摘要
目的 :建立D -半乳糖致原代培养神经元损伤模型 ,为筛选益智药物提供药效学研究方法。方法 :在原代培养神经元的第 6d ,加入 5 0mmol/LD -半乳糖作用 72h ,倒置显微镜下观察神经元的生长发育形态 ,MTT法在酶联免疫检测仪上检测神经元代谢率 ,PI染色在流式细胞仪上检测神经元凋亡率 ,HE染色观察神经元形态、形态计量神经突起密度 ,RT -PCR检测醛糖还原酶信使RNA(AR -mRNA)含量。比较正常对照神经元和受D -gal攻击神经元之间的差别。结果 :受D -半乳糖攻击神经元的生长发育显著迟缓 ;代谢率从 0 76 2± 0 0 30 (n =33)降低到0 5 4 3± 0 0 6 4 (n =11) ,P <0 0 1;凋亡率从 0 0 6 0± 0 0 2 9(n =19)增高到 0 35 6± 0 2 15 (n =19) ,P <0 0 1;出现变性坏死的形态变化 ,神经突起密度从 0 5 5 7± 0 0 4 2 (n =10 )降低到 0 4 6 8± 0 0 33(n =10 ) ,P <0 0 1;无AR -mRNA的表达。结论 :用D -半乳糖建立的原代培养神经元损伤模型稳定、观察指标敏感 ,可用于老年性痴呆等神经系统疾病的研究。
AIM: To establish a model of primary cultured neuron injury induced by D-galactose for the research in Alzheimer's disease. METHODS: Primary rat neurons cultured for 6 days were exposed to 50 mmol D-galactose for 72 h. The neural growth and neurite density were observed with HE stain and microscope, the neural metabolism rate and apoptosis rate were examined with MTT, immuno-enzyme assay and flow cytometry, respectively, and the aldose reductase mRNA expression was also detected with RT-PCR. RESULTS: The neural growth and development in neurons treated with D-galactose was retarded, the neural metabolism rate decreased from 0.762±0 030( n= 33) to 0 543±0 064( n= 11)( P <0 01 ), and the neural apoptosis rate in injured neurons increased from 0 060±0 029 ( n= 19) to 0 356±0 215( n= 19), P <0 01 In addition, the neurite density in neurons treated with D-galactose decreased from 0 557±0 0422( n= 10) to 0 468±0 0330( n= 10)( P <0 01), the significant neural degeneration and necrosis were found. There is no aldose reductase mRNA expression in the neurons. CONCLUSION: A stable neuron injury model was established with 50 mmol/L D-galactose for 72 h, and it may be an useful tool for Alzheimer's disease research.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2002年第7期794-797,共4页
Chinese Journal of Pathophysiology
