调心方中β淀粉样蛋白结合成分的初步探讨.docVIP

　　调心方中β淀粉样蛋白结合成分的初步探讨 作者：程敏, 冯琼, 钱淑文, 高慧, 朱粹青 【关键词】 β淀粉样蛋白; 调心方; 乳酸脱氢酶; 老年性痴呆 Objective: To explore yloid protEin (Aβ) binding elements in heart-beneficial recipe (HBR, a pound traditional Chinese herbal medicine), eliorate the cytotoxicity of Aβ. Methods: The extract of HBR and Aβ1-40 munoblotting. Affi-gel-Aβ1-40 ents from the extract of HBR ance liquid chromatography (HPLC). The assay of lactic dehydrogenase (LDH) release from the primary cultured rat cortex neurons and the Affi-gel-Aβ1-40 treated HBR serum. Results: Immunoblotting examination shoount of Aβ1-40 ents from the extract of HBR, eluted from Affi-gel-Aβ1-40 by glycine solution (pH=2.5), could be detected by HPLC-fluorescent detector system. The analysis of LDH release shool/L Aβ1-42 for 48 h caused a significant increase of LDH release in EIther a serum free or 10% serum contained culture condition (Plt;0.01). The rat HBR serum still maintained the ability to attenuate Aβ1-42 induced LDH release although the effect e. Conclusion: There are Aβ affinity ponents in HBR, ight be able to inhibit the cytotoxicity of Aβ. The results implied that the exploration of Aβ affinity elements from Chinese medicinal recipe er disease, might be an important direction in Alzheimer disease therapeutic research area. Keyyloid protein; heart-beneficial recipe; lactic dehydrogenase; Alzheimer disease 老年性痴呆（A1zheimer disease, AD）是一种与增龄相关的神经系统退行性疾病。随着社会人口的老龄化，其发病率呈上升趋势。β淀粉样蛋白（β-amyloid protein, Aβ）不仅是AD脑病理结构之一，即老年斑的主要组分，而且Aβ所介导的神经毒性在AD脑病理改变中起非常重要的作用，它可激活胶质细胞分泌多种细胞因子，诱导与神经纤维缠结相关的tau蛋白磷酸化，诱导神经元凋亡等［1］。因此，目前抗Aβ的神经毒性作用已成为防治AD研究的重要方向之一［2］。调心方的临床研究提示其有改善AD痴呆症状的作用［3，4］，而实验研究已经显示其可减少Aβ诱导的神经毒性［5-7］。本实验试图探索调心方中是否有直接与Aβ作用并抑制Aβ神经毒性的成分。 1 材料与方法 1.1 调心方提取物Aβ结合分析 1.1.1 调心方提取物与Aβ共沉淀并对沉淀物中的Aβ进行免疫印迹检测 调心方提取液，由党参、桂枝、茯苓、远志、石菖蒲等组成，用水提醇沉法制成相当于含生药7.4 g/ml的水溶液，成年人用量相当于生药2.48 g/kg体质量，由上海市中医老年医学研究所提供。不同浓度稀释的调心方1 ml加Aβ1-40 50 μg（吉尔生化上海有限公司）混匀后，于室温条件下在旋转混合仪（10 r/min）上作用2 h；12 000 r/min离心10 min。取沉淀加同量十二烷基硫酸钠（sodium dodecyl sulfate, SDS