药物干预对大鼠急性脑缺血再灌注损伤神经元的保护作用

发布时间：2018-05-09 02:19

本文选题：缺血再灌注损伤 + 缺血半暗带　；参考：《广西医科大学》2013年硕士论文

【摘要】：目的：观察大鼠急性脑缺血再灌注损伤后,缺血半暗带脑组织Na+-K+-ATP酶活性的动态变化,以及脑蛋白水解物和银杏叶提取物对该酶活性的影响,探讨脑蛋白水解物及银杏叶提取物联合应用对大鼠缺血半暗带神经元的保护作用及机制。方法：将600只雄性Wistar大鼠通过数字表法随机分为对照组、蛋白组、银杏叶组、联合组,共4组,每组大鼠根据缺血后再灌注时间的不同又分为缺血2h再灌注6h、24h、48h、72h、7d五个亚组,其中每个亚组大鼠为30只。用10%水合氯醛350mg/kg腹腔注射麻醉大鼠,参照Longa等介绍的栓线法制备大鼠缺血2h再灌注模型,采用Zea Longa评分法判定动物模型与否成功。除对照组外,其余三组均于大鼠造模后5h至断头取脑时每日用药物进行干预一次。脑蛋白组:5ml/kg脑蛋白水解物注射液,银杏叶组：100mg/kg银杏叶提取物注射液,联合组：5ml/kg脑蛋白水解物注射液及100mg/kg银杏叶提取物注射液。各亚组大鼠分别于缺血再灌注6h,24h,48h,72h及7d断头取脑,观察缺血半暗带脑组织Na+-K+-ATP酶活性、脑组织水肿程度、脑梗死范围及神经症状评分的变化。结果： 1.6h时对照组大鼠脑缺血半暗带神经元Na+-K+-ATP酶的活性为(4.810±0.426) U/mg,随着缺血再灌注时间的延长,Na+-K+-ATP酶的活性逐渐降低,于48h达最低值(3.537±0.415) U/mg,72h稍有回升(3.671+0.501) U/mg,7d趋于稳定(5.022±0.623) U/mg。值得注意的规律是,随着缺血再灌注时间的推移,脑组织缺血半暗带神经元Na+-K+-ATP酶活性呈下降趋势,与此同时,脑组织含水量及梗死面积却在逐渐增加。 2.6h时,对照组、蛋白组、银杏叶组及联合组四组之间Na+-K+-ATP酶活性差异无统计学意义(P=0.864),随着缺血再灌注时间的延长,对照组Na+-K+-ATP酶的活性逐渐降低,而蛋白组、银杏叶组及联合组Na+-K+-ATP酶活性较对照组明显升高,差异具有统计学意义(24h、48h及72h亚组,P值均为0.000),蛋白组与银杏叶组之间差异无统计学意义(P0.05)。与单一用药组(蛋白组或银杏叶组)比较,联合组能明显提高大鼠缺血半暗带神经元Na+-K+-ATP酶活性(48h、72h亚组,P值均为0.000),与此同时,还能显著降低脑组织含水量(24h、48h、72h亚组,P值均为0.000),减小脑梗死面积(24h、48h、72h、7d亚组,P值均为0.000),减少神经症状评分(48h、72h、7d亚组,P值均为0.000)。结论： 1.急性脑缺血再灌注损伤后缺血半暗带脑组织Na+-K+-ATP酶活性呈动态变化； 2.Na+-K+-ATP酶活性变化与脑组织水肿程度、脑梗死面积大小密切相关,在缺血半暗带脑组织缺血再灌注损伤过程中发挥着至关重要的作用。 3.联合用药具有协同作用,较单一用药能更有效地抑制缺血级联反应,从而最大限度地挽救缺血半暗带神经元的功能。
[Abstract]:Aim: to observe the dynamic changes of Na K ATPase activity in the brain of ischemic penumbra after acute cerebral ischemia-reperfusion injury in rats, and the effects of brain protein hydrolysate and ginkgo biloba extract on the activity of Na K ATPase. To investigate the protective effect and mechanism of brain protein hydrolysate and ginkgo biloba extract on ischemic penumbra neurons in rats. Methods: 600 male Wistar rats were randomly divided into four groups by digital table: control group, protein group, ginkgo biloba leaf group, combined group. According to the time of reperfusion after ischemia, each group was divided into five subgroups: 2 h ischemia, 24 h reperfusion and 48 h reperfusion for 72 h and 7 d. There were 30 rats in each subgroup. 10% chloral hydrate 350mg/kg was injected intraperitoneally into anesthetized rats. The model of ischemia 2 h reperfusion was made by referring to the method of suture introduced by Longa et al. The Zea Longa scoring method was used to determine whether the model was successful or not. In addition to the control group, the other three groups were treated with drugs once a day, 5 hours after the rat model was made and the brain was removed from the head. Brain protein group: 5 ml / kg brain protein hydrolysate injection, ginkgo leaf group 100 mg / kg Ginkgo biloba extract injection, combined group 5 ml / kg brain protein hydrolysate injection and 100mg/kg ginkgo leaf extract injection. The rats in each subgroup were decapitated for 72 h and 7 d respectively at 6 h, 24 h and 48 h after reperfusion. The changes of Na K ATPase activity, brain edema degree, infarct area and neurological symptom score were observed in the ischemic penumbra. Results: At 1.6 h, the activity of Na -K ATPase was 4.810 卤0.426) U / mg in the neurons of cerebral ischemic penumbra in the control group. With the prolongation of ischemia reperfusion time, the activity of Na -K ATPase decreased gradually, and increased slightly at 48 h to the lowest value of 3.537 卤0.415) U / mg ~ (-1) (3.671 ~ 0.501) U / (mg ~ (-1). The activity of Na ~ (-K) -ATPase increased to 5.022 卤0.623) / 渭 mg / g at 48 h. It is worth noting that the activity of Na-K-ATPase in the neurons of the ischemic penumbra decreased with the time of ischemia and reperfusion, but the water content and the infarct area of the brain tissue increased gradually at the same time. At 2.6 h, there was no significant difference in Na K ATPase activity between control group, protein group, ginkgo biloba leaf group and combination group. The Na K ATPase activity of control group decreased gradually with the prolongation of ischemia reperfusion time, while the activity of Na K ATPase decreased in protein group. The activity of Na -K ATPase in the ginkgo leaf group and the combined group was significantly higher than that in the control group, and the difference was statistically significant (P = 0.000) between the 24 h and 72 h subgroups, but there was no significant difference between the protein group and the ginkgo leaf group (P 0.05). Compared with the single drug group (protein group or ginkgo biloba group), the combined group could significantly increase the activity of Na-K-ATPase in ischemic penumbra neurons. It also significantly decreased the water content of brain tissue (P = 0.000), decreased the area of cerebral infarction (P = 0.000), reduced the area of cerebral infarction (24 h / 48 h, 72 h / 7 d), reduced the neurological symptom score (P = 0.000) and reduced the neurological symptom score (P = 0.000) at 48h and 7h / 7d respectively in the subgroup of 24 h, 48 h, 72 h and 72 h, respectively. Conclusion: 1. After acute cerebral ischemia-reperfusion injury, the activity of Na -K ATPase in the brain tissue of ischemic penumbra showed dynamic changes. The change of 2.Na-K-ATPase activity is closely related to the degree of cerebral edema and the size of cerebral infarction. It plays an important role in the process of cerebral ischemia-reperfusion injury in ischemic penumbra. 3. The synergistic effect of combined medication can inhibit ischemic cascade reaction more effectively than that of single medication, thus saving the function of ischemic penumbra neurons to the maximum extent.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R743.3

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