Effect Of Compatibility Of Four Kinds Active Ingredients Of Astragalus Membranaceus And Panax Notoginseng On Oxidative Stress And Signal Transduction After Cerebral Ischemia Reperfusion In Mice

Objective:To investigate the influence of astragaloside Ⅳ (the effective component of Astragalus) combined with ginsenoside Rg1, ginsenoside Rb1, notoginsenoside R1(the effective components of Panax notoginseng) on oxidative stress after cerebral ischemic-referfusion in mice and to study its mechanism from Nrf2/HO-1.Methods:By the previous work to get astragaloside Ⅳ(40mg/kg) combined with ginsenoside Rgl(50mg/kg), ginsenoside Rb1(40mg/kg), notoginsenoside R1(10mg/kg), then, models of cerebral ischemia-reperfusion injury were prepared by bilateral common carotid artery ligation for20min followed by1h and24h reperfusion. Brain tissue was taken to yield a10%homogenate, to detect the contents of Malonaldehyde (MDA), Nitric Oxide(NO), Glutathione (GSH), the activity of Superoxide Dismutase (SOD) and T-AOC in brain tissue after reperfusion. The total motor scores (TMS) observed after24h reperfusion brain tissue, number of the survival rate of nerve cells in the CA1region of the hippocampus were assessed by HE staining. The protein content of HO-1and Nrf2(cytoplasm and nucleolus) were determined in brain tissue.Result:Combination between Astragaloside Ⅳ, ginsenoside Rgl, ginsenoside Rbl and notoginsenoside R1has a protective effect on cerebral ischemia-reperfusion model.Significantly the early oxidative stress injury after cerebral ischemia-reperfusion in mice, Compared with the sham group, the ability of AOC, SOD activity and GSH content was decreased in the model group (P<0.01). Compared with the model group, Astragaloside IV. Rg1、Four active components combination. Astragaloside Ⅳ+Rg1、Astragaloside Ⅳ+Rbl were significantly increased GSH contents (P<0.05or P<0.01); Rg1. R1. Four active components combination. Astragaloside Ⅳ+Rgl. Astragaloside Ⅳ+Rb1、 Astragaloside IV+R1were significantly increased T-AOC(P<0.05or P<0.01); Compared with the four active components combination, Rg1、 Rbl and R1can notably increase SOD activity (P<0.05or P<0.01); Compared with the four active components combination, Astragaloside IV. Rbl were significantly increased T-AOC activity(P<0.05or P<0.01); Compared with the four active components combination, Astragaloside Ⅳ、Rg1、Rb1、R1and Astragaloside Ⅳ+R1were significantly increased GSH contents (P<0.01)。 by24h reperfusion after,Compared with the sham group, the contents of MDA. NO significantly were increased (all P<0.01), SOD activity and GSH level were decreased in the model group (P<0.01or P<0.05). Compared with the model group,astragaloside R’ significantly inhibited the increase in MDA, NO (all P<0.01), was less influenced on SOD, GSH (P>0.05); ginsenoside Rgl reduced obviously NO content, had no marked effects on MDA, SOD, GSH (P>0.05); No significant effects (P>0.05) were all found on MDA, NO, SOD, GSH in ginsenoside Rb1and notoginsenoside Rl.Astragaloside Ⅳ combined respectively with the active components of Panax notoginseng (two active components combination) all reduced obviously the contents of MDA and NO (P<0.01or P<0.05), and, the effects in astragaloside Ⅳ+ginsenoside Rbl and astragaloside Ⅳ+notoginsenoside R1were better than those in ginsenoside Rbl, notoginsenoside Rl alone; however, two active components combination had all no effects on SOD activity and GSH level (P>0.05).Four active components combination decreased significantly the contents of MDA and NO, increased significantly SOD activity and GSH level (all P<0.01), furthermore,the effects were more obvious than those in the single-drug and two active components combination (P<0.01or P<0.05). Astragaloside Ⅳ, two or four active components combination decreased MDA, which did not been find in the active components of Panax notoginseng alone.Astragaloside Ⅳ, ginsenoside Rgl, two or four active components combination could inhibit the produce of NO. Four active component alone, two active components combination had no marked effects on SOD and GSH, but which were increased obviously in four active components combination. The influence of astragaloside Ⅳ (the effective component of Astragalus) combined with ginsenoside Rgl, ginsenoside Rbl, notoginsenoside R1(the effective components of Panax notoginseng) were restrain the number of the survival rate of nerve cells in the CA1, then Astragaloside IV combined respectively with the active components of Panax notoginseng (two active components combination) all restrain reduced obviously the number of the survival rate of nerve cells, and, the effects in astragaloside Ⅳ+ginsenoside Rbl and astragaloside IV+notoginsenoside R1were better than those in ginsenoside Rbl, notoginsenoside R1alone. The results of Western-bloting:by24h reperfusion after, therefore its nuclear translocation and the contents of HO-1increases. Four active components combination astragaloside Ⅳ+Rg1、astragaloside Ⅳ+Rb1、astragaloside Ⅳ+R1can accelerate nuclear translocation and the contents of HO-1, and four active components combination、astragaloside Ⅳ+Rg1、astragaloside Ⅳ+Rb1、astragaloside Ⅳ+R1of nuclear translocation and the contents of HO-1were better than those in Four active components combination alone. Experiment results show by24h reperfusion after, in order to adapt stress-stimuli such as ischemia and hypoxia, the brain tissue Nrf2/HO-1pathway to activate the Nrf2nuclear translocation and HO-1expression is increased, thus combating the brain tissue damage caused by ischemia and hypoxia, which is the protection of the body to producereaction. Four active components combination activated Nrf2/HO-1pathways, thus to combating cerebral ischemia and reperfusion brain damage.Conclusion:Combination between Astragaloside Ⅳ, ginsenoside Rgl, ginsenoside Rbl and notoginsenoside R1has a protective effect on cerebral ischemia-reperfusion model, and four active components combination can enhance the protective effect on brain, These active ingredients and their combination restrained the formation of oxygen-derived free radicals after cerebral ischemia reperfusion, and inhibit lipid peroxidation, protection of the activity of antioxidant substance, thus oxidative stress injury against cerebral ischemia. The mechanism maybe that to activate Keapl/Nrf2/HO-1signal pathway, to promoted formation of Nrf2and nuclear translocation, So as to promote the expression of downstream antioxidantgenes such as HO-1the role of oxidative damage.