The Treatment And Mechanism Of Astragalus Membranaceus To CVS As SAH In Rabbit Mode

Part oneCerebral Vasospasm (Cerebral Vasospasm, CVS) is one of the most common and most dangerous complications after spontaneous subarachnoid hemorrhage (SAH),CVS as SAH’s continuous progress will eventually lead to Cerebral ischemia, Cerebral infarction, even so, the CVS as SAH becomes a main cause of death or disability. Related animal experiments and clinical research on CVS as SAH pathogenesis has become a hot area to constantly breakthroughs in the world’s neurosurgical field.Study shows,delayed CVS as SAH is a process involving a multi-factor, and the participation of many links, which including vascular endothelial cell injury, hemoglobin metabolites caused by vascular inflammation, apoptosis, oxidative stress, immune response and gene, racial differences, and many other factors, and among them, vascular inflammation and associated inflammation factors is so closely related the mechanism and plays a fundamental role. On the process of the occurrence and development of CVS after SAH, inflammation factors such as Interleukin lbeta (IL-lbeta)/Endothelin1(ET-1) was found and established as a closely related inflammatory factors to CVS as SAH.Objective1. Evaluating of Animals’ Neurobehavior on CVS as SAH Rabbit Model by Animal Neurobehavioral Score (ANS), and combined with the monitoring results on animals’ cerebral basilar artery (CBA) blood flow velocity (BFV) by Transcranial Doppler ultrasound (TCD), determine whether the CVS as SAH Rabbit Mode is successfully established or not.2.Determinating of contents variation of IL-lbeta/ET-1in rabbits’ cerebrospinal fluid (CSF) on CVS as SAH Rabbit Mode.3.Rreasoning the correlation between inflammation factors IL-lbeta/ET-1and incidence of CVS as SAH.Method1.40NewZealand Rabbits (Male&Female unlimited) Selected, were randomly divided into:SAH Group (n=30), CVS as SAH Rabbit Mode was established by "Blood Secondary Injection of Cisterna Magna", and intrathecal cannuling;Shame Operation Group (n=10), applying the same operation method, Sterile Saline Solution was injected secondary into Cisterna Magna on the same dose.2.Recording ANS Scores, TCD Results on BFV of CBA of all the experimental animals on DO,D1,D4,D7and D9.3.Measuring the contents of IL-lbeta/ET-1in CSF of all the experimental animals on DO,D1,D4,D7and D9by Double Antibody Sandwich Method (ELISA), Comparison and analysis results determined by two groups animals.Results1.None of animals of Shame Operation Group was dead during DO to D9, eating and physical activity was good,while most animals of SAH Group appeard somehow nerve function obstacle, such as paralysis, head to the side dump, even no spontaneous movement and so on after CVS as SAH Rabbit Mode successfully established,14were dead during DO to D9and the Mortality is46.67%. Comparing the animals ANS between SAH Group and Shame Operation Group at same time, and making within-group comparisons on the SAH Group at different time, all datas was statistical significance (P<0.01),except for the datas on D7and D9.2. TCD test Results on BFV of rabbits’ CBA of Shame Operation Group during DO to D9did not show that BFV did not appear obviously changes,while the animals of SAH Group appeared rising trend day by day during DO to D9, of which the most is on D7, and after that, a declining trend revealed. Comparing the TCD Results between SAH Group and Shame Operation Group at same time,the datas owns statistical significance (P<0.05) on D1/D7,and the datas was obviously statistical significance (P<0.01); Making within-group comparisons on the SAH Group at different time, compared with DO (P<0.01), compared with D1(P<0.01), compared with D4(P<0.01).3.The contents of IL-lbeta/ET-1in CSF of experimental animals of Shame Operation Group during DO to D9did not show apparently rise, While the animals of SAH Group appeared rising trend day by day during DO to D9, of which the most is on D7,and after that, a declining trend revealed. Comparing the contents of IL-lbeta/ET-1in CSF of experimental animals between SAH Group and Shame Operation Group at same time,the datas owns statistical significance (P<0.01) on D4/D7/D9of IL-lbeta,on D4/D7of ET-1; Making within-group comparisons on the SAH Group at different time, compared with DO (P<0.01), compared with D1(P<0.01), compared with D4(P<0.01).4.Conbined with the TCD Results and the changes of content of IL-lbeta/ET-1in CSF of experimental animals of SAH Group, curve is drawn in "TIME" as the axis, the BFV of rabbits’CBA and the changes of content of IL-lbeta/ET-1in CSF share similar wave,and both reach to the peak on D7. Statistical correlation analysis was carried out on the datas, the result shows that the changes of content of IL-lbeta/ET-1in CSF of experimental animals and the TCD Results BFV of rabbits’ CBA, respectively, has obvious positive correlation (r=0.931, P<0.01and r=0.951, P<0.01), while the Shame Operation Group animals failed to such a correlation.Conclusion1.CVS as SAH Rabbits Mode established by "Blood Secondary Injection of Cisterna Magna" could successfully simulate the process of CVS as SAH similarly as human being, and dynamic detection the FBV of CBA of rabbits’ by TCD could obtain more accurate evidence of CVS as SAH.Apparent animal neurobehavioral function defect was easy to observed and recorded by animal mode established successfully, therefore, CVS as SAH Rabbits Mode was justified as a better optional animal model to study CVS as SAH of human being.2.CSF specimens of the experimental animal mode collected, analysis of CSF IL-lbeta/ET-1in change, considering the change on FBV of CBA when CVS as SAH happened, comes