摘要
目的：通过考察芦荟大黄素在大鼠肝微粒体外代谢的主要条件,研究CYP450酶对芦荟大黄素代谢的影响。方法：随机将12只SD雄性大鼠分为空白对照组和苯巴比妥钠组,对空白对照组（0.5%CMC-Na溶液,10 ml/kg,ig）和PB组（苯巴比妥钠,75.0 mg/kg,ip）进行大鼠肝微粒体的诱导和制备,再将芦荟大黄素与大鼠肝微粒体进行体外孵育代谢,-80℃终止反应,三氯甲烷提取样品。用高效液相-荧光检测器（HPLC-FLD）测定芦荟大黄素含量,流动相为甲醇：0.1%磷酸水（70：30）,激发波长435nm,发射波长515nm。结果：测定方法专属性,回收率高,日内和日间精密度良好,稳定性好满足生物样品检测要求。以芦荟大黄素代谢消失率计,在肝微粒体蛋白浓度为0~1.0mg/ml范围内成线性增加趋势,1.0~2.5mg/ml趋势不明显;孵化时间0~30min成线性增加,且高于60%。芦荟大黄素代谢消失率随其浓度的增加呈线性增加趋势,达到一定浓度后成降低趋势。芦荟大黄素低、中、高浓度在PB、CMC-Na肝微粒中代谢消失率比较均为PPB-CMC-Na〈0.01。结论：芦荟大黄素在大鼠肝微粒体中的体外代谢具有酶促动力学特征,CYP450酶对芦荟大黄素的代谢起促进作用,苯巴比妥钠诱导的CYP450亚型酶参与了芦荟大黄素的代谢。
Objective： To study liver microsomal eytochrome P450 in rats on metabolism of aloe-emodin by studying the main conditions of metabolism of aloe-emodin in liver microsomal. Methods： 12 SD male rats were randomly divided into blank control group and PB group , the blank control group （0.5% CMC - Na solution, 10 ml/kg, ig） and PB （phenobarbital sodium, 75.0 mg/kg, ip） were carried out induction and preparation of rat liver microsomal. Aloe-emodin with liver microsomes were incubated and metabolize in vitro, -80℃ terminating the reaction. The samples were extracted by cc13 and determinated of the remaining Alce-emodin by HPLC-FD, with the mobile phase consisting of methanol-0. 1% phosphoric acid water （70：30）. The detection wavelength was set to excitation wavelength hex435nm and emission wavelength kex515nm. Results： The method had properties, high recoveries, good precision at intraday and day and good stability that meet the testing requirements of biological samples. It counted by the metabolic disappearance rate of aloe-emodin. The linear ranges of liver microsome protein concentration were 0 - 1.0mg/ml with increasing trend, 1.0 - 2.5mg/ml with no obvious trend, The incubation time was Linear increasing trend in 0 - 30min, and higher than 60%. the metabolic disappearance rate of aloe-emodin increased linearly with the increase of the concentration, and reached a certain concentration with a decreasing trend. The metabolic disappearance rate of aloe-emodin low, medium and high concentrations in PB, CMC-Na liver microsomes comparisons showed PPB-CMC-Na 〈0.01. Conclusion： Invitro metabolism of alce-emodin in rat liver microsomes has enzymatic kinetics. CYP450 enzymes played a catalytic role for metabolism of alce-emodin. CYP450 isoforms induced by PB involved in the metabolism of aloe-emodin.
出处
《中药药理与临床》
CAS
CSCD
北大核心
2014年第1期48-51,共4页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金资助项目(81274179)
河南省高校青年骨干教师资助项目(2009GGJS-065)
河南省教育厅自然科学研究计划项目(2011A360007)
