摘要
黄连解毒汤化学成分的研究及其治疗老年痴呆有效成分的筛选
黄连解毒汤是清热解毒的经典代表方剂,始出自《肘后备急方》名见《外台秘要》引崔氏方：“前军督护刘车者,得时疾三日已汗解,因饮酒复剧,苦烦干呕,口燥呻吟,错语不得卧,余思作此黄连解毒汤。”黄连解毒汤由黄连、黄芩、黄柏、栀子四味药材组成,主治一切实热火毒,三焦热盛之证。现代药理研究表明黄连解毒汤除了具有抗炎抑菌作用外,对肿瘤、脑缺血、糖尿病等疾病均具有良好效果,在日本,黄连解毒汤还用来治疗脑缺血后遗症等。因此,本文对黄连解毒汤进行了化学成分的研究并对其主要成分进行了老年性痴呆方面的筛选。
对黄连解毒汤的正丁醇部分进行分离后,采用各种色谱分离技术,分离得到11个化合物,结合化合物的理化性质,通过IR、NMR、UV和MS等多种分析手段,解析了其中11个化合物的结构。
运用DPPH (2,2-diphenyl-1-picryl-hydrazylhydrate)分光光度测定法对黄连解毒汤复方、单味药材、及其主要成分的抗氧化活性进行了检测。运用体外酶抑制剂的检测方法对黄连解毒汤中具有乙酰胆碱酯酶抑制活性的成分进行了筛选,结果发现：黄连解毒汤中具有抗氧化活性的成分为黄酮类成分,而生物碱类成分具有较好的乙酰胆碱酯酶抑制活性。
运用体外培养神经细胞的药理模型,对黄连解毒汤中具有神经保护作用的部位和成分进行了筛选比较,结果发现,黄连解毒汤主要成分均具有神经细胞保护作用的趋势,但没有统计学显著性。复合成分具有明显的细胞保护作用,可以减轻氧化、缺氧对PC12细胞、原代神经元的损伤,并且呈剂量依赖性,值得深入研究和开发。
Huanglianjiedu Decoction is a traditional Chinese herbal prescription, which was firstly recorded in A Handbook of Prescriptions for Emergencies, by Ge Hong in the Eastern Jin Dynasty in the 3rd Century A.D. It is a heat clearing and detoxicating formula, which is used to clear pathogenic fire. The formula consists of four crude drugs:Rhizoma coptidis, Radix scutellariae, Cortex phellodendri and Fructus gardeniae and was widely used both in China and Japan.
Nowadays, there are a lot of reaserches on Huanglianjiedu Decoction and it's effects on senile dementia. But only little research focuses on the active compounds.
In this study, ten major compounds were isolated from the n-butanol fraction of the water extract of Huanglianjiedu Decoction, the structures of these compounds were studied by chemical and spectral analysis, such as IR、NMR、UV and MS.
The acetylcholinesterase inhibitory and antioxidant activity effect of the formula HLJDT, the single herbs and the major compounds of HLJDT were determined and compared by acetylcholinesterase inhibition and antioxidant assay. The formula HLJDT had a better antioxidant activity effect than the single herbs, except for Radix Scutellaria which is rich in polyphenolic compounds, the most active compound is baicalin; The results also show that the herb with the best effect on AChE inhibition is Coptidis Rhizoma and the pure compound is berberine; it can be concluded that alkaloid compounds have best effect the inhibitory activity of the formula and flavanoids compounds on the antioxidant activity.
Traumatic models using PC 12 cells and primary neurons cells treated with glutamate were applied to investigate the cell protective effect of HLJDT and its major compounds. The cell and chondriosome viability were analyzed by the JC-1 fluorescent method, the trypan blue exclusion method and MTT method. All of the single pure compounds do not show any protective effect, but the mixed pure compounds showed protective effect on PC 12 cell and cultured primary neurons treated by glutamate at various concentration from 1-0.1μg/ml in dose-depend manner. This results can be used to illuminate the function of complex prescription.
引文
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