to conclusions that the difference has significant positive correlation. The experimental results was furtherly verified that vascular inflammation and expression of inflammatory cytokines has a clear correlation with the incidence of CVS as SAH. The changes of content of IL-lbeta/ET-1in CSF could be considered as a very important index to diagnosis and outcome evaluation. Part TwoThe patients’ prognosis of CVS as SAH is involved by a common multiple, complicated factors, and the long-term prognosis of CVS as SAH depends on the differences on the treatment measures on patients,and also,the long-term’s outcome of the patients in CVS as SAH is quite changed a lot. In the field of clinical drug treatment of CVS as SAH,drugs mainly includs Calcium Channel Blockers (CCB),such as Nimodipine; Nonspecific Vasodilator, such as Papaverine;Oxygen Free Radical Scavenger, such as Edaravone and so on, besides,nimo has been evaluated as "a gold therapeutic agents in the treatment of CVS "in the industry. In Chinese medicine treatment of CVS as SAH,there was few new drugs.Modern Practical Chemistry and Pharmacology study found that Chinese traditional medicine Astragalus hoangtchy could play a significant pharmacological effects on anti-inflammatory, spasmolysis, hemangiectasis and antiapoptosis, Astragalus membranaceus can also remove oxygen free radicals and reduce lipid peroxidation and inhibit platelet accumulation,in order to effectively improve the microcirculation. This article introduces the domestic pharmaceutical preparations the Astragalus membranaceus injection for the treatment of CVS as SAH rabbit model,and comparing the therapeutic effect itself with Nimo ingredient medicine—Nimotop, try to explore new ideas on clinical drug treatment on CVS as SAH of human being.Objective1.Comparision and analysis the the treatment effects on Astragalus membranaceus injection and Nimotop injection.2.1nvestigating whether Astragalus membranaceus injection works on SAH’s therapeutic effect is related to effect on inhibition of intracranial and perivascular inflammatory cytokines IL-lbeta/ET-1expression after SAH or not.Method1.45NewZealand Rabbits (Male&Female unlimited) Selected, were used to established CVS as SAH Rabbit Mode by "Blood Secondary Injection of Cisterna Magna", and then randomly divided into:Astragalus membranaceus Group (n=15),Astragalus membranaceus injection3ml(mg)/once by Intraperitioneal way,twice a day; Nimotop Group(n=15),Nimotop injection0.5mg(3ml)/once by Intraperitioneal way,twice a day;Control Group(n=15), Sterile Saline Solution injection3ml/once, by Intraperitioneal way,twice a day,and all the treatments last for9days.2. Recording ANS Scores, TCD Results on BFV of CBA of all experimental animals onD0,Dl,D4,D7and D9.3. Measuring the contents of IL-lbeta/ET-1in CSF of all the experimental animals on DO,D1,D4,D7and D9by Double Antibody Sandwich Method (ELISA),Comparison and analysis results determined by three groups animals.Results1. All animals (n=45) in three groups appeard somehow nerve function obstacle, such as paralysis, head to the side dump, even no spontaneous movement and so on after CVS as SAH Rabbit Mode successfully established. In Control Group,9were dead during DO to D9and the Survival is40.00%;6were dead in Astragalus membranaceus Group and the Survival is60.00%;5were dead in Nimotop Group and the Survival is66.67%.Comparing the animals ANS among three groups animals at same time, and making within-group comparisons at different time, all datas was statistical significance (P<0.05),except for the datas on D7and D9of SAH Group.2.TCD test Results on BFV of rabbits’CBA of three groups during DO to D9appeared rising trend day by day during DO to D9and Control Group arised mostly,prompting that medicines could ease the rising of BFV of CBA following SAH. Comparing the TCD Results among three groups animals at same time,the datas owns statistical significance (P<0.01) on D1/D4,Making within-group comparisons on the SAH Group at different time, compared with DO,D1/D7(P<0.01), compared with D1,D7(P<0.01), compared with D4,D7/D9(P<0.01).3.The contents of IL-lbeta/ET-1in CSF of experimental animals of three groups during DO to D9appeared rising trend day by day during DO to D9.Comparing the contents of IL-lbeta/ET-1in CSF of experimental animals between Control Group and Astragalus membranaceus Group at same time,the datas owns statistical significance (P<0.01) on D1of IL-lbeta,on D4/D7of ET-l,and the datas owns statistical significance (P<0.05) on D4of IL-lbeta,D1of ET-1; Comparing the contents of IL-lbeta/ET-1in CSF of experimental animals between Control Group and Nomotop Group at same time, all datas was statistical significance (P<0.01) of ET-1,except for the datas on DO of SAH Group, while IL-lbeta failed to it.Conclusion1.Application Atragalus membranaceus injection treatment of CVS as SAH in rabbit, Can obviously alleviate CVS on cerebral basilar artery, much more than the experimental animals of Control Group,especially in Neurobehavioral Function Defect(NFD) including eating,physical activity and so on.Besides, the Survival significantly increased. The herapeutic effect of Astragalus membrane is similar as Nimotop, and clinical applications may be considered.2. The higher degree of IL-1beta and ET-1in CSF of the experimental animals in Astragalus membranaceus Group is less than Control group and Nimotop Group, the inference, it is closely related to the aspects of the drug pharmacological effects itself,such as anti-inflammatory, spasmolysis&expansion tube, protecting vascular endothelial cells, inhibiting synthesis and releasing vascular endothelin, improving local blood supply.