ABSTRACT
Objective: Using total quantum statistical moment analysis to evaluate the influence of the main effective constituents in Danhong Injection on the total pharmacokinetic parameters after orthogonal design to research internal compatibility rule of traditional Chinese medicine (TCM) compound. Methods: Using modified middle cerebral artery occlusion (MCAO) method to establish rat disease model of focal cerebral ischemia-reperfusion (I/R) injury. According to the orthogonal design L9 (34) composed of the main effective constituents (danshensu, protocatechuic aldehyde, salvianolic acid B, and HYSA) of Danhong Injection, nine combinations with different dosage ratios were formed, which were used for iv administration in MCAO rats to study the pharmacokinetics of effective constituents after compatibility. Using non-compartmental model of DAS 3.2.6 software to fit the pharmacokinetic parameters and using total quantum statistical moment analysis method to calculate the main total quantum statistical moment parameters. The influence of different compatibilities on the total pharmacokinetics parameters was analyzed by orthogonal test. Results: There were different effects of the main effective constituents with orthogonal compatibilities on the total quantum statistical moment parameters. Danshensu had the strongest effect on the total AUC and had a significant effect (P < 0.05). A3B3C2D1 was the preferred dose combination for AUCt. HYSA had the strongest effect on the total MRT and its optimal ingredient composition was A1B3C1D1. Conclusion: The total quantum statistical moment analysis could combine the pharmacokinetics of danshensu, protocatechuic acid, vanillic acid, salvianolic acid B, and HYSA, and express the total pharmacokinetic behaviors. The total quantum statistical moment analysis method can be used to study the pharmacokinetics of multi-component in TCM compound prescription, which can provide the method for research on internal compatibility rule for TCM compound.
ABSTRACT
Objective: To develop an HPLC-DAD method for the simultaneous determination of danshensu, protocatechuic acid, vanillic acid, salvianolic acid B, and hydroxysafflor yellow A (HYSA) in cerebral ischemic injury of rats, in order to provide an accurate and reliable analytical method to study the pharmacokinetics. Methods: The middle cerebral artery occlusion (MCAO) model was established, in which all the rats were iv injected with the active constituents in Danhong Injection simultaneously (danshensu, protocatechuic aldehyde, salvianolic acid B, and HYSA). The analysis of the measured components' plasma concentration was carried at 30℃ on the reversed-phase column of Agilent Eclipse XDB-C18 (150 mm×4.6 mm, 5 μm) and eluted with acetonitrile and water containing 0.4% phosphate acid as mobile phase in gradient mode. Detection wavelengths were 280 nm for danshensu, protocatechuic acid, vanillic acid, salvianolic acid B and 403 nm for HYSA. The propyl p-hydroxybenzoate was used as the internal standard (IS). Results: Protocatechuic aldehyde rapidly metabolized in MCAO rats, but its metabolites, protocatechuic acid and vanillic acid, were easily detected. The linear ranges of danshensu, protocatechuic acid, vanillic acid, salvianolic acid B, and HYSA were 0.35-140 (R2=0.9998), 0.15-60 (R2=0.9990), 0.05-20 (R2=0.9988), 0.25-100 (R2=0.9996), and 0.075-30 mg/L (R2=0.9985), respectively. The mean recoveries were between 80% and 120% and the RSD value of intra-day and inter-day was less than 10%. The results of stability meet the requirements for biopharmaceutical analysis. The main pharmacokinetic parameters of danshensu, protocatechuic acid, vanillic acid, salvianolic acid B and HYSA were as follows: AUC0~∞ were (1143.862±230.840), (427.024±59.293), (135.785±47.631), (418.631±66.242), (288.788±87.809) mg·min/L and t1/2z were (71.496±29.067), (82.379±26.279), (40.331±6.006), (125.164±59.709), (177.577±112.836) min. Conclusion: The method is simple, rapid, precise and stable, which can be used to the pharmacokinetic studies on the compatibilities of danshensu, protocatechuic aldehyde, salvianolic acid B, and HYSA in MCAO rats.
ABSTRACT
Aim This was to study the effects of hydroxysafflor yellow A(HYSA)on iNOS expression in rats following focal cerebral ischemia. Methods Rat cerebral ischemia-reperfusion injury was induced by middle cerebral artery occlusion. The expression levels of iNOS were examined using immunohistochemical method. The neurological outcome and the cerebral infarct area were evaluated. Results The expression levels of iNOS in HYSA groups were significantly lower than those in ischemic model group. Treatment with HYSA also decreased the cerebral infarct area and the neurological deficit score. Conclusion HYSA induced down regulation of iNOS expression, which may mediate the protective effect of HYSA on cerebral ischemia.