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目的探讨抑制NgR的表达对抑制性微环境中神经细胞轴突生长的影响。方法提取大鼠脑髓鞘膜蛋白作为皮层神经元培养的底物。细胞分4组:常规培养(A)组,髓鞘膜蛋白(B)组,siRNA转染+髓鞘膜蛋白(C)组,无关序列转染+髓鞘膜蛋白(D)组。行NgR siRNA的转染,转染后每天行βⅢ-tubulin免疫荧光染色,测量每个细胞的最长突起长度。结果转染后初期(24h、48h),A组细胞生长最好[突起长度分别为(22.5±12.2)μm和(59.1±23.6)μm],C组次之[突起长度分别为(17.1±10.2)μm和(44.7±22.5)μm],B组和D组突起生长抑制明显[B组突起长度分别为(8.1±5.8)μm和(27.7±13.7)μm),D组(7.3±5.8)μm和(24.9±12.4)μm]。到96h,各组间差异趋于消失。结论髓鞘膜蛋白对皮层神经元突起的生长有明显抑制作用。抑制NgR的表达能在一定程度上减轻这种抑制作用。
Objective To investigate the effect of inhibiting the expression of NgR on the axonal growth of nerve cells in inhibitory microenvironment. Methods Rat brain myelin membrane protein was extracted as a substrate for cultured cortical neurons. The cells were divided into 4 groups: conventional culture (A) group, myelin membrane protein (B) group, siRNA transfection + myelin membrane protein (C) group, unrelated sequence transfected myelin membrane protein (D) group. Line NgR siRNA transfection, day after transfection β Ⅲ-tubulin immunofluorescence staining, measuring the longest length of each cell protrusion. Results The cells in group A showed the best cell growth at 24 h and 48 h after transfection (the lengths of the protuberances were (22.5 ± 12.2) μm and (59.1 ± 23.6) μm, respectively) ) (μm) and (44.7 ± 22.5) μm, respectively. The protrusion growth of group B and group D was significantly inhibited (the lengths of protrusions in group B were (8.1 ± 5.8) μm and (27.7 ± 13.7) μm, And (24.9 ± 12.4) μm]. By 96h, the differences among groups tended to disappear. Conclusion Myelin membrane protein has a significant inhibitory effect on the growth of neurites in cortex. Inhibition of NgR expression can reduce this inhibition to a certain extent.