无糖对体外培养的神经前体细胞损伤的影响

目的 :研究无糖处理对体外分离培养神经前体细胞 (neuralprecursorcells,NPCs)的存活、分裂及分化能力的影响。方法 :培养的第 4～ 6代新生大鼠海马区NPCs给予无糖培养基处理 6h后 ,分别于处理后第 0、1和 7天测定细胞MTT代谢率、台盼蓝染色细胞率、5′ 溴 脱氧尿嘧啶 (5′ bromo deoxyuridine,BrdU)参入率及分化细胞内胶质细胞纤维酸性蛋白、神经元特异性烯醇酶和半乳糖脑苷蛋白表达 ,判定NPCs存活、分裂和分化能力的改变。结果 :(1 )无糖处理后的不同时间点台盼蓝染色阳性细胞率在NPCs无糖组较NPCs对照组增加。无糖处理后即刻和 1d ,NPCs无糖组较神经元无糖组减少 ,7d时两者相近。 (2 )无糖损伤后 0d和 1d ,NPCs无糖组的MTT代谢率较NPCs对照组明显降低 (P <0 .0 5 ) ,7d时差异无显著性 (P >0 .0 5 )。在不同时间点 ,NPCs无糖组的MTT代谢率较神经元无糖组均明显增高 (P <0 .0 5 )。 (3)无糖处理后 ,NPCs无糖组的BrdU参入率与对照组相比 ,差异无显著性。 (4 )无糖对NPCs分化成为神经元、星形胶质细胞和少突胶质细胞的比例无显著性影响 (P >0 .0 5 )。结论 :无糖可以损伤NPCs ,对于NPCs的分裂和分化能力无直接的影响

Effect of glucose-free treatment on the neural precursor cells in vitro

OBJECTIVE: To investigate the survival, division, and differentiation of neural precursor cells (NPCs) following glucose-free treatment in vitro. METHODS: Cultured neonatal rat hippocampal NPCs were exposed to glucose-free media for 6 hours, and then were returned to regular media containing normal level glucose. At different time points after glucose-free treatment, trypan blue staining was used to determine cell viability, MTT assay to measure cell metabolic rate, 5'-bromo-deoxyuridine (BrdU) incorporation to determine the NPCs division, and immunocellulochemistry and western blot analysis to study the NPCs differentiation. RESULTS: (1) At different time points (0, 1, 7 d) after glucose-free treatment, NPCs viability decreased compared with NPCs control. On 0 and 1 d after glucose-free treatment, NPCs viability was higher than neurons viability. (2) On 0 and 1 d after glucose-free treatment, NPCs metabolic rates decreased significantly compared with NPCs control. On 0, 1, and 7 d after glucose-free treatment, NPCs metabolic rates were higher than that in neurons. (3) After glucose-free treatment, BrdU incorporation of NPCs was similar to NPCs control (P>0.05). (4) After glucose-free treatment, NPCs differentiation didn't change significantly (P>0.05). CONCLUSION: These findings demonstrated that glucose-free treatment impaired NPC's viability, but didn't affect the division and differentiation of NPCs.