本文關鍵詞：激活Wnt通路促進NIT-1胰島β細胞PPARγ表達的研究　出處：《華中科技大學》2011年碩士論文　論文類型：學位論文

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【摘要】：目的:通過體外培養(yǎng)干預小鼠NIT-1胰島β細胞,觀察Wnt信號通路對過氧化物酶體增生物激活受體γ(peroxisome proliferator-activated receptorγ,PPARγ)及葡萄糖激酶(glucokinase,GK)表達的調(diào)節(jié)作用,并探討Wnt信號通路與PPARγ在β細胞中的復雜對話。 方法:重組Wnt3a蛋白干預體外培養(yǎng)的小鼠NIT-1胰島β細胞,熒光定量PCR及Western Blot方法檢測PPARγmRNA及蛋白水平較對照組的變化,并給予Wnt通路阻斷劑dickkopf 1(DKK1)及PI3K/Akt通路阻斷劑wortmannin(Wort),比較PPARγ的表達量的改變。PCR法檢測各干預組(對照組、Wnt3a組、Wnt3a+DKK1組、Wnt3a+Wort組)GK mRNA的表達。 結(jié)果:細胞Wnt信號通路激活,PPARγmRNA水平較對照組增加了41.2%±7.8%(p0.05),蛋白表達較對照組增加了97.8%±29%(p0.05),GK的mRNA水平較對照組增加了65%±14.8%(p0.05)。DKK1阻斷Wnt信號通路后,明顯抑制了PPARγ與GK的mRNA表達,PPARγ的蛋白水平亦較Wnt3a干預組降低了36.2%±7.7%(p0.05),激活Wnt通路同時以Wort阻斷PI3K通路,發(fā)現(xiàn)PPARγ與GK的mRNA水平下降,PPARγ的蛋白水平亦較單純Wnt3a干預組降低了28.1%±3.8%(p0.05)。同時阻斷Wnt及PI3K通路時,發(fā)現(xiàn)PPARγ的蛋白水平下降的更為顯著,較Wnt3a組降低83.4%±4.9%(p0.05),作用較單阻斷Wnt或PI3K信號通路時更明顯(p0.05)。 結(jié)論:1.在小鼠NIT-1胰島β細胞,激活Wnt信號通路,能夠上調(diào)PPARγ及GK的表達,影響細胞胰島素分泌功能;2. Wnt3a對NIT-1細胞PPARγ的刺激作用部分依賴于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 緯. The expression of PPAR 緯 and glucokinase G K) was regulated, and the complex dialogue between Wnt signaling pathway and PPAR 緯 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 緯 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 緯 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- 緯 mRNA activated by Wnt signaling pathway increased by 41.2% 鹵7.8% (p0.05). Compared with the control group, the expression of protein increased by 97.8% 鹵29% (p0.05). Compared with the control group, the mRNA level of GK increased by 65% 鹵14.8. after blocking the Wnt signaling pathway, the mRNA expression of PPAR 緯 and GK was significantly inhibited by DKK1. The protein level of PPAR 緯 was also lower than that of Wnt3a intervention group (36.2% 鹵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 緯 and GK decreased. The protein level of PPAR 緯 was also decreased by 28.1% 鹵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 緯 was significantly lower than that of Wnt3a group (83.4% 鹵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 緯 and GK and affect the insulin secretion function of the cells. 2. The stimulating effect of Wnt3a on PPAR 緯 in NIT-1 cells was partly dependent on the effect of PI3K/Akt pathway.
【學位授予單位】：華中科技大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R363

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相關博士學位論文 前1條

1 桂書彥;Wnt信號通路對NIT-1胰島β細胞的作用及機制[D];華中科技大學;2010年

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本文編號：1406107