ABT-702在大鼠脑缺血再灌注损伤中的作用

发布时间：2018-05-03 19:44

本文选题：ABT-702 + 缺血再灌注　；参考：《新乡医学院》2017年硕士论文

【摘要】：背景缺血性脑血管病的发病率近年来呈逐年递升的趋势,给人类身体健康和生存质量带来极大的危害,因而对该病不断深入和细致地研究,成为神经学科科学家研究的热门方向,不断寻求新的治疗靶点,尽早挽救缺血半暗带成为目前缺血性脑血管病研究的重点之一。研究发现腺苷对缺血性脑血管病具有保护作用,ABT-702作为新型的非核苷类腺苷激酶抑制剂(Adenosine Kinase Inhibitors AKI),能够提高体内腺苷(Adenosion Ado)水平,在既往的研究中,研究者们证实了ABT-702在许多人类疾病中具有治疗或保护作用,但是ABT-702在缺血性脑血管病中的作用却暂无文献报道。目的探讨在大鼠脑缺血性再灌注损伤模型中,腺苷激酶抑制剂ABT-702应用后,大鼠大脑脑组织中腺苷含量的变化,以及自噬蛋白Beclin1的表达情况,进一步验证ABT-702对大脑具有保护作用,为其在缺血性脑血管病中的研究及应用提供前沿基础。方法雄性SD大鼠84只,随机分为对照组、假手术组、模型组、ABT-702组四个组,每组21只,所有大鼠在再灌注后均进行大鼠神经功能缺失评分,每组选取6只进行TTC染色检测梗死面积,各组再随机选取9只分为3个亚组,在不同时间点通过高效液相色谱法(High Performance Liquid Chromatography HPLC)检测脑组织腺苷含量,最后各组再随机选取6只通过RT-PCR法检测Beclin1的表达。结果1、ABT-702组神经缺失功能评分低于模型组。通过Zea Longa法进行评分后,对照组和假手术组评分均为0分,模型组与ABT-702组均有神经功能缺失症状,但ABT-702神经功能缺失评分低于模型组,差距具有统计学意义(P0.05)。2、ABT-702组的梗死体积明显低于模型组。对照组及假手术组TTC染色后未见梗死区域,模型组及ABT-702组均可见大片梗死区域(TTC染色后为白色)。但ABT-702组梗死体积与模型组相比,梗死体积小于模型组,差距具有统计学意义(P0.05)。3、HPLC法检测大鼠大脑腺苷含量变化。各组在再灌注后,分别通过HPLC法在不同时间点(2h、6h、12h)进行脑组织中腺苷含量检测。结果提示,各组在各时间点均可以检测处腺苷的存在。对照组与假手术组两组对比,在各时间点腺苷含量水平没有差距,模型组与ABT-702组在再灌注2h、6h时腺苷含量均高于对照组与假手术组组。ABT-702组与模型组对比,ABT-702组在再灌注各时间段腺苷含量水平均高于模型组。但是在再灌注12h后时ABT-702组的腺苷含量水平低于本组再灌注后2h及6h时的水平。4、脑组织中Beclin1 mRNA表达。各组在再灌注6h后,Beclin1 mRNA均有表达,但模型组Beclin1蛋白的表达量高于对照组与假手术组。而ABT-702组的Beclin1表达量明显高于其它三组。结论1、ABT-702在大鼠缺血再灌注损伤中具有保护作用,减轻了大鼠神经功能的损伤,减小了大鼠脑组织的梗死体积;2、ABT-702应用后大鼠脑组织内腺苷含量明显增加,并且在再灌注6h时脑组织腺苷含量最高;3、ABT-702在大鼠缺血再灌注模型中,提高了自噬(Autophagy)相关蛋白Beclin1的表达水平,这可能是ABT-702在大鼠缺血再灌注损伤(Cerebral Ischemia Reperfusion Injury,CIRI)中起保护作用的机制之一。
[Abstract]:The incidence of background ischemic cerebrovascular disease has been increasing year by year, which has brought great harm to the health and quality of life of human being. Therefore, it has become a hot direction in the research of neuroscientists, seeking new therapeutic targets and saving the ischemic penumbra as soon as possible. One of the key points in the study of bloody cerebrovascular disease. The study found that adenosine has a protective effect on ischemic cerebrovascular disease. ABT-702, as a new non nucleoside adenosine kinase inhibitor (Adenosine Kinase Inhibitors AKI), can improve the level of adenosine (Adenosion Ado) in the body. In the previous study, researchers confirmed that ABT-702 is in many people. However, the role of ABT-702 in ischemic cerebrovascular disease is not reported. Objective to explore the changes of adenosine content in the brain tissue of rat brain and the expression of autophagin Beclin1 in rat brain tissue after the application of adenosine kinase inhibitor ABT-702 in the rat model of ischemic cerebral reperfusion injury. One step was to verify the protective effect of ABT-702 on the brain and provide the Frontier Foundation for its research and application in ischemic cerebrovascular disease. Methods 84 male SD rats were randomly divided into control group, sham operation group, model group and group ABT-702, with four groups of 21 rats in each group. All rats were scored in each group after reinjection, each group was selected. The infarct area was detected by TTC staining in 6 rats, and 9 groups were randomly selected to be divided into 3 subgroups, and the content of adenosine in brain tissue was detected by High Performance Liquid Chromatography HPLC at different time points. Finally, 6 rats were randomly selected to detect the expression of Beclin1 by RT-PCR method. Results 1, ABT-702 group nerve deletion. The score of the function score was lower than that of the model group. The scores of the control group and the sham operation group were all 0 points after the Zea Longa method. The model group and the ABT-702 group had the symptoms of neural function deletion, but the score of the ABT-702 neural function loss was lower than the model group, the gap was statistically significant (P0.05).2, and the infarct volume in the group ABT-702 was significantly lower than that of the model group. The infarct area was not found in the sham operation group after TTC staining. The infarct area in the model group and the ABT-702 group were all visible (white after TTC). But the infarct volume in the ABT-702 group was less than the model group, and the infarct volume was statistically significant (P0.05).3. The HPLC method was used to detect the changes in the cerebral adenosine content in the rats. Each group was divided after reperfusion. Do not use HPLC method to detect adenosine content in brain tissue at different time points (2h, 6h, 12h). The results suggest that each group can detect the presence of adenosine at all time points. Compared with the two groups of sham operation group, the level of adenosine content is no difference at all time points, and the adenosine content in the model group and the ABT-702 group is higher than that of the 2H and 6h. In group.ABT-702 and sham group, the level of adenosine content in group ABT-702 was higher than that of model group at all time of reperfusion, but the level of adenosine content in group ABT-702 was lower than that of 2H and 6h after reperfusion after reperfusion of 12h, and the expression of Beclin1 mRNA in brain tissue. After reperfusion for 6h, Beclin1 mRNA was all after reperfusion. The expression of Beclin1 protein in the model group was higher than that of the control group and the sham operation group. The expression of Beclin1 in the ABT-702 group was significantly higher than that of the other three groups. Conclusion 1, ABT-702 has protective effect in the rat ischemia reperfusion injury, alleviates the injury of the nerve function and reduces the infarct volume of the rat brain tissue; 2, ABT-702 after application. The content of adenosine in the rat brain was significantly increased and the content of adenosine in the brain was highest at 6h reperfusion. 3, ABT-702 increased the expression of autophagy (Autophagy) related protein Beclin1 in the rat model of ischemia-reperfusion, which may be the protection of ABT-702 in the ischemia-reperfusion injury (Cerebral Ischemia Reperfusion Injury, CIRI) in rats. One of the mechanisms of protection.

【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.3

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