| Aim:To clarify the mechanism of compatibility and the basis material involved in the treatment of acute pancreatitis in Dahuang Fuzi Decoction.Method:Dahuang Fuzi Decoction,a widely used traditional Chinese medicine(TCM)decoction to treat acute pancreatitis,was originally described in Synopsis of Golden Chamber(JinKui Yao Lue).It is composed of three herbs including Radix et Rhizoma Rhei(Dahuang),Radix Aconiti Lateralis Praeparata(Fuzi)and Radix et Rhizoma Asari(Xixin).Their compatibility ratio is 3:4:1.Metabonomics and pharmacokinetic have been used to clarify the mechanism of compatibility.Endogenous compounds have been analyzed by GC-MS.R2 software has been used to rectify the retention time,peak area and m/z after using Chemstation to export AIA files.Simca-P has been applied to analyze the data using OPLSD calculation.According to the obtained data,the model prediction is satisfied.It has been validated that compounds with VIP value larger than 1.5 and satisfied RSD are possible metabolic markers.Succinic acid,an important intermediate in the citric acid cycle,increased significantly in Dahuang group(p<0.01)compared with control group,indicating that Dahuang inhibit the activity of succinate dehydrogenase.VWhile there is no significant difference between Dahuang Fuzi Decoction and control group.Citric acid cycle happens mainly in mitochondria.Mitochondria damage causes the activity of succinate dehydrogenase irreversible decline,suggesting Dahuang cause the mitochondria damage leading to the nephrotoxicity.The concentration of rhein,emodin and aleo-emodin in plasma have been quantitively analyzed by LC-MS,while physcion and chrysophanol in plasma has been quantified by LC-MS/MS.The method of analysis of rhein,emodin,aloe-emodin,physcion and chrysophanol in vivo has been validated.The biosample extraction method is liquid-liquid extraction.LC-MS has been applied to quantify the concentration of four alkaloids.The method of quantification of benzoylhypaconine,benzoylmesaconine,benzoylaconine and hypaconitine in vivo has been validated.Analytes have been extracted from plasma using liquid-liquid extraction method.Pharmacokinetic parameters have been calculated using WinNonlin software.Comparative pharmacokinetic study of the five anthraquinone from Dahuang and Dahuang Fuzi Decoction and the four alkaloids from Fuzi and Dahuang Fuzi Decoction has been completed.The pharmacokinetic parameters difference including AUC,tmax,Cmax,t1/2 manifested the compatibility mechanism of Dahuang Fuzi Decoction.Peak time and Cmax of anthraquinones prolonged and decreased,indicating combination ease the strong effect of Dahuang.tmax in Dahuang Fuzi Decoction Group prolonged compare with the Fuzi Group,indicating that toxicity compound absorption delayed after compatibility in order to ensure the safety in clinical.PK-PD and spectrum-effect methods have been applied to clarify the basis material of Dahuang Fuzi Decoction.The plasma concentration-time curve has been established.Acute pancreatitis model of rat has been established,inserting a syringe into the bile-pancreatic duct via duodenum papilla in a retrograde way.Another microvascular clamp was used to temporarily nip the duct head.2.5%Sodium taurocholate was transfused into the bile-pancreatic duct at the speed of 0.2 mL/min.Protective ration that represents the PD has been detected at 1 h,3 h,6 h,9 h,12 h,24 h after the surgery.The effect-time curve has been established.PK-PD correlation has been calculated using MATLAB software.Levenberg-Marquardt algorithm calculated the value of r and l.The result showed that chrysophanol and aloe-emodin are the basis materials involved in the treatment of acute pancreatitis of Dahuang Fuzi Decoction.Compounds from different compatibility group have been analyzed using HPLC-Q-TOF-MS.Peakview(?)software has been applied to find the target compounds and calculated the peak area.Spectrum-effect has been calculated by SPSS 16.0 using Pearson algorithm method.14 compounds have been found to be the basis materials involved in the treatment of acute pancreatitis of Dahuang Fuzi Decoction,including rhein,physcion,emodin,benzoylmesaconine,neoline,talatisamine and so on.Result:Above method have been applied successfully to clarify the mechanism of compatibility and the basis material involved in the treatment of acute pancreatitis in Dahuang Fuzi Decoction... |
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