摘要
目的探讨从红车轴草分离的5种异黄酮芒柄花素(formononetin)、大豆黄素(daidzein)、毛蕊异黄酮(calycosin)、红车轴草素(pratensein)、德鸢尾素(irilone)对脂多糖(LPS)诱导的多巴胺能神经元损伤的保护作用。方法利用LPS建立多巴胺能神经元损伤的细胞模型,利用液闪测定法检测[3H]DA摄取力以评价多巴胺能神经元的功能,利用免疫细胞化学技术进行酪氨酸羟化酶(tyrosine hydroxylase,TH)染色观测多巴胺能神经元的形态和数目,利用免疫细胞化学技术进行OX-42染色观测小胶质细胞的形态和数目。NO测定依据Griess反应,TNF-α测定采用酶联免疫法,超氧自由基测定采用细胞色素C还原法。结果从红车轴草分离的5种异黄酮均以剂量依赖方式减轻大鼠中脑原代混合培养体系中LPS诱导的多巴胺摄取力的下降和多巴胺能神经元数目的下降。而且,这5种异黄酮还抑制中脑混合培养体系和小胶质细胞纯化体系中LPS诱导的小胶质细胞的活化和TNF-α、NO和超氧自由基的生成。并且,这5种异黄酮作用强弱的顺序依次为:pratensein>daidzein>calycosin>formononetin>irilone。结论从红车轴草分离的5种异黄酮对LPS诱导的多巴胺能神经元的损伤均具有不同程度的保护作用,且保护作用强弱的顺序依次为:pratensein>daidzein>calycosin>formononetin>irilone,抑制小胶质细胞的活化是其作用机制之一。
Aim To explore the protective effect of five isoflavones（formononetin,daidzein,calycosin,pratensein,irilone） from Trifolium pratense on lipopolysaccharide（LPS）-induced dopaminergic neurodegeneration.Methods An in vitro model of LPS-induced dopaminergic neurodegeneration was used to investigate the protective effect of five isoflavones from Trifolium pratense on dopaminergic neurons against LPS-induced neurotoxicity by measuring theDA uptake and counting of TH-immunoreactive cells.Microglia were visualized by staining for the CR3 complement receptor with monoclonal antibody OX-42.The production of nitric oxide（NO）was determined by measuring the accumulated level of nitrite in the culture supernatant with the Griess reagent,and the release of tumor necrosis factor-α（TNF-α）was measured with a rat TNF-α enzyme-linked immunosorbent assay kit.The production of superoxide was determined by measuring the superoxide dismutase（SOD）-inhibitable reduction of cytochrome C.Results All five isoflavones dose-dependently attenuated LPS-induced decrease in dopamine uptake and the number of dopaminergic neurons in rat mesencephalic neuron-glia cultures.Moreover,they also significantly inhibited LPS-induced activation of microglia and production of TNF-α,NO and superoxide in rat mesencephalic neuron-glia cultures and microglia-enriched cultures.In addition,the potency order of these five isoflavones was：pratenseindaidzeincalycosinformononetinirilone.Conclusions All five isoflavones from Trifolium pratense may protect dopaminergic neurons from LPS-induced injury and their effects in inhibiting microglia activation may be one of the mechanisms.Moreover,the rank order of protective potency of these five isoflavones is：pratenseindaidzeincalycosinformononetinirilone.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2011年第3期390-396,共7页
Chinese Pharmacological Bulletin
基金
合肥工业大学博士专项基金项目(No2010HGBZ0293)
