| Objective:1.Spinal Cord Ischemia/Reperfusion Injury Models was established by blocking rabbit lumbar Artery, and analyed vessel distribution of spinal by vesselcast technique and digital subtraction angiography that demonstrated the spinal blood supply.2.Tanshinone-Ⅱ A of phamacokineties was analysed via Liquid Chromatography-Mass Technology.3. To evaluate the protective effect of Tanshinone-ⅡA and discuss the possible mechanism.Method:1. Experimental animals and groups:90new zealand rabbits which is3-month age weighting2.0±0.2kg were randomly divided into three group:12rabbits in establishment and identifi-cation model group;24rabbits in Tanshinone-ⅡA Pharmacokinetic analysis group;54rabbits in mechanism analysis group.2. Established and demonstrated Spinal Cord Ischemia/Reperfusion Injury Models: Firstly,Two rabbits of stablishment and identification model group was used for establishing models and demonstrating the spinal blood supply;the other rabbits were randomly divided into two group:Blocking abdominal aortic group and lumbar arteries, Surgerial separate the ministry of right renal artery without occlusion of the abdominal aorta after the operation is terminated in the Blocking abdominal aortic group, Blocking lumbar arteries group was separated lumbar arteries,Then contrast agents injected in aorta vessel, acquisition image were taken. Next blocking L3-L6lumbar arteries, repeat injecting contrast agents and acquisition image were taken again.Finally removing artery block, repeat injecting contrast agents and acquisition image were taken again in order to verify the spinal cord ischemia-reperfusion model successful made via vesselcast technique and digital subtraction angiography.3. Tanshinone-ⅡA Pharmacokinetic analysis group were randomly divided into four group:0.5hours group;1hours group;4hours group;8hours group,6rabbits in each group. Every rabbits was collected blank cerebrospinal fluid (CSF) specimen, then injected Sulfotanshinone Sodium Injection along the edge of ear vein, According to the above methods collected Medicated CSF specimen, analysis pharmacokinetics of Tanshinone-IIA through Liquid Mass Spectrometry Technology.4. Test group were randomly divided into three group:6rabbits in sham-operated group,24rabbits in ischemia reperfusion group and24rabbits in Tanshinone-IIA group. Surgerial separate the ministry of L3-L6lumbar arteries after the operation is terminated in the sham group. Using the SSCI improved model without drug intervention before or after surgery in the ischemia reperfusion group, Sulfotanshinone Sodium Injection intraperitoneally half an hour before and after the establishing of SSCI model in Tanshinone-IIA injection group. Each group drawn after modeling0.5h,1h,4h and8h, a total of four time points,6rabbits at each time point. The protein levels of nuclear factor-kappaβ, vascular cell adhesion molecule-1and hemorrheologic indexes were detected. Spinal cord tissue was detected by ELISA, and the positive cells of nuclear factor-kappaB, vascular cell adhesion molecule-1spinal cord tissue was detected by immunohistochemical staining.5. All data were analized via spss16.0software and image processing system,we collect, classify and analyze the data and image.Results:1. The rabbit models of spinal cord ischemia reperfusion were successfully made and identified.2.2and7h Medicated CSF specimen which was injected in Sulfotanshinone Sodium Injection was diferent from blank cerebrospinal fluid cerebrospinal fluid,2h samples’peaks at a retention time of8.92minutes and the Molecular Weight was399.2,7h samples’peaks at a retention time of31.29minutes and the Molecular Weight was419.3. The expression of both NF-κβ and VCAM-1were increasing fast in ischemia-reperfusion Group while they were significantly decreased during SSCI in Tanshinone-IIA (p<0.05).There were VCAM-1-positive blood vessels4hrs after reperfusion, and keep increasing till reperfusion8hrs in Tanshinone-IIA group and significantly reduced than sham-operated group at4h,8h after reperfusion (p<0.05). VCAM-1-positive blood vessels of the Tanshinone-IIA group was especially decreased after reperfusion (p<0.01). It indicated Tanshinone-IIA could effectively change all the indexes of hemorrheologic indexes (p<0.01).Conclusions: 1. It is a good model of spinal cord ischemia-reperfusion injury that has good repeatability, complete ischemia, easy to operate, and can be a very good simulation of the human spinal cord due to various causes ischemia-reperfusion injury in the pathophysiology process, Therefore, it is a benifical model to study the spinal cord ischemia-reperfusion injury,worthy of clinical application.2. Tanshinone IIA could enter cerebrospinal fluid via circulatory system, then it metabolized other substances in cerebrospinal fluid, So as to protect the function of the spinal cord tissue.3. Tanshinone IIA could significantly reduced inflammation that is lead to Ischemia/reperfu-sion injury of Spinal cord,it protects Ischemia/reperfusion injury of Spinal cord via to reduce inflammatory factor NF-κβ, VCAM-1and hemorrheologic indexes. |
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