波形蛋白影响PC12细胞神经分化的研究

Effect of Vimentin on Neural Differentiation of PC12 Cells

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 explore the function of vimentin on neural differentiation of PC12 cells. Methods Western blot was used to detect vimentin expression between PC12H and PC12L cells. Western blot was applied to detect vimentin expression of PC12 cells be- fore and after NGF treatment. Vimentin interference vector was constructed and the interference effect of vimentin expression was tested by Western blot. PC12H cells were transiently transfected with control vector, vimentin interference vector, respectively. After cultured for 0day, lday, 2days, 3days, we observed and photographed the morphology of cells. The neurite length and the population of neurite - branching cells were quantified by Image - pro plus software. Results Vimentin expression in PC12H cells was higher than PC12L cells （P = 0. 000）. Vimentin expression in PC12 cells was increased after NGF treatment and the change was more remarkable in PC12L cells （P = O. 000）. Vimentin interference vector had a noteworthy suppression effect on vimentin expression（P = 0. 000）. PC12H cells transi- ently transfected with vimentin interference vector had lower differentiation morphologies compared to control ones. The neurite length of cells was shorter and the population of neurite - branching cells was lower in interference vector group than control one, difference was sta- tistically significant（P = 0. 000）. Conclusion Vimentin can promote nueral differentiation of PC12 cells.