| Objective:This subject was based on the effective active ingredient Sal B and paeonol from traditional medicine Chinese Shuangdan compound, designed by changes in the baseline geometric, and the effects of different ratio combinations on relaxation of isolated rat arteries and myocardial hypoxia/reoxygenation injury protection was observed, and also compared the effects of different combinations on acute myocardial ischemia-reperfusion injury (MI/RI) protection, ultimately, selected the Sal B/Pae best ratio combination; also compared the Sal B/Pae combinations on endothelial cells P-selectin expression in MI/RI by Western blot, so as to provide some methods and ideas for a new and relatively clear mechanism and indications Chinese herbal formula.Methods:The effect of different Sal B/Pae combinations protection on MI/RI was compared with the different levels of organs, cells, molecular and animals. Observed the different combinations on relaxation of isolated rat arteries, and then the different Sal B/Pae combinations on primary cardiomyocyte hypoxia/reoxygenation injury protection was also compared, primary cardiomyocyte wasn't cultured at least 36h until beating blance, and then hypoxia lh reoxygenation 3h to simulated ischemia and reperfusion injury, add in combinations 3hs before hypoxic, observed cell viability, apoptosis and expression of intracellular MDA. In MI/RI, continuous ip administration three days, ligation of the left anterior descending coronary artery, ischemia 30min and reperfusion 120min, then the effect of different Sal B/Pae combinations protection on MI/RI were assessed by myocardial infarction, ECG∑-ST, MDA, NO, SOD, CK-MB and morphology etc. In molecular level, the adhesion molecules of P-selectin in initial injury was observed primarily by Western blot on MI/RI, to explore the Sal B/Pae on the MI/RI of the possible protective mechanism.Results:1. Relaxation of isolated rat arteries in vitro experiments:the effect of different Sal B /Pae combinations on the relaxation of isolated rat arteries was obvious, compared with Sal B/Pae 4/0 and 0/4 group,3/1 and 2/2 of Sal B/Pae combinations have significant vasodilation, in which 3.0/1.0 was the best.2. Primary myocardial hypoxia/reoxygenation injury protection:Sal B, Pae and Sal B /Pae (3/1) at a concentration of 1 mg·mL-1,100μg·mL-1,10μg·mL-1,1μg·mL-1 were observed to cell apoptosis and apoptosis, the Sal B/Pae (3/1) in 10μg/mL concentration has the best effect on cell viability, cell apoptosis rate and the expression levels of intracellular MDA;and Sal B/Pae (3/1) concentration can also reduce intracellular MDA in 100μg·mL-1,10μg·mL-1.3. Acute myocardial ischemia-reperfusion injury protection:the infarction size of other combination groups was more tiny compared with the Model group, and the infarction size of Sal B/Pae 3/1 group and 1/1 group was also smaller than Sal B and Pae unilateral groups, in which Sal B/Pae 3/1 group has the best effect, it can reduce ECG∑-ST, MDA and CK-MB, and increase NO and SOD expression.4. Compared with Model group, the Western blot results show that the concentration of Sal B/Pae 3:1 could reduce the P-selection expression, and the Model group was significantly higher than other groups.Conclusion:1. the Sal B/Pae combinations all have some effects on the relaxation of isolated rat arteries and Primary myocardial hypoxia/reoxygenation injury protection, and the combination groups were more obvious than the unilateral groups.2. The Sal B and Pae combinations have a apparent protection on acute MI/RI, and the best combination was Sal B/Pae 3/1 group; there may exist some synergistic or additive effect between Sal B and Pae in MI/RI protection.3. Sal B and Pae combinations can regulate the expression of endothelial cell P-selectin in MI/RI, This show that Sal B and Pae combinations maybe have some association with the expression of endothelial cell P-selectin on MI/RI protection. |
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