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目的分析wnt3a蛋白对小鼠胰腺beta细胞株增殖的影响。方法培养MIN6细胞株,并用不同浓度的wnt3a蛋白或者相同浓度wnt3a蛋白在不同时间点刺激MIN6细胞,用Western blot的方法检测MIN6细胞内的信号分子改变。培养MIN6细胞株,并用wnt3a蛋白刺激,分别在第0、1、2、3、4 d分裂并数细胞个数,比较有wnt3a刺激组与无wnt3a刺激组细胞个数有无区别。结果不同浓度wnt3a刺激,对于MIN6细胞内beta-catenin以及Pitx2的表达有影响,当wnt3a浓度为1:8时,beta-catenin以及Pitx2的表达最高。用1:8的wnt3a蛋白浓度刺激MIN6细胞,分别设置0 min、10 min、30 min、1 h、3 h、6 h、12 h、24 h、48 h。结果显示6 h时,beta-catenin表达量最高。比较控制组与wnt3a刺激组细胞增殖的区别,wnt3a蛋白刺激组细胞增殖程度明显高于控制组。结论wnt3a可以促进胰腺beta细胞的增殖,表明wnt信号通路对于胰腺beta细胞有重要的调控作用。
Objective To analyze the effect of wnt3a protein on the proliferation of mouse pancreatic beta cell line. METHODS MIN6 cells were cultured and the MIN6 cells were stimulated with different concentrations of wnt3a protein or the same concentration of wnt3a protein at different time points. The signal molecules in MIN6 cells were detected by Western blot. MIN6 cell lines were cultured and stimulated with wnt3a protein. The cells were divided at 0, 1, 2, 3, and 4 days respectively to count the number of cells. Compare the number of cells in wnt3a-stimulated group and non-wnt3a-stimulated group. Results Different concentrations of wnt3a stimulated the expression of beta-catenin and Pitx2 in MIN6 cells. When the concentration of wnt3a was 1: 8, the expression of beta-catenin and Pitx2 were the highest. MIN6 cells were stimulated with 1: 8 wnt3a for 0 min, 10 min, 30 min, 1 h, 3 h, 6 h, 12 h, 24 h, 48 h, respectively. The results showed that 6 h, the highest expression of beta-catenin. The difference between the control group and wnt3a stimulation group cell proliferation, Wnt3a protein stimulation group cell proliferation was significantly higher than the control group. Conclusion wnt3a can promote the proliferation of pancreatic beta cells, indicating that wnt signaling pathway plays an important regulatory role in pancreatic beta cells.