摘要
目的:基于磷酸二酯酶(PDE)抑制活性,建立清金化痰汤抗炎活性测定方法,补充与完善该经典名方的质量控制体系。方法:采用高效液相色谱法(HPLC)建立PDE活性测定方法,考察清金化痰汤抑制PDE活性的量效关系,流动相甲醇-0.5%乙酸水溶液(5∶95),检测波长254 nm。通过测定多批清金化痰汤水提物冻干粉对PDE的抑制率,以中和酶的活性国际单位U标示其生物活性。结果:选择4 U·m L^(-1)PDE溶液,在反应底物环磷酸腺苷(c AMP)浓度为50μmol·L^(-1)、反应时间为60 min时,酶促反应稳定。在此反应体系下,当清金化痰汤水提物冻干粉终质量浓度为0.11~3.0 g·L^(-1)时,其对PDE的抑制作用呈现浓度依赖性。确定清金化痰汤水提物冻干粉待检质量浓度1 g·L^(-1),在该质量浓度下对PDE呈显著且稳定抑制作用,抑制率>45%,即1 mg的清金化痰汤水提物冻干粉至少可中和1.8 U PDE的活性。结论:建立了基于PDE抑制活性的清金化痰汤生物活性评价方法,以PDE活性国际单位U表征了清金化痰汤的抗炎活性,可为中药复方生物活性测定研究提供新方法。
Objective Based on the inhibitory activity of phosphodiesterase (PDE), a method for determining the anti-inflammatory activity of Qingjin Huatantang was established to supplement and improve the quality control system of this famous classical formula.Method High performance liquid chromatography (HPLC) was used to determine the activity of PDE, and the dose-effect relationship of inhibiting PDE activity of Qingjin Huatantang was investigated. The mobile phase consisted of methanol-0.5% acetic acid aqueous solution (5∶95), and the detection wavelength was 254 nm. By measuring the PDE inhibition rate of multiple batches of Qingjin Huatantang water extract lyophilized powder, biological activity was marked with the activity of the neutralizing enzyme in the international unit U.Result When the concentration of reaction substrate (cyclic adenosine monophosphate) was 50 μmol·L^(-1) and the reaction time was 60 min, the enzymatic reaction was stable with 4 U·mL^(-1) of PDE. In this reaction system, when the concentration of Qingjin Huatantang water extract lyophilized powder was 0.11-3.0 g·L^(-1), the inhibitory effect of PDE showed a concentration-dependent relationship. It was determined that the concentration of Qingjin Huatantang water extract lyophilized powder to be tested was 1 g·L^(-1), which showed a significant and stable inhibitory effect on PDE, and the inhibitory rate was >45%, that is, 1 mg of Qingjin Huatantang water extract lyophilized powder could neutralize the activity of 1.8 U PDE at least.Conclusion This study establishes a biological activity evaluation method of Qingjin Huatantang based on the inhibitory activity of PDE, and the anti-inflammatory activity of Qingjin Huatantang is characterized by international unit U of PDE activity, which can provide a new method for the determination of biological activity of traditional Chinese medicine compounds.
作者
肖顺丽
孙正霄
刘陆
张琼玲
丁世兰
许静
廖福龙
游云
XIAO Shunli;SUN Zhengxiao;LIU Lu;ZHANG Qiongling;DING Shilan;XU Jing;LIAO Fulong;YOU Yun(Institute of Chinese Materia Media,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第15期15-20,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家“重大新药创制”科技重大专项(2018ZX09721004-006-003)。
关键词
清金化痰汤
水提物
经典名方
磷酸二酯酶(PDE)
高效液相色谱法(HPLC)
生物活性
质量控制
Qingjin Huatantang
water extract
famous classical formulas
phosphodiesterase(PDE)
high performance liquid chromatography(HPLC)
biological activity
quality control
