Effects Of Zhenwu Decoction On Activity And MRNA Expression Of Cytochrome P450 Enzymes In Normal And Chronic Heart Failure Rats

Chronic heart failure is the end stage of the most heart diseases,and the purpose of therapeutic is to improve the quality of life of patients in clinically.At present,the treatment of chronic heart failure is mainly based on western medicine,but there are many side effects and the treatment can only change the quality of life.Chinese compound formula has the characteristics of multi-component,multi-target,multi-effect integration,with few side effects.Clinical experience of Zhenwu Decoction,a representative of Chinese compound formula,shows that Zhenwu Decoction has significant curative effect on chronic heart failure.Traditional Chinese medicine has two-way regulation effect on individuals in different states.The role and function of Chinese compound formula in normal state and disease state may be different.Therefore,the regulation of P450 enzyme expression and activity may be completely different,in view of the clinical treatment of Zhenwu Decoction.Chronic heart failure often requires a combination of drugs,so the study of the effect of Zhenwu Decoction on the expression and activity of P450 enzyme in rats under normal rat and chronic heart failure disease has important clinical significance.Objective: This study was to investigate the effect of Zhenwu Decoction(ZWD)on the activity and m RNA expression of seven hepatic cytochrome P450(CYP450)isozymes in normal and chronic heart failure(CHF)rats.It provides theoretical guidance in the safe use of clinical medicines for patients with CHF,and helps to promote the further development of ZWD in the treatment of CHF.Methods:(1)The UPLC-MS/MS method was established to simultaneously determine the corresponding probe substrates of seven major CYP450 enzymes(CYP1A2,CYP2B1,CYP2C6,CYP2C7,CYP2C11,CYP2D2,CYP3A1)in rats(phenacetin,bupropion,diclofenac,amodiaquine,omeprazole,dextromethorphan,midazolam);(2)54 SD animals were divided into normal state group(n=24)and CHF state group(n=30).The normal state group of experimental animals were normally raised and divided into 4 groups: normal saline group,ZWD low,medium,high(2.1875,4.375,8.75g/kg)dose group;Meanwhile,the experimental animals in CHF disease state group were replicated in a rat model of CHF by coronary artery ligation,and were divided into 5 groups: sham operation group,model group,ZWD low,medium,high(2.1875,4.375,8.75g/kg)dose group.All of experimental animals were continuously intragastric administrated for 4 weeks,and after the last day of administration,they were injected with probe drug mixture solution in the tail vein respectively,then the plasma samples were collected from the fundus at the preset time point.Seven probe substrates concentration in plasma of different groups were determined by UPLC-MS/MS method,and calculated the main pharmacokinetic parameters by DAS 2.0.Statistical analysis was performed to evaluate the effect of ZWD on CYP450 enzyme activity in rats of all groups;Additionally,detecting the effects of ZWD on the changes of hemodynamic parameters in CHF rats,evaluating the relationship between CYP450 enzyme activity and CHF by analyzing the correlation coefficients between the main pharmacokinetic parameters and the main hemodynamic parameters.(3)At the same time,27 male rats of the same batch of SD were divided into normal state group(n=12)and CHF disease state group(n=15),the modeling and grouping methods were the same as the above study of CYP450 enzyme activity.Analogously,after continuous administration for 4 weeks,liver tissues were taken to determine the m RNA expression of CYP450 isoforms by RT-q PCR technology in each group.Results:(1)A simple and efficient UPLC-MS/MS method was applied to determinate phenacetin,bupropion,diclofenac,amodiaquine,omeprazole,dextromethorphan,midazolam simultaneously.(2)In normal rats,ZWD can induce CYP2C6,CYP2C11 and CYP3A1 enzyme activities,but had no effects on CYP1A2,CYP2B1,CYP2C7 and CYP2D2 enzyme activities;CYP1A2,CYP2B1,CYP2C7,CYP2C11 and CYP3A1 enzyme activities in CHF disease state rats were significantly reduced,and the low,medium and high doses of ZWD can reverse the changes of CYP450 enzyme activities in CHF disease state.At the same time,there was a significant positive correlation between phenacetin,diclofenac,omeprazole and midazolam AUC and LVEDP(P < 0.05).(3)In normal rats,low,medium and high doses of ZWD can regulate the expression levels of CYP2C6,CYP2C11 and CYP3A1 m RNA at different degrees,and had no effect on the expression levels of CYP1A2,CYP2B1,CYP2C7 and CYP2D2 m RNA;The expression levels of CYP1A2,CYP2B1,CYP2C6,CYP2C11 and CYP3A1 m RNA were significantly decreased in CHF disease state rats,and the ZWD could reverse the expression of CYP450 isoform m RNA in CHF disease.The m RNA expression of CYP1A2,CYP2C6,CYP2C11 and CYP3A1 was significantly negatively correlated with the main hemodynamic parameter LVEDP(P < 0.05).Conclusion: In the CHF disease state,the activity and m RNA expression of five CYP450 enzymes changed in rats.ZWD can reverse these changes and lead some of the four CYP450 enzymes to be normal,which suggests that the changes of CYP450 enzymes have instructive meaning in CHF disease.Additionally,when ZWD used together with other medicines,the potential interactions should be taken into consideration for the treatment of CHF.