Integrated Pharmacokinetic And Pharmacodynamic Study Of Fuzi On 'Yang Deficiency'

Object:To study the material basis and the mechanism of effect of'wenyang'of fuzi on treatment of two kinds of classical'yang deficiency'model,including constipation and heart disease with'yang deficiency'by the pharmacokinetic and pharmacodynamic methods.Total alkaloids from fuzi are the main active compound in fuzi.It has the function of'wenyang'and purgation.Therefore,we used total alkaloids from fuzi for the treatment of constipation rats with'yang deficiency'by the method of integrated pharmacokinetic and pharmacodynamic study.However,fat-soluble alkaloids in total alkaloids from fuzi are toxic,while water-soluble alkaloids have'wenyang'and cardiotonic effects.Therefore,we selected water-soluble alkaloids for the integrated pharmacokinetic and pharmacodynamic study of chronic heart failure in rats.Method:In the pharmacokinetic and pharmacodynamic experiment of total alkaloids from fuzi in constipation rat with yang deficiency,the rats were randomly divided into normal rat group and constipation rat group with'yang deficiency'.And they were divided equally into low,medium,and high dose group,respectively.A single dose of9.6 mg/kg,19.2 mg/kg,and 38.4 mg/kg total alkaloids from fuzi were orally administered to the three dose groups,respectively.A series of blood samples were collected through orbit venae angularis of unanaesthetized rats by nicking method before and after oral administration at time points of 0 min,5 min,10 min,20 min,30min,40 min,60 min,120 min,180 min,300 min,420 min,540 min,720 min,1440min into heparinized tubes.And the plasma samples were extracted by liquid–liquid extraction.Then,an ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry was used to detect the plasma concentration of aconitine,mesaconitine,hypaconitine,benzoylaconine,benzoylmesaconine,benzoylhypaconitine,and aconine of total alkaloids from fuzi in different time points.And the pharmacokinetic process was studied.The contents of motilin,gastrin,endothelin and vasoactive intestinal peptide were measured by multiscan spectrum.Finally,principal component analysis was used to analyze the plasma concentration and hormone content in order to elucidate the material basis and mechanism of fuzi in the treatment of constipation rats with'yang deficiency'.In the pharmacokinetic and pharmacodynamic experiment of fuzi water-soluble alkaloids in chronic heart failure rats,the low,medium,high dose group of normal and chronic heart failure rats were respectively given 0.072 g/kg?0.143 g/kg?0.286g/kg fuzi water-soluble alkaloids with once a day for a week.A series of blood samples were collected through orbit venae angularis of unanaesthetized rats by nicking method before and after oral administration at time points of 0 min,3 min,5min,10 min,20 min,40 min,60 min,90 min,120 min,240 min,360 min,540 min,720 min,1440 min into heparinized tubes.And a protein precipitation method was taken to treat plasma samples.Then,an ultra performance liquid chromatography was used to detect the plasma concentration of 3-hydroxytyramine hydrochloride,uracil,salsolinol,uridine,3-methoxy-p-tyramine hydrochloride,guanosine,and higenamine in fuzi water-soluble alkaloids in different time points.The mean drug concentration-time curve was plotted and the pharmacokinetic parameters were calculated.The contents of angiotensin?,aldosterone,atrial natriuretic peptide,endothelin,and brain natriuretic peptide were determined by multiscan spectrum.Finally,principal component analysis was used to analyze the plasma concentration and the content of hormone in order to elucidate the material basis and mechanism of fuzi water-soluble alkaloids in the treatment of chronic heart failure rats.Results:C_max and AUC_last of aconitine,mesaconitine,hypaconitine,benzoylaconine,benzoylmesaconine,benzoylhypaconitine,and aconine were increased with increasing dosage in the range from low dose to high dose.The in vivo process of total alkaloids from fuzi was basically in line with linear kinetic characteristics.Total alkaloids from fuzi could increase plasma concentration of motilin,gastrin,endothelin and vasoactive intestinal peptide.The integrated pharmacokinetic and pharmacodynamic data of total alkaloids from fuzi showed obvious pharmacokinetic process and pharmacodynamic characteristics,and the pharmacokinetic and pharmacodynamic characteristics of total alkaloids from fuzi after integration were basically consistent with those before.It could express the pharmacokinetic and pharmacodynamic characteristics of total alkaloids from fuzi.In the range of dosages,C_max and AUC_last of 3-hydroxytyramine hydrochloride,uracil,salsolinol,uridine,guanosine,and higenamine were increased with increasing dosage,the in vivo process of fuzi water-soluble alkaloids was basically in line with the linear kinetic characteristics in nomal rats.After administration of fuzi water-soluble alkaloids,the level of angiotensin?,aldosterone,atrial natriuretic peptide,endothelin and brain natriuretic peptide was decreased in heart failure rats.The integrated pharmacokinetic and pharmacodynamic data of fuzi water-soluble alkaloids showed obvious pharmacokinetic process and pharmacodynamic characteristics,and the pharmacokinetic and pharmacodynamic characteristics of fuzi water-soluble alkaloids after integration were basically consistent with those before.It could express the pharmacokinetic and pharmacodynamic characteristics of fuzi water-soluble alkaloids.Conclusion:The results showed that total alkaloids from fuzi were the main material basis of fuzi in the treatment of constipation rat with'yang deficiency'.At the same time,fuzi water-soluble alkaloids were the main material basis of fuzi in the treatment of chronic heart failure in rats.Total alkaloids from fuzi mainly promote the secretion of motilin,gastrin,endothelin,and vasoactive intestinal peptide to achieve effect of'wenyang'.Fuzi water-soluble alkaloids mainly suppress and decrease the secretion of angiotensin?,aldosterone,atrial natriuretic peptide,endothelin and brain natriuretic peptide to achieve effect of'wenyang'.The purpose of this study is to reveal the material basis and mechanism of effect of'wenyang'of fuzi,and provide theoretical basis for rational use of fuzi in clinic.