| Objective: Vascular endothelial cells are the barrier between blood and organism. As an active metabolic pool, it can secrete lots of vasoactive substances. So vascular endothelial cells (VEC) are very important to vasomotor function and hemokinesis. In order to search the medicine that can protect the VEC, we research the protective effect on VEC of the injection of radix salviae miltiorrhizae (the water-soluble preparation of radix salviae miltiorrhizae) and tanshinone (the lipid soluble element of radix salviae miltiorrhizae). Because the injection of radix salviae miltiorrhizae is widely used, we also research the pharmacokinetics of danshensu in order to offer the theoretical base for clinical medication. Method: Experiment 1. 120 healthy Wistar rats were divided randomly into 8 groups:Normal group, model of blood stasis group, the injection of radix salviae miltiorrhizae group [high dose (4ml/kg), middle dose (0.4ml/kg), low dose (0.08ml/kg)] ,tanshinone group [high dose (90mg/kg), low dose (10mg/kg) ]and nifedipine (2.5mg/kg) group. Each rat was administered continuously for 5 days. On day 4, 30 minutes after administration, the blood stasis model was established with adrenaline and ice stimulation. On day 5, 30 minutes after administered, we collected the blood of rats to determine the blood viscosity, hematocrit, the concentration of fibrinogen, circulating endothelial cell (CEC)count, the concentration of NO in plasma and the concentration of endothelin (ET) in plasma.Experiment 2. 8 healthy rabbits were intramuscularly injected the injection of radix salviae miltiorrhizae with 2ml/kg. The blood of rabbits was collected at different time before and after administration. 1 mol/L HC1 was used to precipitate protein; acetic ether was used to extract the danshensu; After blowed by N2 at 37 C, the extract was determined by HPLC method. In this system, The column was C18 (5um,250 X 4.6 mm);The mobile phase consisted of methanol and 1.75% glacial acetic acid solution(l :9), Flow rate was 1.0ml/min; The UV detector was set at 280nm; P-hydroxybenzoic acid was used as the internal standard. According tothe result, we drawed the concentration-time curve and calculated the pharmacokinetic parameters of danshensu.Results: Experiment 1. In blood stasis rats, the blood viscosity and the concentration of fibrinogen were elevated (P<0.01) , hematocrit trended to increase. The VEC of the blood stasis rats was damaged: Compared with the normal group, CEC increased (P<0.01) , the concentration of NO in plasma was lower (P<0.05) , the concentration of ET increased (P<0.01) .The injection of radix salviae miltiorrhizae can improve the hemorheology:in the middle and high dose groups, the blood viscosity and the concentration of fibrinogen were cut down (P<0.01) .The injection of radix salviae miltiorrhizae had protective effect on VEC: to decrease CEC; to elevate the concentration of NO,the effect was similar to that of nifedipine. But the effect on ET was not shown(P > 0.05).Though it had no effect on hemorheology, tanshinone had a better effect on VEC . Same as that of nifedipine , tanshinone can protect VEC by decreasing CEC (P<0.01) and elevating the concentration of NO (P<0.01) . Tanshinone can antagonize the effect of ET. In the low and high dose groups, the concentration of ET was degraded conspicuously (P<0.01) .Experiment 2. The established HPLC method for the determination of danshensu is excellent in specificity, precision and accuracy. After intramuscularly injected the injection of radix salviae miltiorrhizae, danshensu is absorbed and eliminated fast in vivo. Tpeak is 19 minutes, t1/2 is 81 minutes. Metabolism of danshensu is fitted to first-rate one-compartment model.Conclusion: Blood stasis is correlated with the damage of the VEC. The injection of radix salviae miltiorrhizae and tanshinone have protective effect on VEC. The mechanism of this action is to degrade shear stress of blood stream by improving blood hemorheology. Tanshinone has a better effect on decreasing ET , so that we suggest it m... |
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