ABSTRACT
The study was designed to explore the effects of HS060098 on activation of peroxisome proliferator-activated receptors (PPARα, γ and δ) and in the down-regulation of hyperlipidemia in golden hamster. Luciferase gene reporters of PPARα, PPARγ and PPARδ were constructed in HepG2 cells and the green fluorescent protein (GFP) was used as an internal reference. Transfected cells were then cultured with various concentrations of HS060098 for 24 h. The peroxisome proliferator-response element luciferase activity was determined by the dual-luciferase reporter gene assay system. To investigate the lipid-lowering effect of HS060098, hyperlipidemic golden hamsters fed by high-diet were administered orally with HS060098 through prophylactic and therapeutic approaches respectively. The levels of blood lipids such as total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) and fat index in hamsters were evaluated. The results showed that HS060098 was a potent activator of PPAR δ with a good selectivity and the median effective concentration (EC 50) is 0.01 μmol·L-1, while no obvious PPARα and PPARγ activation was observed. In the golden hamster, oral administration of HS060098 (5, 10, 20 mg·kg-1·d-1) for 2 weeks, led to a significant decrease the concentrations of plasma TC, TG, LDL-C and fat index (PPPPδ agonist with a significant activity in the prevention and therapy of hyperlipemia in golden hamster.
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
To study the pharmacokinetic process of Danshensu in cerebal ischemia injury model rats and the correlation with its anti-cerebral ischemia effect. In this study, the middle cerebral artery occlusion (MCAO) model was established, in which all of the rats were intravenously injected of Danshensu at a single dose of 40 mg x kg(-1). The HPLC-DAD method was applied to determine the plasma concentration of Danshensu at different time points and draw the drug-time curve. Meanwhile, the superoxide dismutase (SOD) and the lactate dehydrogenase (LDH) activity were determined to draw the time-effect curve. The DAS 3.2. 6 software was used to process the data, analyze their correlation, compare the pharmacokinetic difference between model and normal rats after the administration of the same doses of Danshensu and the changes in pharmacodynamic indicators of model rats after the administration, and evaluate the effect of Danshensu in treating the cerebral ischemia disease. According to the results, the pharmacokinetic processes of Danshensu in the cerebral ischemia-reperfusion and normal rats were consistent to the two-compartment model. The main pharmacokinetic parameters were: t1/2alpha were (0.267 +/- 0.026), (0.148 +/- 0.020) h;t1/2beta were (1.226 +/- 0.032), (1.182 +/- 0.082) h; AUC0-infinity were (42.168 +/- 4.007), (26.881 +/- 1.625) mg x L(-1) x h. After the cerebral ischemia-reperfusion, the activity of SOD decreased and the activity of LDH increased. Danshensu could inhibit the decrease in the SOD activity and the increase in the LDH activity within a certain period of time. This indicated that Danshensu could stay longer in cerebral ischemia-reperfusion rats than in normal rats and eliminated more slowly, which reflected the rationality of Danshensu in the clinical treatment of cerebral ischemia diseases. Danshensu's effect against the cerebral ischemic injury may be related with its level in vivo. Its plasma concentration is positively related to the SOD activity and negatively related to the LDH activity.