Different Species Of Animal Liver Microsomal CYP450Enzymes Gelsemium Metabolism Impact

Purpose Gelsemium is a world-famous poisonous plants, which was described in herbal contribution of past dynasties, and has a long history to use. It was believed to have effect of qufeng challenging、sanjie and relieving pain. Modern research has shown Gelsemium roots, stems, leaves are highly toxic, in order to more effectively and security use of the alkaloide, we have carried out studies of the median lethal dose. Koumine as the highest alkaloid content Gelsemium plants, as the object of study, pigs, sheep and mouse liver microsomal comparative study of their metabolic characterization revealed part of its liver detoxification mechanisms. Swine, sheep, mice Gelsemium metabolism there is a different place, so we use the probe method study pigs, sheep, mice, and after Gelsemium induced cytochrome P450activity similarities and differences. Method Fujian Gelsemium roots first pulverized, and then with95%athanol extraction, recovery of ethanol under reduced pressure, plus2%hydrochloric acid, and then extracted with chloroform, the concentrated ammonia alkaline water layer, extracted with chloroform, got Gelsemium roots alkaloids. Based on the pre-test results of40mice were divided into four groups of10each, in accordance with the5.5mg/kg,4.4mg/kg,3.52mg/kg,2.82mg/kg dose disposable fed mice observed within24h situation. By recording mouse clinical symptoms and death situation, the acute toxicity observed from Fujian Gelsemium root total alkaloids and Sun modified Karber method LD50and its95%confidence interval. This topic using the Bradford method and Omura Determination of mice,pigs and sheep liver microsomal protein and CYP450contant were measured, using high performance liquid chromatography, chromatographic column:Yilite hypersil ODS colimn,(4.6mm×150mm,5μm), mobile phase:acetonitrile:0.1%triethylamine, the volume ratio of20:80; flow rate:1.0mL/min;column temperature:30℃; UV detection wavelength:254nm; injection volumn:20μL. Swine, sheep, mice Gelsemium metabolism there is a different place, so we use the probe method study pigs, sheep, mice, and after Gelsemium induced cytochrome P450activity similarities and differences. Rerult After the experimental study Fujian Gelsemium root total alkaloids The LD50after oral administration of4.12mg/kg,95%confidence interval3.76-4.52mg/kg. mouse,pig,sheep liver microsomal protein content6.973mg/mL,24.805mg/mL,28.200mg/mL, CYP450 content were0.379nmol/mg,0.798nmol/mg,0.847nmol/mg. In this chromatographic conditions, swine and sheep have metabolic koumine, mice no significant metabolism. The same time, we pig goat liver microsomal metabolism of koumine optimal conditions examined, the study found koumine optimal metabolism in the liver microsomes in vitro reaction system conditions:the incubation time of90min, liver microsomes the protein concentration of2.5mg/mL, the kpumine concentration of500μg/mL; koumine in the goat liver microsomal metabolism optimal vitro reaction system conditions:the incubation time of70min, hepatic mircrosomal protein concentration of3.0mg/mL, koumine concentration of500μg/mL. Ultimately through experiments with Gelsemium alkaloids speculate CYP2A6enzyme metabolism (degradation, parcels, etc.) have greater relevance. Conclusion Investigation probe method using pigs, sheep, mice, and after Oncorhynchus induced cytochrome P450activity so as to initially revealed the similarities and differences between the three mechanisms of the similarities and differences, and ultimately through experiments speculate CYP2A6enzyme metabolism of Gelsemium alkaloids (degradation, parcels, etc.) have greater relevance.