Résumé
ObjectiveTo investigate the detoxification mechanism of Chebulae Fructus， Glycyrrhizae Radix et Rhizoma and Prepared Aconiti Kusnezoffii Radix Cocta， and their effective components ellagic acid， liquiritin and aconitine based on cardiac cytochrome P450 （CYP450） system. MethodIn in vivo experiments， rats were randomly divided into control group， prepared Aconiti Kusnezoffii Radix Cocta group （0.25 g·kg-1）， Chebulae Fructus group （0.252 g·kg-1）， Glycyrrhizae Radix et Rhizoma group （0.25 g·kg-1） and combination group （0.25 g·kg-1 Chebulae Fructus+0.25 g·kg-1 Glycyrrhizae Radix et Rhizoma+0.25 g·kg-1 prepared Aconiti Kusnezoffii Radix Cocta， with prepared Aconiti Kusnezoffii Radix Cocta as standard）. After 8 days of administration， creatine kinase （CK） and lactate dehydrogenase （LDH） in rats were detected to observe the pathological changes of heart tissue. Real-time PCR and Western blot were performed to detect the mRNA and protein expressions of CYP2J3， respectively. In in vitro experiments， control group， aconitine group， ellagic acid group， liquiritin group and combination group （aconitine+ellagic acid+liquiritin） were set， and their effects on cell number， DNA content， reactive oxygen species （ROS） and mitochondrial membrane potential （MMP） were detected by high content analysis. The changes in the mRNA and protein expressions of CYP2J3 were also observed. ResultIn vivo experiments， compared with the control group， the prepared Aconiti Kusnezoffii Radix Cocta group had increased CK and LDH in serum （P<0.05， P<0.01）， while the combination group had decreased activities of CK and LDH. Additionally， pathological staining results showed that Chebulae Fructus and Glycyrrhizae Radix et Rhizoma reduced the cardiac toxicity caused by prepared Aconiti Kusnezoffii Radix Cocta. Real-time PCR found that compared with the control group， prepared Aconiti Kusnezoffii Radix Cocta down-regulated the mRNA level of CYP2J3 （P<0.05）， while up-regulated that expression when used in combination with Chebulae Fructus and Glycyrrhizae Radix et Rhizoma （P<0.01）. The protein and mRNA translation levels were basically consistent. In vitro experiments， high content analysis revealed that there was a decrease in the cell number， DNA content and MMP fluorescence value of the aconitine group （P<0.01） and the combination group （P<0.05， P<0.01）， and the fluorescence value of the combination group was higher than that of the aconitine group. Moreover， aconitine down-regulated the mRNA level of CYP2J3 （P<0.05）， but the down-regulating ability of aconitine was reversed in the combination group （P<0.05）. ConclusionThe detoxification mechanism of combined Chebulae Fructus， Glycyrrhizae Radix et Rhizoma and prepared Aconiti Kusnezoffii Radix Cocta is mainly that the combination of ellagic acid， liquiritin and aconitine can up-regulate the expression of CYP2J3， and promote the metabolism of arachidonic acid （AA） to produce epoxyeicosatrienoic acids （EETs）， thus reducing the cardiac toxicity， and this effect may start from the transcriptional link.
Résumé
OBJECTIVE: To compare the effect on CYP450 isoenzyme in rats with acute liver injury induced by different chemicals. METHODS: Acute liver injury model of rats induced by tetrachloromethane(CCl4), D-aminogalactose(D-GalN)/lipopolysaccharide(LPS), α-naphthyl isothiocyanate(ANIT) respectively whereas the normal Wistar rats were used as controls. After the tail vein injection with Cocktail probe solutions prepared with five CYP450s probe substrates (phenacetin-CYP1A2, omeprazole-CYP2C9, tolbutamide-CYP2C19, dextromethorphan-CYP2D6, midazolam-CYP3A4), the blood samples were collected from the fundus venous plexus of rat at different time point, the blood drug concentration of the five probe substrates were determined by LC-MS/MS, and the pharmacokinetic parameters were calculated by PK Solutions 2™. Compared with normal rats, the changes of the probe drug pharmacokinetics in different rat models were used as the basis for the evaluation of the metabolic activity of CYP450 isoenzyme. RESULTS: Compared with normal rats, the activities of CYP1A2, CYP2C9, CYP2C19, CYP2D6 of CCl4 group rats were significantly inhibited, and the activities of CYP3A4 was slightly inhibited; the activities of CYP2C9, CYP2D6 and CYP3A4 of D-GalN/LPS group rats were significantly induced, and the activity of CYP2D6 and CYP3A4 was slightly induced, and the activity of CYP1A2 was not significantly affected, but the activity of CYP2C19 was significantly inhibited; the activities of CYP2C9, CYP2C19 and CYP3A4 of ANIT group rats were significantly induced, the activity of CYP3A4 were slightly induced, and the activity of CYP2D6 was not significantly affected, but the activity of CYP1A2 was significantly inhibited. CONCLUSION: There are significant differences in the activities of CYP450 isoenzyme in the rat model of acute liver injury induced by different chemicals.
Résumé
Objective: To investigate and evaluate the effect of components in Biqi Capsulae prescription compatibility on activities of cytochrome P450 (CYP450) in vivo. Methods: A new "Cocktail" one point method has been established, including five probes of Phenacetin, Tolbutamide, mephenytoin, Dextromethorphan, and midazolam. An LC-MS/MS analytical method has been established to determine the above five probes and their corresponding metabolites to analyze and evaluate the potential in vivo induction and (or) inhibition of components from Biqi Capsulae on the above five CYP450 in rats. Results: Biqi Capsulae prescription has compatibility based on CYP450. Compared with blank group, groups of "principal + assistant", "principal + mediator", "principal + assistant + complement", "principal + assistant + mediator", "principal + complement + mediator", and whole Biqi Capsulae could significantly induce the activity of CYP1A2. Moreover, groups of "principal + assistant", "principal + assistant + complement", "principal + assistant + mediator", and whole Biqi Capsulae could significantly induce the activity of CYP2C9. Conclusion: Biqi Capsulae prescription has significant compatibility based on CYP450. These results provide the important information and establish good foundation for further investigation on scientificalness and safety of Biqi Capsulae prescription compatibility.
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Objective To investigate the effect of Honghua Injection on the activity of rat liver CYP2D6.Methods The metabolic rates of probe dextromethorphan(DM) in the blank and Honghua Injection groups(0.9,1.8,and 3.6 mL/kg) in vivo urine and in vitro liver microsome incubated system were determined by high performance liquid chromatography(HPLC).The variation of the metabolic rate of DM represented the effect of Honghua Injection on the activity of rat liver CYP2D6 in vivo and in vitro.Results In vivo and in vitro,the DM metabolic rates of treated groups(1.8 and 3.6 mL/kg) are lower than that in the blank group(P