长春西汀对脑缺血再灌注损伤的保护作用及机制

发布时间：2018-06-18 11:51

本文选题：脑缺血再灌注 + 长春西汀　；参考：《吉林大学》2017年硕士论文

【摘要】：目的:探讨长春西汀对脑缺血再灌注损伤的保护作用及机制方法:选用鼠龄3~4个月、体重为270±30 g的健康雄性Wistar大鼠,制作大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)模型,采用缺血2h再灌注12h的时间点。第一部分实验分为5组:Sham组(假手术组),IR组(模型组),Vinp组(长春西汀组)设置不同干预剂量亚组,即Vinp5组、Vinp10组、Vinp20组。采用Longa五分制评分法,对各组动物进行神经功能学评分;TTC染色计算脑梗死面积百分比。第二部分实验分为3组:Sham组、IR组、Vinp10组。采用Western blot检测AQP-4、MMP-9蛋白的表达,分析长春西汀对血脑屏障通透性的影响;检测SOD活性、MDA含量,评价长春西汀抗自由基损伤的作用;采用Western blot检测IL-1β、IL-10、TNF-α蛋白的表达,探讨长春西汀的抗炎作用;采用Western blot检测Bcl-2、Bax及Caspase-3蛋白表达,探讨长春西汀抗细胞凋亡的机制;采用Western blot检测Cx43及p-Cx43蛋白表达水平,明确长春西汀是否通过调控缝隙连接发挥神经保护作用。结果:(1)大鼠脑缺血再灌注后出现明显的神经功能缺损症状,多表现为向右侧倾倒、转圈,或右侧上肢不能完全伸展;Vinp10组、Vinp20组与IR组相比神经功能学评分均明显改善(P0.05),而Vinp10与Vinp20组间无明显差异(P0.05)。(2)与IR组相比,Vinp5组大鼠脑梗死面积未见明显缩小(P0.05),Vinp10及Vinp20组脑梗死面积可见明显减少(P0.01);Vinp20组与Vinp10组相比无明显差异(P0.05)。因此,后续试验中选用10mg/Kg为Vinp给药浓度。(3)脑缺血再灌注后大鼠皮层梗死区AQP-4与MMP-9蛋白表达明显增高(P0.01);Vinp10组与IR组相比可显著降低上述蛋白表达量(P0.01)。(4)脑缺血再灌注后SOD活性明显降低、MDA含量显著升高(P0.01);Vinp10组与IR组相比可显著提升SOD活性、降低MDA含量(P0.01)。(5)脑缺血再灌注后IL-1β、TNF-α蛋白表达明显升高(P0.01)、IL-10蛋白表达未见明显变化(P0.05);Vinp10组与IR组相比明显降低IL-1β、TNF-α蛋白表达并升高IL-10蛋白表达(P0.01)。(6)脑缺血再灌注后Bax及Caspase-3蛋白表达明显升高(P0.01),而Bcl-2蛋白表达、Bcl-2/Bax比值则明显下降(P0.01);与IR组相比,Vinp10可明显降低Bax及Caspase-3蛋白表达(P0.01),并显著升高Bcl-2蛋白的表达及Bcl-2/Bax比值(P0.01)。(7)脑缺血再灌注后Cx43及p-Cx43蛋白表达明显减低(P0.01);与IR组相比,Vinp10组可上调上述蛋白表达量(P0.05)。结论:长春西汀可以显著改善脑缺血再灌注损伤大鼠神经功能缺损、减少脑梗死面积。这种保护作用是通过减轻血脑屏障通透性、清除自由基、抗炎、抗凋亡等机制实现的,而长春西汀对Cx43的调控可能是上述保护机制的基础。
[Abstract]:Objective: to investigate the protective effect and mechanism of vinpocetine on cerebral ischemia-reperfusion injury. Methods: the middle cerebral artery occlusion (MCAO) model was established in healthy male Wistar rats aged 34 ~ 4 months and weighing 270 卤30 g. The time points of ischemia 2 h and reperfusion 12 h were used. The first part of the experiment was divided into 5 groups: group V: Sham (sham-operated group) and group IR (model group): Vinp10 group (Vinp10 group) with different intervention dose subgroup (Vincetine group), or group Vinp10 group (group Vinp20). The percentage of cerebral infarct area was calculated by neurologic score and TTC staining in each group by Longa five-point scoring method. The second part of the experiment was divided into three groups: 1% Sham group, IR group and Vinp 10 group. The effect of vinpocetine on the permeability of blood-brain barrier was analyzed by Western blot, and the effect of vinpocetine on free radical injury was evaluated by detecting the activity of SOD and MDA, and the expression of IL-1 尾 -IL-10 TNF- 伪 protein was detected by Western blot. To investigate the anti-inflammatory effect of vinpocetine, to detect the expression of Bcl-2P Bax and Caspase-3 protein by Western blot, to explore the mechanism of vinpocetine anti-apoptosis, and to detect the expression levels of Cx43 and p-Cx43 by Western blot. It is clear whether vinpocetine plays a neuroprotective role by regulating gap junctions. Results (1) after cerebral ischemia-reperfusion, the rats showed obvious neurological impairment symptoms, most of which showed that they were tilted to the right and turned in circles. Or the right upper limb could not be completely stretched. The neurological functional score of Vinp20 group was significantly improved than that of IR group, but there was no significant difference between Vinp10 and Vinp20 group (P0.05. 