Protective Effects And Mechanism Of HSYA And Boswellic Acid On Cerebral Ischemia Reperfusion And Blood Stasis Syndrome

Background and objectiveAccording to the culmination of surgery(Wai Ke Da Cheng),Carthamus tinctorius L and Boswellia serrate(B.serrata)are described as a combination,famous as Huoxue Dingtong Decoction in clinical practice,which appears to be effective in the treatment of coronary heart disease and angina pectoris with symptom of heart blood stasis.Carthamus tinctorius L has the function of activating blood circulation to dissipate blood stasis,and Boswellia serrate has the function of promoting qi,restoring menstrual flow,detumescence and detoxification.Preliminary research shows that KBA can improve cerebral ischemia reperfusion,and beta-boswellic acid can improve rat blood stasis.Hydroxy safflower yellow A(HSYA)also has the effects of promoting blood circulation to remove blood stasis and improving cerebral ischemia reperfusion injury,however,few studies have been done on the combination use of HSYA and KBA or HSYA and ?-BA.Therefore,this paper topic is prepared from two aspects to research the synergistic effects of safflower and boswellic acid.The first is to study the mechanism of HSYA combined with KBA to protect the nerve injury induced by cerebral ischemia reperfusion,the second aspect is to study the mechanism of HSYA combined with ?-BA improving vascular endothelial injury induced by blood stasis syndrome.Part 1 HSYA and KBA improve the neurological damage induced bycerebral ischemia reperfusion Purpose1.To investigate whether HSYA and KBA can improve nerve injury induced by cerebral ischemia reperfusion and the synergistic mechanism.2.To determine synergistic effects of HSYA and KBA on the oxidative stress of PC12 cells induced by oxygen glucose deprivation in vitro.MethodsThe focal cerebral ischemia-reperfusion model was induced by the middle cerebral artery occlusion(MCAO).The changes of cerebral infarction area were observed after the injection of HSYA and KBA.In vitro,the damage model of PC12 cells induced by oxygen glucose deprivation(OGD)was used to detect the changes of malondialdehyde(MDA)and superoxide dismutase(SOD)in antioxidant system.Results1.HSYA(100 mg/kg),KBA(25 mg/kg)and HSYA(50 mg/kg)+KBA(12.5 mg/kg)can improve cerebral infarct size and neurological score in rats with cerebral ischemia reperfusion.In addition,the combination group of HSYA(50 mg/kg)+KBA(12.5 mg/kg)is more effective(P<0.05).2.MTT results showed that HSYA(10 ?M),KBA(50 ?M)and HSYA(5?M)+KBA(25?M)have anti apoptosis effect of PC12 induced by OGD,HSYA(5 ?M)and KBA(25 ?M)had significant anti apoptotic(P<0.05).ConclusionIt was confirmed that the combination of HSYA and KBA had synergistic effect on improving the nerve injury induced by cerebral ischemia reperfusion injury,also HSYA and KBA anti apoptotic and oxidative of PC12 cells induced by OGD stress synergistically.Part 2 HSYA and ?-BA improve vascular injury induced by blood stasis syndrome Purpose1.To study whether the combination of HSYA and ?-boswellic acid(?-BA)can improve the abnormal hemorheology of rats with blood stasis syndrome and protection of vascular endothelial function,and figure out if HSYA and ?-boswellic are synergistic?2.To investigate the protective effects of HSYA and ?-BA on the injury of HUVEC cells induced by OGD and the influence on NO formation,to study whether the protective effect is synergistic.MethodsIn vivo studies,the rat of blood stasis syndrome was used.After the injection of HSYA and ?-BA,the whole blood viscosity and coagulation function,the concentration of ET-1 and NO in plasma was detected to observe the effect on hemorheology of rats and the changes of vascular endothelial function.The pathological changes were observed by carotid artery.HUVEC cells induced by OGD were used in vitro study,after giving HSYA and ?-BA,the effect on cell activity and NO function was detected.Result1.Compared with the Control group,the whole blood viscosity of the model group increased significantly,activated partial thromboplastin time(APTT),thrombin time(TT)and prothrombin time(PT)was significantly shortened,fibrinogen(FIB)content and plasma levels of ET-1 was increased,the level of NO was decreased.Compared with the model group,the whole blood viscosity of HSYA(100 mg/kg),?-BA(100 mg/kg)and HSYA(50 mg/kg)+ ?-BA(50 mg/kg)significantly decreased,TT,APTT and PT were significantly prolonged(P < 0.05),the content of fibrinogen reduced(P<0.05),the plasma ET-1 level decreased,plasma levels of NO increased(P<0.05).In addition,the combination group of HSYA(50 mg/kg)+ ?-BA(50 mg/kg)was more significant(P<0.05).2.The results of MTT showed that HSYA and ?-BA has anti-apoptotic effect on HUVEC induced by OGD.The anti-apoptotic effect of HSYA(10 ?M),?-BA(50 ?M)and HSYA(5 ?M)+?-BA(25 ?M)on HUVEC cells was significant(P<0.05),and the effect of combination group of HSYA(5 ?M)and ?-BA(25 ?M)was stronger(P<0.05).3.HSYA(10 ?M),?-BA(50 ?M)and HSYA(5 mol/L)and ?-BA(25 mol/L)can increase HUVEC cells to generate NO(P<0.05),and the effect of combination group of HSYA(5?M)and ?-BA(25?M)was stronger(P<0.05).ConclusionIt was confirmed that there is a synergistic effect of HSYA and ?-BA on improving hemorheology induced by blood stasis syndrome,there is also a synergistic effect of protecting vascular endothelial cells.The synergistic effects on protecting the HUVEC cell induced by OGD and enhancing the NO generation was also confirmed.