Studies On The Pharmacodynamics Of SANCHUANGUIDI Granules

Objective: To observe the protective and antioxidative effect of Sanchuan Guidi granules on ischemic myocardium;to explore the mechanism of Sanchua nGuidi granules on hypoxia/reoxygenation(H/R)injury in human umbilical vein endothelial cell line(ECV304)and its effect on the expression of endothelial cells;to investigate the effect of Sanchuan Guidi granule on thehemorheology a nd coagulation function in rats with acute blood stasis;to study the effect of SanchuanGuidi granules on inflammatory response associated cytokines in ather osclerotic rat;and to investigate the effect of SanchuanGuidi granules on the r elease of VW factor and prostacyclin from vascular wall of perfusedumbilical vein.Methods: Wistar rats were divided into 4 groups with 10 in each.Corres ponding dose of SanchuanGuidi granules or normal saline was administered to the rats in each group by gavage and the animals were induced by isoprotere nol(ISO)to establish the myocardial ischemia model 7 days later.24 hours af ter the final administration of ISO,serum contents of SOD,MDA,LDH,and E T as well as myocardial Na+-K+ ATPase,Ca2+-Mg2+ATPase,andtotal ATPase acti vities were measured;cultured ECV304 cells were divided into 4 group sand t reated accordingly.Contents of superoxide dismutase(SOD),glutathione(GSH),and malondialdehyde(MDA)and the expression of nuclear transcription factor–κB(NF-κB)and intercellular adhesion molecule-1(ICAM-1)in each group we re detected;acute blood stasis rat model was induced by cold water bath and subcutaneous injection of ISO and the changes of whole blood viscosity(WB V),plasma viscosity(PV),erythrocyte sedimentation rate(ESR),and hematocri t(HCT)were observed;prothrombin time(PT),activated partial thromboplastin time(APTT),and plasma fibrinogen(FIB)content were determined toinvestig ate the effect Sanchuan Guidi granule on blood coagulation function in blood st asis rats;8-week old male Wistar rats were randomly divided into 5 groups,wh ich are the normal group,the model group,the low-dose Sanchuan Guidi granu le group,the high-dose SanchuanGuidigroup and the atorvastatin group.Levels of inflammatory factors such as tumor necrosis factorα(TNF-α),interleukin1α(I L-1α),and interleukin 6(IL-6)were detected by enzyme linked immunosorbent assay(ELISA);a fresh umbilical cord,a constant flow pump,and a test tube filled with perfusate were assembled to construct a perfusion device,throughwhich perfusatecould be recycled.Perfusates either contained Sanchuan Guidi g ranule or not were used.Supernatant was collected and frozen,and the content s of factor VW and 6-Keto-PGF1αwere determined within 2 weeks.Results: Compared with the model group,serum SOD content in the rats of the Sanchuan Guidi granule groups was significantly increased(p<0.05),ser um MDA,LDH,and ET contents were significantly decreased(p<0.05 or p<0.01),and the activities of Na+-K+ ATPase,Ca2+-Mg2+ATPase,and total ATPase w ere significantly increasedin myocardial tissue(p<0.05 or p<0.01);oxygen free r adical scavenging capacity of the model group,high-and low-dose SanchuanG uidi granule groups was significantly reduced and the expression of NF-κB an d ICAM-1 was outstanding higher than the normal group,among which the ox ygen free radical scavenging capacity of the high-and low-dose SanchuanGuidi granule groups was significantly higher than the model group and the express ion of NF-κB and ICAM-1 was significantly lower than the model group in a dose-dependent manner.Compared with the normal group,model group the he morheology indexes increased significantly,PT and APTT were significantly sh ortened,and the FIB levels increased significantly.Compared with the model g roup,the hemorheology indexes were significantly reduced,the PT and APTT were significantly prolonged,and the content of FIB was significantly decrease d by the administration of the drug in the thigh-and low-dose SanchuanGuidi granule groups;model group myocardial TNF-α,IL-1α,and IL-6 leves were s ignificantly increased,compared with the normal group(p<0.05)difference was statistically significant;the concentration of myocardial TNF-αof the two Sanc huanGuidi granule groups and the atorvastatin group was significantly decrease d(p<0.05);the concentration of myocardial IL-1α in these three groups was redu ced(p>0.05);and the concentration of myocardial IL-6 in these three groups were reduced(p<0.05).After 30 minutes of the perfusion via umbilical vein,VW factor and 6-Keto-PGF1αwere detectable in the perfusate and thrombin o bviously stimulated the release of VW and prostacyclin,of which the release o f VW was inhibited by the application of Sanchuan Guidi granule and that of prostacyclin was not affected.Conclusions: Sanchuan Guidi granules has a protective effect in myocardial ischemia,and the mechanism may involve its antioxidant and the changes of LDH,ET,and ATPase;SanchuanGuidi granule effectively improves the oxygen free radical scavenging capacity of the cells,decreases the activity of NF-κBand ICAM-1,and reduces the H/R injury;Sanchuan Guidi granule significantly improve shemorheology and abnormal coagulation functionin blood stasis rats,providing a basis for the understanding of the scientific connotation of Sanch uan Guidi granule;Sanchuan Guidi granule can reduce the exudation of TNF-αand IL-6,inhibit atherosclerosis inflammation;and Sanchuan Guidigranule-co ntaining perfusatere markably inhibits the release of VW factor,indicating the effects of promoting blood circulation and antithrombosis of Sanchuan Guidi gr anule.