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目的探究波形蛋白分子在PC12细胞神经分化中的作用。方法用免疫印迹方法检测PC12H和PC12L细胞中波形蛋白的表达情况。用免疫印迹方法检测PC12H细胞和PC12L细胞在NGF处理前后波形蛋白的表达情况。构建波形蛋白干扰质粒,用免疫印迹方法检测该质粒的干扰效果。分别将对照质粒、波形蛋白干扰质粒瞬时转染PC12H细胞,培养0、1、2、3天观察与记录细胞的分化情况,用Image-pro plus软件定量分析这两组细胞的平均神经突长度和含分化神经突细胞的百分比例。结果 PC12H细胞中波形蛋白的表达明显高于PC12L细胞(P=0.000)。NGF处理后PC12H和PC12L细胞中波形蛋白的表达量均增加,且在PC12L细胞中更明显(P=0.000)。波形蛋白干扰质粒能有效地抑制PC12H细胞中波形蛋白的表达(P=0.000)。瞬时转染波形蛋白干扰质粒能有效地抑制PC12H细胞的神经分化,定量分析结果显示干扰组细胞的平均神经突长度和含分化神经突细胞的百分比均明显小于对照组,差异均有统计学意义(P=0.000)。结论波形蛋白分子可以促进PC12细胞的神经分化。
Objective To investigate the role of vimentin in neural differentiation of PC12 cells. Methods Western blotting was used to detect the expression of vimentin in PC12H and PC12L cells. The expression of vimentin in PC12H cells and PC12L cells before and after NGF treatment was detected by Western blotting. Construction of vimentin interference plasmids, Western blotting method to detect the interference effect of the plasmid. The control plasmids and vimentin interference plasmids were transiently transfected into PC12H cells respectively. The cell differentiation was observed and recorded on day 0, day 1, day 2, day 3, and the average neurite length Examples of percentages of differentiated neuronal cells. Results The expression of vimentin in PC12H cells was significantly higher than that in PC12L cells (P = 0.000). The expression of vimentin in PC12H and PC12L cells increased after NGF treatment, and more significantly in PC12L cells (P = 0.000). The vimentin interference plasmid can effectively inhibit the expression of vimentin in PC12H cells (P = 0.000). Transient transfection of vimentin interference plasmids could effectively inhibit the neural differentiation of PC12H cells. Quantitative analysis showed that the average neurite length and the percentage of differentiated neuronal cells in the interfering cells were significantly smaller than those in the control group (P <0.05) P = 0.000). Conclusion Vimentin can promote neural differentiation of PC12 cells.