2) compared with IR group, there was no significant reduction of cerebral infarction area in Vinp10 group and Vinp20 group (P0.05 Vinp10 group and Vinp20 group). There was no significant difference in infarct size between Vinp20 group and Vinp10 group. therefore In the follow-up test, 10 mg / kg was chosen as Vinp administration concentration. 3) the expression of AQP-4 and MMP-9 in cerebral cortex infarction area was significantly increased after cerebral ischemia and reperfusion. Compared with IR group, the expression of AQP-4 and MMP-9 in Vinp10 group was significantly lower than that in IR group. Compared with IR group, the activity of SOD was significantly increased in the group of P0.01C Vinp10, the content of MDA was significantly decreased, and the content of MDA was significantly increased. Decrease of MDA content and expression of IL-1 尾 -TNF- 伪 after cerebral ischemia-reperfusion; increased expression of IL-1 尾 -TNF- 伪 after cerebral ischemia-reperfusion. No significant change in the expression of IL-10 protein in P0.05TNF- 伪 and increased expression of IL-10 protein after cerebral ischemia-reperfusion compared with IR group) Bax and Caspase-3 proteins in Vinp10 group after cerebral ischemia-reperfusion were significantly lower than those in IR group (P 0.01. 0. 6) the expression of IL 1 尾 TNF- 伪 protein and the expression of IL 10 protein increased after cerebral ischemia reperfusion. Compared with IR group, Vinp10 significantly decreased the expression of Bax and Caspase-3 protein, and increased the expression of Bcl-2 protein and Bcl-2 / Bcl-2 / Bax ratio (P0.01. 7) after cerebral ischemia-reperfusion (Cx43 and p-Cx43 protein surface) after cerebral ischemia-reperfusion, Vinp10 significantly decreased the expression of Bax and Caspase-3 protein, and increased the expression of Bcl-2 protein and Bcl-2 / Bax ratio (P0.01. 7) after cerebral ischemia-reperfusion, Vinp10 significantly decreased the expression of Bax and Caspase-3 protein and increased the expression of Bcl-2 protein and Bcl-2 / Bax ratio (P0.01. 7). Compared with IR group, Vinp10 group could upregulate the expression of the above protein. Conclusion: vinpocetine can significantly improve the neurological deficit and reduce the area of cerebral infarction in rats with cerebral ischemia reperfusion injury. This protective effect is achieved by reducing blood-brain barrier permeability, scavenging free radicals, anti-inflammatory, anti-apoptosis and other mechanisms. The regulation of vinpocetine on Cx43 may be the basis of the above protective mechanism.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.3

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