滋补脾阴方药含药血清对β淀粉样蛋白损伤原代培养大鼠海马神经元的保护作用及机制研究(英文)  

Protective effect of spleen-yin-nourishing recipe on amyloid β-peptide-induced damage of primarily cultured rat hippocampal neurons and its mechanism

作　　者：战丽彬[1] 牛新萍[2] 隋华[3] 宫晓洋[4] 

机构地区：[1]大连医科大学附属第二医院中医科,辽宁大连116023 [2]大连市第二人民医院干部病房,辽宁大连116011 [3]大连医科大学病理生理学教研室,辽宁大连116044 [4]大连医科大学附属第一医院中医科,辽宁大连116021

出　　处：《中西医结合学报》2009年第3期242-248,共7页Journal of Chinese Integrative Medicine

基　　金：国家自然科学基金资助项目(No.30472255)

摘　　要：目的:观察β淀粉样蛋白(amyloid β-peptide,Aβ)神经毒性、血清诱导激酶(serum-inducible kinase,SNK)-树突棘相关Rap特异性GTP酶活化蛋白(spine-associated Rap guanosine triphosphatase activating protein,SPAR)途径、N-甲基-D-门冬氨酸受体(N-methyl-D-aspartate receptor,NMDAR)三者的关系,探讨滋补脾阴方药保护Aβ损伤原代培养大鼠海马神经元的作用机制。方法:将干粉Aβ1-40配制成溶液,在37℃恒温箱中孵育72h,获得聚集态纤维状Aβ1-40。原代培养大鼠海马神经元,以血清药理学方法制备滋补脾阴方药含药血清,建立Aβ1-40(5μmol/L)损伤神经元模型,滋补脾阴方药含药血清为干预组,采用逆转录聚合酶链式反应方法观察SNK、SPAR及NMDAR亚基NR1、NR2A、NR2B mRNA表达。结果:与对照组相比,神经元暴露于Aβ1-40(5μmol/L)2h后,SNKmRNA表达上调,SPAR、NR1、NR2A和NR2B mRNA表达下调,差异有统计学意义(P<0.01,P<0.05)。与Aβ1-40(5μmol/L)组比较,各浓度滋补脾阴方药含药血清组SNKmRNA表达下调,SPAR、NR1、NR2A和NR2B mRNA表达上调,以2%滋补脾阴方药含药血清效果最显著(P<0.05)。结论:Aβ引起神经元损伤的过程与SNK-SPAR途径和NMDAR相关;滋补脾阴方药对Aβ损伤神经元有保护作用,其保护机制可能与调节NMDAR表达,阻断SNK-SPAR途径有关。Objective： To observe the relationship among amyloid β-peptide （Aβ）-induced neurotoxicity, serum-inducible kinase （SNK）-spine-associated Rap guanosine triphosphatase activating protein （SPAR） pathway and N-methyl-D-aspartate receptor （NMDAR）, and to explore the mechanism of the protective effect of spleen-yin nourishing recipe （Zibu Piyin Recipe, ZBPYR） in hippocampal neurons against Aβ-induced neurotoxicity. Methods： The Aβ1-40 powder was dissolved in 1× PBS and incubated at 37℃, and then aggregated fibrillar Aβ1-40 was obtained 72 h later. We used rat primary hippocampal neurons as cell model. ZBPYR-containing serum was gained by the method of serum pharmacology. ZBPYR-containing serum was added to the culture I h before Aβ1-40 （5 μmol/L） exposure. Cells were harvested 2 h after Aβ1-40 exposure for total RNA extracting. Then the mRNA expression levels of SNK, SPAR and NMDAR subunits NR1, NR2A and NR2B were detected by reverse transcription-polymerase chain reaction （RT-PCR）. Results： After 2-hour Aβ1- 40 exposure, we found that the expression level of SNK mRNA was up-regulated and the expression levels of SPAR, NR1, NR2A and NR2B mRNAs were down-regulated in hippocampal neurons as compared with control group （P〈0.01, P〈0.05）. While with ZBPYR-containing serum pretreatment, the expression level of SNK mRNA was down-regulated and the levels of SPAR, NR1, NRgA and NR2B were upregulated as compared with Aβ1-40 exposure, and 2 % ZBPYR-containing serum showed the best effect （P〈0.05）. Conclusion： Aβ-induced neurotoxicity was related to SNK-SPAR pathway and NMDAR; ZBPYR-containing serum can protect neurons from Aβ-induced neurotoxicity, and this protective effect may be performed by regulating the expression of NMDAR and blocking of the SNK-SPAR pathway.

关 键 词：阿尔茨海默病 滋补脾阴方药 淀粉样Β蛋白 血清诱导激酶 树突棘相关Rap特异性GTP酶活化蛋白 受体 N-甲基-D天冬氨酸 突触 

分 类 号：R285.5[医药卫生—中药学]