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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: To study the effects of Huanglian Jiedu Decoction on improving brain ischemia and brain injury by comparing pharmacological effects of Huanglian Jiedu Decoction, The effective part of the role and its chemical composition, to provide the experimental basis for the party’s refined separation. Method: The extract of Huanglian Jiedu Decoction was decocted with water (part I). The aqueous extract was concentrated and extracted with petroleum ether, ethyl acetate and water saturated n-butanol gradient to give petroleum ether extract (part II), acetic acid Ethyl ester extract (part Ⅲ), water-saturated n-butanol extract (part Ⅳ), residual aqueous supernatant (part Ⅴ) and residual water layer (part Ⅵ) The contents of 6 kinds of index components in each extract were determined by RP-HPLC. The mouse PC-12 cells of adrenal medullary pheochromocytoma MTT assay was used to determine the effects of Huanglian Jiedu Decoction and its extracts on the hypoxia and hypoxia-induced hypoxia injury in PC-12 cells induced by low glucose and DMEM. Results: The content of each index component in site Ⅴ was high, and the proportions of 6 index components were similar to site Ⅰ. The site Ⅰ and Ⅴ had a significant protective effect on hypoxia and hypoxia injury of PC-12 cells. Conclusion: Part Ⅴ is the effective part of Huanglian Jiedu Decoction in improving cerebral ischemia and puzzling.