Research Distribution Of Radix Rubiae Active Fraction In Living Animals And The Effects Of Active Ingredient On CYP2E1

This dissertation is composed of four chapters. Chapter I, serum pharmacochemistry of Rubiacordifolia L．petroleum ether extracts was investigated. The absorption and metabolism in the blood ofrabbit of petroleum ether extract were analyzed rabbit. Chapter II, the metabolism of RMC in rats wasassayed using the method of HPLC. Chapter III, The influence of RMC on mice hepatic microsomalenzymes CYP2E1was investigated by the method of probe drug in vitro. Chapter Ⅳ, the progressofnatural products oncytochrome P450enzymes was summarized.Chapter I. Serum pharmacochemistry of Rubia cordifolia petroleumether extracts.Healthy rabbit was fasted for12hours, and the concentration of petroleum ether extracts was0.2g mL^-1to intragastric administration as dose of5mL·100g^-1. Collect3mL blood before intragastricadministration, and blood was collected at15,30,60,90and120minutes after intragastric administrationrespectively. Serum was pretreated and filtrated with0.45μm micropore filter, and then was analyzed byHPLC. The HPLC system conditions waere Purospher star RP-C18column （4.6mm×250mm,5μm） at25℃, MeOH-H2O-tetrahydropyrane （90:9.3:0.7） mixture as mobile phase, detection wavelength at250nm, flow rate of1.0mL·min^-1. RMC had been absorbed into the blood after15min and had beenmetabolized after30min. After90min, RMC was appaerd that might be transformed by other compoundsin extracts and disappeared after120min.Chapter II. Research metabolism of RMC in rats.Healthy SD rats were fasted for12hours, the1.75mg mL^-1concentration of RMC was injectedin caudal vein acorrding to0.2mL·100g^-1. Collect blood before intragastric administration, and blood wascollected at15,30,60,90and120minutes after injection in respectively. Serum was pretreated and filtrated with0.45μm micropore filter, and then was analyzed by HPLC. The HPLC system consisted ofPurospher star RP-C18column （4.6mm×250mm,5μm） at25℃, MeOH-H2O-tetrahydropyrane （90:9.3:0.7） mixture as mobile phase, detection wavelength at250nm, flow rate of1.0mL·min^-1. RMC had notbeen found in all serum samples. RMC had been absorbed and metabolized in5min.Chapter III. Analyse effects of RMC on mice hepatic microsomalenzymes CYP2E1in vitroHealthy SD rats were put to death after fasted for12hours. The liver was took out and rinsed. Livermicrosome was prepared by method of high speed centrifugation. Protein content was measured withCoomassie light blue. The volume of the incubation system was500μL. The system was made up of80μLLiver microsome,5.0mmol·L^-1MgCl2，0.7mg·mL^-1P-nitrophenol as probe drug and sample10μL. Theexperimental groups was mixed1mg·mL^-1RMC and the blank groups PBS buffer as sample. The systemwas mixed up and incubated for5min in37℃. The reaction was started by1mmol·L－1NADPH. Theexperimental groups and the blank groups were pulled out after30min and60min. The reactions werefinished by mixed up with cold methanol solution. All groups were filtrated with0.45μm micropore filter,and then were analyzed by HPLC. The HPLC system consisted of Purospher star RP-C18column （4.6mm×250mm,5μm） at25℃,1%HAC MeCN-0.05mol L^-1potassium dihydrogen phosphate （20:80） mixtureas mobile phase, detection wavelength at275nm, flow rate of1.0mL·min^-1.4-nitro catechins was foundin both blank groups but not found in both experimental proups. Results showed that RMC is enzymeinhibitors for CYP2E1.Chapter Ⅳ. The progress of natural products on cytochrome P450enzymes.