Preliminary assay of beta-amyloid binding elements in heart-beneficial recipe / 中西医结合学报

ABSTRACT

OBJECTIVE: To explore whether there are beta-amyloid protein (Abeta) binding elements in heart-beneficial recipe (HBR, a compound traditional Chinese herbal medicine), which can ameliorate the cytotoxicity of Abeta. METHODS: The extract of HBR and Abeta(1-40) were co-precipitated, and the Abeta(1-40) in pellets was detected by immunoblotting. Affi-gel-Abeta(1-40) was constructed, and Affi-gel-Abeta(1-40) affinity elements from the extract of HBR were analyzed by high-performance liquid chromatography (HPLC). The assay of lactic dehydrogenase (LDH) release from the primary cultured rat cortex neurons was used to evaluate the cytotoxicity of Abeta(1-42), and the protection effects of the HBR serum and the Affi-gel-Abeta(1-40) treated HBR serum. RESULTS: Immunoblotting examination showed Abeta(1-40) could be co-precipitated with components of HBR following co-incubation, and the amount of Abeta(1-40) within pellets decreased when the HBR extract was diluted. Abeta(1-40) affinity elements from the extract of HBR, eluted from Affi-gel-Abeta(1-40) by glycine solution (pH=2.5), could be detected by HPLC-fluorescent detector system. The analysis of LDH release showed that exposure of neurons to 5 micromol/L Abeta(1-42) for 48 h caused a significant increase of LDH release in either a serum free or 10% serum contained culture condition (P<0.01). The rat HBR serum was able to suppress Abeta(1-42) induced LDH release (P<0.05), whereas Affi-gel-Abeta(1-40) treated HBR serum still maintained the ability to attenuate Abeta(1-42) induced LDH release although the effect was somewhat decreased compared with Affi-gel treated HBR serum. CONCLUSION: There are Abeta affinity components in HBR, which could not increase the Abeta cytotoxicity, but might be able to inhibit the cytotoxicity of Abeta. The results implied that the exploration of Abeta affinity elements from Chinese medicinal recipe which is effective for Alzheimer disease, might be an important direction in Alzheimer disease therapeutic research area.