| Objective:Xingnaojing is a flavor-reducing preparation for Angong Niuhuang Wan,which is made of musk,borneol,gardenia and turcuma.It plays an important role in the treatment of cerebral ischemia disease.In this study,YuJin,the main component of Xingnaojing,was selected as the research object.Gas chromatography-mass spectrometry was used to detect and calculate the concentration and pharmacokinetic data of the most important pharmacological active component of curdione,the main component of Xingnaojing,in plasma and brain tissue of rats with focal cerebral ischemia.To investigate the distribution and metabolism of the pharmacological active ingredient of Xingnaojing curdione,in blood and brain tissue after passing the blood-brain barrier,and to provide relevant experimental basis for the follow-up search for pharmacodynamic substances of Xingnaojing playing the role of brain protection.Methods:First,the focal cerebral ischemia model of rats was prepared by blocking the middle cerebral artery with thread embolus method.On the third day after modeling,60 rats were successfully constructed by selecting Longa score 1-3 and divided into two dosage groups A and B,with 30 rats in each group.Group A and Group B were given two kinds of self-made Xingnaojing gavage solution by intragastric administration.The intragastric administration dose of SD rats in group A was 3.6g/kg body weight(equivalent to 0.45g/ml of Xingnaojing gavage liquid in 8ml/kg),and that of SD rats in group B was 7.2g/kg(equivalent to 0.9g/ml of Xingnaojing gavage liquid in 8ml/kg).At 0.5h,1h,1.5h,2h,2.5h,3h,5h,7h,9h,and 12h after administration,3ml of blood was collected from the hearts of 3 rats,and added to the heparinized centrifuge tube.The centrifuge was carried out at 3000 RPM for 10min,and the upper plasma was taken and stored at-20℃ away from light for later use.At the same time,the cranial cavity of the rats was cut out and the brain tissue was immediately stored in liquid nitrogen.To establish a method for the determination of curcuma dione in serum samples and brain homogenate samples by gas chromatography-mass spectrometry.The detected blood/brain drug concentration-time data were fitted and analyzed by DAS2.0 pharmacokinetic software,and the corresponding pharmacokinetic parameters were calculated.Results:1.The establishment of a method for the determination of curdione in Xingnaojing in the plasma and brain tissue of ischemic stroke rats:A method was established for the determination of curdione in the blood and cerebrospinal fluid of ischemic stroke model rats after Xingnaojing intragastric administration:Plasma samples were extracted by methanol protein precipitation and brain tissue samples were extracted by three methanol protein precipitation and then directly injected into gas chromatography-mass spectrometry(GS-MS).Chromatographic mass spectrometry conditions:AgilentDB5MSUltraInert(30m×250μm×0.25μm,);Programmed temperature rise:the initial column temperature was 80℃,held for 2min,at 10℃/min,heated up to 200℃,held for 4min,and increased to 280℃ at 30℃/min.The carrier gas was helium,the flow rate was 1.0ml/min,and the shunt ratio was 10:1.EI ion source Agilent7000C triple quadrupole mass spectrometer,single ion scanning mode,ion source temperature=280℃,electron energy=70eV.The standard curve of curdione of blood samples was y=167.37x-6799.5(=0.996),and the linear range was 97.7-6250μg/mL.The standard curve of curdione in brain tissue was y=274.52-3556.9(=0.993),and the linear range was 39.15000μg/mL.In the linear equation,y is the response value and x is the concentration of curcuma dione.All the above methods have been methodologically verified.2.Pharmacokinetic study of curdione in middle cerebral artery embolization(MCAO)rats after Xingnaojing administration:The measured blood and brain tissue concentration data of the A and B dose groups were processed by DAS2.0 to obtain the pharmacokinetic data:(1)Main pharmacokinetic data of the A dose group:The plasma Cmax was 2185.75±9.61μg/L,AUC(0-t)was 8560.161±62.769μg/L·h,and t1/2β was 5.726±0.508h.Cmax was 1429.513±29.032μg/L,AUC(0-t)was 158.296±7.105mg/L and t1/2β was 8.632±0.327h.Te was 67.41%;(2)The main pharmacokinetic data in B dose group were:plasma Cmax was 2951.44±40.547μg/L,plasma AUC(0-t)was 11668.515±71.603 μg/L·h,plasma t1/2β was 7.2 83±0.405h;Cmax was 2560.14±24.361μg/L,AUC(0-t)10382.497±104.878mg/L,t1/2β8.082±0.135h.Te was 67.41%.Conclusions:1.The cerebral concentration of curdione in Xingnaojing rats in MCAO rats can pass the blood-brain barrier and change with the blood concentration,and all reach the maximum concentration in about 2 hours,but the maximum cerebral concentration is obviously less than the maximum blood concentration(P<0.05).2.The cerebral blood drug ratio in the B dose group was significantly higher than that in the A dose group(P<0.05),and the accumulation of curdione in the brain was more significant after high dose of Xingnaojing administration.3.At different doses of Xingnaojing,the half life of curdione in brain tissue was significantly longer than that in plasma(P<0.05),and the elimination process of curdione in brain tissue was slower than that in plasma after administration of Xingnaojing.4.There was no significant difference in the half-life of curdione in brain tissue between the two groups given different doses of Xingnaojing(P>0.05),and the change of dose of Xingnaojing did not affect the elimination process of curdione in brain tissue.5.The plasma half-life of curcuma dione in B dose group was longer than that in A dose group(P<0.05),and the elimination process of curdione in plasma was affected by the change of Xingnaojing dose. |
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