目的:研究比较黄连解毒汤全方水提液体系、水提液体系经有机溶剂梯度萃取后不同极性部位中指标性成分的变化,通过药效学筛选,探讨黄连解毒汤改善脑缺血、益智作用的有效部位及其化学组成,为该方的精制分离提供实验依据。方法:黄连解毒汤全方经水煎煮得水提液(部位Ⅰ),水提液浓缩后以石油醚、乙酸乙酯和水饱和正丁醇梯度萃取,得到石油醚萃取物(部位Ⅱ)、乙酸乙酯萃取物(部位Ⅲ)、水饱和正丁醇萃取物(部位Ⅳ)、残留水层上清液(部位Ⅴ)、残留水层沉淀(部位Ⅵ),以栀子苷、黄芩苷、巴马汀、小檗碱、黄芩素和汉黄芩素6种成分为对照,应用RP-HPLC确定各萃取物中6种指标性成分的含量;选用鼠源肾上腺髓质嗜铬细胞瘤PC-12细胞活性跟踪筛选,MTT法测定黄连解毒汤及各萃取部位对低糖DMEM和氯化钴诱导的PC-12细胞缺糖缺氧损伤的影响。结果:部位Ⅴ中各指标性成分含量较高,且6种指标性成分的比例与部位Ⅰ相似;部位Ⅰ,Ⅴ对PC-12细胞的缺糖缺氧损伤保护作用明显。结论:部位Ⅴ为黄连解毒汤改善脑缺血、益智作用的有效部位。
Objective:This paper studies Chinese herbal compound of Huanglian Jiedu decoction and its extract systems by gradient extraction with three kinds of organic solvents and the changes in the indicative components in different polarity parts.Through pharmacodynamics screening,utility location and chemical composition which can improve cerebral ischemia and promote noopsychic function were approached,to provide the basis for refining separation.Method: Huanglian Jiedu decoction was extracted successively with water(partⅠ),ethyl acetate,water saturated butanol,the petroleum ether extract(part Ⅱ),ethyl acetate extract(part Ⅲ),butanol extract water saturated butanol extract(partⅣ),supernatant(partⅤ) and sediment(partⅥ) of residual water layer extract.Using RP-HPLC,and geniposide,baicalin,palmatine,berberine,baicalein and wogonin as the indicative components,each component in every extract system was detected.Finally,using PC12 cells,low-sugar of DMEM and cobalt chloride to made oxygen and glucose deprivation model,MTT was applied to detect the cell viability.Result: The quality of the indicative components in partⅤwas higher,having the similar ratio with part Ⅰ;part Ⅰ and part Ⅴ had the highest protective effect in the oxygen and glucose deprivation injured cells.Conclusion: PartⅤ is the effective fractions that can improve cerebral ischemia and noopsychic function.