| Object: To study the effect of Naoluoxintong capsul(NLXTC) on Hemorheology in rats model of cerebral ischemia-reperfusion injury,exploring improving effect on concentration, viscosity, agglutination and coagulability of blood in rats model after cerebral ischemia-reperfusion injury, in order to provide a pharmacological basis for the clinical use of NLXTC.Methods: 1. The establishment and evaluation on rat model of cerebral ischemia reperfusion injury(1)the rats model of middle cerebral artery occlusion-reperfusion(MCAO / R) was established with the suture-occluded method,Longa's score method was used to measure neurological dysfunction in rats at the points of 24 h ? 8d and 14 d after cerebral ischemia-reperfusion injury in rats.(2)The brain water content(BWC) and the brain index was calculated by dry-wet method.(3)Observing the influence of NLXTC on pathological changes of rat brain tissue after cerebral ischemia reperfusion injury by HE staining.2.Effects of NLXTC on blood viscosity and erythrocyte function in rats with cerebral ischemia reperfusion injury using automatic blood rheometer to observe the influence of NLXTC on the plasma viscosity?whole blood viscosity(1s-1,5s-1,50s-1,200s-1)?erythrocyte rigidity index?rythrocyte aggregation index ?hematocrit ?erythrocyte deformation index.3.Effects of NLXTC on coagulation system in rats with cerebral ischemia reperfusion injury ELISA method was used to detect the content of FIB in plasma,Detecting APTT?PT by semi automatic blood coagulation analyzer according to the kit instructions.4.Effects of NLXTC on blood platelet function and thrombosis in rats with cerebral ischemia reperfusion injury Through Born's turbidimetric method by platelet aggregation analyzer for determination of platelet aggregation rate in 1min?2min?5min and the maximum aggregation rate,calculating the inhibition rate of platelet aggregation,to observe its effect on platelet aggregation function in rats model induced by cerebral ischemia-reperfusion injury;According to the kit introduction with enzyme linked immunosorbent assay(ELISA)testing the content of cAMP and vWF in plasma, with enzyme linked immunosorbent assay(ELISA);the method of flow cytometric was used to test CD41/CD61,CD62 P,to explore the effects of NLXTC on the platelet function and thrombosis in rats with cerebral ischemia reperfusion injury.Results:1. The rats model of middle cerebral artery occlusion-reperfusion was successfully produced.(1)NLXTC reduced the neurological scores of rats model after cerebral ischemia reperfusion injury.Longa's scores was used to measure neurological scores at the points of 24 hours ?8days ? 14 days in rats model after cerebral ischemia reperfusion injury.The results showed that the neurological scores was above 2 at the point of 24 hour after operation,compared with the model group, the neurological behavior scores of the sham operation group were significantly increased(P<0.01).the scores of the rats in the model group decreased with time without treatment, which shows the neural function itself has a trend of natural recovery, the neurological score decreased after the treatment of the NLXTC(1.13g/kg?2.25g/kg?4.50g/kg),compared with the model group,the difference was significant in 14 d,which indicated that the recovery of neurological function can be accelerated by NLXTC, with a dosage-dependent trend.(2) NLXTC reduced the brain water content(BWC) and the brain index of rats model after cerebral ischemia reperfusion injury.Compared with the sham operation group,the brain water content(BWC) and the brain index of rats model was increased significantly,which has a significant difference.compared with model group,NLXTC(high?mid and low)dose group can reduce the brain water content(BWC)and the brain index.(3)HE staining results show that obvious brain swelling symptom was found in rats model,compared with the sham operation group,NLXTC can reduce the symptoms.2. NLXTC can improve blood viscosity and red blood cell function in rats with cerebral ischemia reperfusion injury.The whole blood viscosity( 1s-1,5s-1,50s-1,200s-1)and plasma viscosity can be reduced,and the red cell parameters can be improved by NLXTC( high ? mid and low)dose group,erythrocyte rigidity index, erythrocyte aggregation index?hematocrit was decreased,and erythrocyte deformation index was increased,which has statistical significance.3. NLXTC can inhibit the coagulation pathway in rats with cerebral ischemia reperfusion injury.Compared with the sham operation group,the content of FIB was increased in model group, PT, APTT was significantly reduced with a statistical difference, NLXTC prolonged PT,APTT,reduced the content of FIB clearly,which has a statistical difference.4.Exploring the influence and mechanism on platelet function and thrombosis of NLXTC in rats with cerebral ischemia reperfusion injury.4.1 NLXTC prolonged the time of thrombosis by electrical stimulation of carotid artery in rats model induced by the cerebral ischemia reperfusion.Compared with the rats in the sham operation group,the time of thrombus formation of the ones in model group was shortened obviously(P < 0.05),NLXTC each group can prolong the time of thrombusformation.the difference was significant,which indicated that NLXTC has the efffect of anti-thrombus. Compared with the sham operation group, the platelet aggregation rate in 5 minutes was increased, and it has statistical significance, which indicates that it was easier for platelet to aggregate; NLXTC(high?mid?low)dose group has a clear effect on inhibit platelet aggregation, which has statistical significance.4.2 NLXTC can reduce the content of vWF and increasing the content of cAMP in plasma of rats model after cerebral ischemia reperfusion injury.Compared with the sham operation group,the vWF content in plasma of rats model was increased, and the content of cAMP in plasma was reduced(P<0.01),NLXTC can reduce the content of vWF and increasing the content of cAMP in plasma of rats model after cerebral ischemia reperfusion injury, which has statistical significance.4.3 NLXTC can reduce the positive expression rate of platelet membrane glycoprotein CD41/CD61 and mean fluorescence intensity of CD62 p.Conclusion:NLXTC can reduce the whole blood viscosity( 1s-1,5s-1,50s-1,200s-1)and plasma viscosity, reduce the blood concentration by improving the function of red cell, and it has the effects of anti-coagulation ? anti-thrombosis and anti-platelet aggregation,which may be associated with increasing the content of cAMP, reducing the content of VWF and the platelet membrane glycoprotein CD41/CD61 and 62 P activity. |
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