker dickkopf 1 DKK1 and PI3K/Akt pathway blocker wortmanning Wort were given. To compare the change of PPAR 纬 expression. PCR method was used to detect Wnt3a DKK1 in each intervention group (control group Wnt3a group). Expression of GK mRNA in Wnt3a Wort group. Results: compared with the control group, the level of PPAR- 纬 mRNA activated by Wnt signaling pathway increased by 41.2% 卤7.8% (p0.05). Compared with the control group, the expression of protein increased by 97.8% 卤29% (p0.05). Compared with the control group, the mRNA level of GK increased by 65% 卤14.8. after blocking the Wnt signaling pathway, the mRNA expression of PPAR 纬 and GK was significantly inhibited by DKK1. The protein level of PPAR 纬 was also lower than that of Wnt3a intervention group (36.2% 卤7.7g / ml, p 0.05). The activation of Wnt pathway and the blocking of PI3K pathway by Wort were also observed. The mRNA levels of PPAR 纬 and GK decreased. The protein level of PPAR 纬 was also decreased by 28.1% 卤3.8and p0.05 compared with that in the Wnt3a intervention group. At the same time, the Wnt and PI3K pathways were blocked. It was found that the protein level of PPAR 纬 was significantly lower than that of Wnt3a group (83.4% 卤4.9 vs 0.05). The effect of P0. 05 was more obvious than that of single blocking of Wnt or PI3K signaling pathway. Conclusion 1. The activation of Wnt signaling pathway in mouse NIT-1 islet 尾 cells can up-regulate the expression of PPAR 纬 and GK and affect the insulin secretion function of the cells. 2. The stimulating effect of Wnt3a on PPAR 纬 in NIT-1 cells was partly dependent on the effect of PI3K/Akt pathway.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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