5－羟甲基糠醛对皮质酮损伤型海马神经元P－synapsin Ⅰ蛋白表达的影响

5-羟甲基糠醛对皮质酮损伤型海马神经元P-synapsin Ⅰ蛋白表达的影响

张丽娜张天良1金国琴戴薇薇林梅2

(上海中医药大学基础医学院生化教研室，上海201203)

摘要〔〕目的观察熟地有效成分5-羟甲基糠醛对高浓度皮质酮致海马神经元损伤及学习记忆相关蛋白P-synapsinI蛋白表达的影响。方法用24 h新生大鼠制备原代海马神经细胞。培养第8天用皮质酮、RU38486、5-HMF处理细胞，将细胞分为正常组、模型组(1×10-4mol/L皮质酮)、拮抗剂组(10-4mol/L皮质酮+3 nmol/L RU38486)、5-羟甲基糠醛组(10-4mol/L 皮质酮+ 0.5 mg/L 5-HMF)。24 h后，通过SYTO13-PI双荧光染色法观察各组细胞的形态及存活情况，MTT法测定细胞活性，生化方法检测衰老特异性指标β-半乳糖苷酶活性， Western 印迹检测学习记忆相关分子P-synapsinⅠ的蛋白表达。结果与正常组相比，模型组细胞死亡较多，细胞活力下降，β-半乳糖苷酶活性升高，P-synapsinⅠ蛋白表达显著性降低；如上改变均被皮质酮受体拮抗剂RU38486逆转；一定浓度的5-羟甲基糠醛明显减少死细胞数量，提高细胞活力，降低β-半乳糖苷酶活性，提高模型细胞P-synapsinⅠ蛋白表达。结论0.5 mg/L 5-羟甲基糠醛可以保护大鼠海马神经细胞免遭高浓度皮质酮的损伤，通过调节学习记忆信号转导途径中的重要蛋白P-synapsinⅠ的蛋白表达，可能在延缓学习记忆功能退化中发挥作用。

关键词〔〕5-羟甲基糠醛；海马神经元；皮质酮；P-synapsinⅠ；熟地

中图分类号〔〕R966〔文献标识码〕A〔

基金项目：国家自然科学

Protein expression of P-synapsin Ⅰ following 5-HMF of SHUDI treatment in hippocampal neurons with corticosterone injury

ZHANG Li-Na,ZHANG Tian-Liang,JIN Guo-Qin,etal.ss

Department of Biochemistry,College of Basic Medicine,Shanghai University of TCM,Shanghai 201203,China

Abstract【】ObjectiveTo observe the effect of 5-HMF in SHUDI to P-synapsin Ⅰ protein expression in injured hippocampal neurons by high dose of corticosterone(CORT). MethodsHippocampal neurons were cultured from newborn (24 h) rats. On the eighth day the neurons were treated with CORT，RU38486 and 5-HMF. The cells were divided into control,model (containing 10-4mol/L CORT),RU38486 (containing 10-4 mol/L CORT+3 nmol/L RU38486) and 5-HMF (containing 10-4 mol/L CORT+0.5 mg/L 5-HMF) groups. 24 hours later，SYTO13-PI double-fluorescence staining,MTT assay,β-galactosidase assay and Western blot were carried out. ResultsCompared with the control group,both the cell survival rate and cell viability were decreased markedly (P<0.05),β-galactosidase activity was increased markedly(P<0.05) and P-synapsin Ⅰ protein expression was significantly decreased in model group. 5-HMF or RU38486 could invert above changes. Conclusions0.5 mg/L 5-HMF in SHUDI could significantly prevent the hippocampus neuron from injury. 5-HMF could change P-synapsin Ⅰ protein expression in injured hippocampal neurons by high dose of CORT to prevent and treat learning and memory impairment probably.

【Key words】5-HMF; Hippocampal neuron; Corticosterone(CORT); P-synapsin Ⅰ; SHUDI

1十堰市第十六中学2上海中医药大学教学实验中心

第一作者：张丽娜(1980-)，女，在读博士，讲师，主要从事中西医结合基础(生化) 研究。

补肾滋阴药能有效改善老年学习记忆减退〔1〕。……