ESR Study And Antioxidation By Water Extract Of Carthamus Tinctorius L.

Many researches showed that oxygen free radicals in body are directly related to more than 100 kinds of human diseases.Normally the oxidation and anti-oxidation are in balance. When the body suffers the attack of physical or chemical factors,or other certain factors,it produces a lot of oxygen free radicals,leading to the imbalance and oxidation stress,and raises a number of diseases.Therefore,prevention of oxygen free radicals from damage of body,and reducing the incidence of the disease are a key problem which people have been looking forward to solving.Carthamus tinctorius L.as a Chinese medicine was commonly used for promoting blood circulation and modern pharmacological researches was found that a lipid-lowering and anti-oxidation role in the prevention and treatment of cardiovascular diseases.Therefore,this study was to investigate the effect of water extract of Carthamus tinctorius L.on the oxidative injury in order to provide the scientific evidence for wide clinical application.In this study the model of hyperlipidemia rats,oxidized low dentisy lipoprotein(ox-LDL) induced injury in rat cardiac microvascular endothelial cell,hypoxia reoxygenation induced injury in cardiac microvascular endothelial cells and cardiomyocytes,hydrogen peroxide (H₂O₂) induced injury in cardiomyocytes were used to observe the antioxidant effects of water extract of Carthamus tinctorius L.and the the mechanism.The results were as follows:1.Antioxidant effects of water extract of Carthamus tinctorius L.in hyperlipidemia rats Antioxidant effects of Water extract of Carthamus tinctorius L.were observed by feeding the high-lip food for establishing hyperlipidemia rats model.The rats were divided into five groups:control group,hyperlipidemia model group,Water extract of Carthamus tinctorius L. (4.05,2.03,0.81 g crude drug·kg^-1) dose group.The rats were fed with hypercholesterol diet except control group.After four weeks,blood was collected to measure the level of blood lipid,Malondialdehyde(MDA),superoxide dismutase(SOD) and nitric oxide(NO).The results demonstrated that the water extract of Carthamus tinctorius L.decreased the levels of blood lipid.The rise of MDA was attenuated and the activity of SOD significantly increased, which comparing with the model group.2.Antioxidant effects of water extract of Carthamus tinctorius L.on ox-LDL induced injury in cardiac microvascular endothelial cellsBy using the third generation of rat cardiac microvascular endothelial cells(rCMEC),the protection of water extract of Carthamus tinctorius L.were investigated after ox-LDL (100μg·mL^-1) induced damage.The supernatant was collected for detecting lactate dehydrogenase(LDH),MDA,SOD,xanthine oxidase(XOD),glutathione peroxidase (GSH-Px),nitric oxide(NO),nitric oxide synthase(NOS) activity,and cell suspension was collected for detecting oxygen derived free radicals(OFR) by electron spin resonance(ESR). The results demonstrated that water extract of Carthamus tinctorius L.increased the rCMEC survival rate,reduced LDH,MDA and XOD levels,and improved SOD,GSH-Px and NOS activity,while in the cell suspension OFR signal decreased significantly.3.Antioxidant effects of water extract of Carthamus tinctorius L.on hypoxia and reoxygenation injury in rat cardiac microvascular endothelial cells and cultured neonatal rat cardiomyocytesAntioxidant effects of water extract of Carthamus tinctorius L.were studied in the third generation of rCMEC and the cultured neonatal rat cardiomyocytes which induced by hypoxia and reoxygenation injury.The supernatant was collected for detecting LDH,MDA,SOD, XOD,GSH-Px,NO,NOS activity and cell suspension was collected for detecting OFR by ESR.The results demonstrated that water extract of Carthamus tinctorius L.increased the rCMEC and cardiomyocytes survival rate,reduced LDH,MDA and XOD levels,and improved SOD,GSH-Px and NOS activity,while in the cell suspension OFR signal decreased significantly.4.Antioxidant effects of water extract of Carthamus tinctorius L.in cultured neonatal rat cardiomyocytes induced by H₂O₂In cultured neonatal rat cardiomyocytes,the antioxidant effect of water extract of Carthamus tinctorius L.was investigated after H₂O₂(1mmol·L^-1) induced damage.The supernatant was collected for detecting LDH,MDA,SOD,XOD,GSH-Px,NO,NOS activity and cell suspension was collected for detecting OFR by ESR.The results demonstrated that water extract of Carthamus tinctorius L.increased the spontaneous beating and survival rate following H₂O₂ injury.The rise of LDH,MDA and XOD was attenuated,and the activity of SOD,GSH-Px and NOS activity significantly increased,while in the cell suspension OFR signal decreased significantly.In conclusion,antioxidant effects of water extract of Carthamus tinctorius L.were demostrated in the intact animal models,and the same protection was found in ox-LDL induced injury,hypoxia/reoxygenation and H₂O₂ induced injury in vitro model.Therefore, the results were suggested that water extract of Carthamus tinctorius L.had anti-oxidation. The mechanism may be related with scavenging of free radicals,enhancing its clearance, enhancing endogenous antioxidant activity.