目的观察糖基化终产物(AGEs)对胎鼠海马神经前体细胞(NPCs)增殖和分化的影响。方法分离孕14.5d胎鼠海马,有限稀释法进行单克隆体外培养,分为不同浓度AGEs-牛血清白蛋白(BSA)组和对照组。分别用无血清NPCs培养基和胎牛血清促其增殖和诱导分化。MTT法和免疫荧光双标法结合激光共聚焦成像分别观察不同浓度AGEs对NPCs增殖和分化的影响。结果AGEs-BSA(50-400)mg/L干预后第3天和第7天,吸光度(A570nm)显著降低(P<0.01);AGEs-BSA(50-400)mg/L干预7d后,NeuN阳性细胞率显著低于对照组(P<0.05)。结论糖基化终产物抑制胎鼠NPCs的增殖和向神经元的分化。
Objective To survey the effects of advanced glycosylation end products (AGEs) on proliferation and differentiation of neural precursor cells (NPCs) from embryonic rat hippoeampus. Methods Single cell suspension was prepared from the hippocampus of 14.5 days embryos rat. After formation of the primary neurospheres, single cell clone was obtained with limited dilution method. Then the single cell clone spheres were digested and induced to proliferate and differentiate using nonserum medium and bovine serum. The effects of varied concentrations of AGE-BSA on the proliferation and differentiation of NPCs were investigated by mononuclear cell direct cytotoxity (MTT) reduction assay and immumofluorescence laser scanning method, respectively. Results AGEs-BSA (50-400 mg/L) could decrease the absorbance (A570nm) on days 3 and 7 after intervention significantly (P〈 0. 01). Percentage of MAP2 positive cells was significantly lower after 7-day intervention of AGEs-BSA (50-400 mg/L) than that in group C (P〈0. 05). Conclusion AGEs inhibit the proliferation and differentiation of NPCs from embryonic rat hippocampus.