血管内皮生长因子165对神经干细胞生长及分化的影响

Effect of VEGF165 on the growth and differentiation of hippocampal neural stem cells

目的观察血管内皮生长因子165(VEGF165)对大鼠海马神经干细胞(NSC)增殖和分化的影响。方法自新生1 d SD大鼠提取NSC,分为4组,A组加入正常培养基,B组加入含10%胎牛血清的DMEM/F12培养基,C组加入含20μg/L VEGF的正常培养基,D组加入含20μg/L VEGF和10%胎牛血清的DMEM/F12培养基,通过免疫荧光染色方法比较4组间Nestin、β-tubulinⅢ、GFAP三种蛋白的表达情况,并计算各组阳性细胞率,通过统计学分析软件SPSS 16.0,采用两独立样本间t检验方式,比较A组与C组、B组与D组、C组与D组间各染色阳性率差异,检测水准α=0.05,研究VEGF165对于NSC的增殖与分化的作用。结果 A组与C组、B组与D组、C组与D组间Nestin染色阳性率差异均有统计学意义(t=31.79、-68.89、21.33,P均<0.01),β-tubulinⅢ染色阳性率差异有统计学意义(t=-33.18、-99.78、30.61,P均<0.01)、GFAP免疫荧光染色阳性率差异有统计学意义(t=28.89、27.07、-22.59,P均<0.01)。结论 VEGF165是NSC增殖及分化的促进因素,且可能存在诱导NSC向神经元分化的作用。

Objective To observe the effective of vascular endothelial cell growth factor（VEGF-165） on the proliferation and differentiation of rat hippocampus neural stem ceils （NSC）. Methods Extracted NSC from newborn SD rats, and divided into four groups：Group A was added to normal culture medium;Group B added DMEM/F12 medium containing 10% fetal bovine serum; Group C added normal culture medium containing 20 ~g,/L VEGF; Group D added DMEM/F12 medium containing 20 ~g/L VEGF and the 10% FBS. Compare the expression of Nestin, 13-tubulin m and GFAP by immunofluorescence staining method among the four groups, and calculated positive cell rate of each group by Statistics analysis to study the role of VEGF165 for the proliferation and differentiation of NSC. Results The t value of immunofluorescence staining positive rate of Nestin between group A and C, group B and D, group C and D was 3 i. 79, - 68. 89, 21.33. The t value of ^-tubulin m was -33.18, -99. 78,30. 61. The t value of GFAP was 28.89,27.07, - 22. 59. All the value of P 〈 0. 01, and the difference was statistically significant. Conclusion VEGF165 is a promoting factor of the proliferation and differentiation of neural stem cells, and the effect that induced neural stem cells into neurons may exist.