钠钙交换体1、3参与远隔创伤后处理的脑保护作用
发布时间：2018-12-11 15:34
【摘要】：目的：应用大鼠局灶性脑缺血再灌注模型研究远隔创伤后处理(remote postconditioning of trauma, Tra-Post)对大鼠脑缺血/再灌注损伤的影响,研究钠钙交换体(sodium-calcium exchangers, NCX)在这种后处理方式中的作用。 方法：45只雄性SD大鼠随机分成3组(n=15)：假手术组(Sham组)、缺血再灌注组(I/R组)和适度创伤疼痛6h后处理组(Tra-Post-6h组)。①Sham组：大鼠仅分离颈部血管,不进行大脑中动脉梗阻。②I/R组：采用MCAO模型建立大鼠局灶性脑缺血再灌注模型,缺血90min,再灌注24h。③Tra-Post-6h组：MCAO建模并缺血90min,大脑中动脉再灌注开始后的6小时在大鼠腹正中线作2cm的切口,皮肤至腹膜全层切开,之后分层缝合。三组大鼠在再灌注后观察24h,并在再灌注开始后的第6和24h对大鼠行改良的神经功能评分(modified neurological severity score, mNSS);用TTC法测定脑梗死容积；HE染色法观察脑组织形态；Nissl染色法计数存活神经细胞数目；免疫组化及Western Blot的方法观察及测定缺血侧大脑皮层半影区NCX1和NCX3蛋白的表达。 结果：①TTC染色：Sham组未见脑梗死；与I/R组比较,Tra-Post-6h组脑梗死容积显著缩小(P0.05)。②HE染色：Sham组大脑皮层神经元形态正常；与I/R组比较,Tra-Post-6h组脑缺血侧皮层半影区神经细胞层次较清楚,数目多且细胞形态较完整,间质水肿减轻；但是Tra-Post-6h组和I/R组脑缺血半影区细胞数目均较Sham组少。③Nissl染色：Sham组大脑皮层神经细胞数量正常；Tra-Post-6h组缺血半影区内存活的神经细胞数量比I/R组显著增加(P0.05),Tra-Post-6h和I/R组存活的神经细胞数量均较Sham组显著减少(P0.05)。④改良的神经功能评分：三组大鼠术前两次和术后两次评分在组间不存在显著差异(P0.05),各组大鼠的术前和术后两次评分组内比较也不存在统计学意义(P0.05)。⑤免疫组化及Western Blot结果：Tra-Post-6h组NCX1和NCX3蛋白在缺血侧大脑皮层半影区的表达均显著低于Sham组及I/R组(P0.05)；I/R组和Sham组之间两种蛋白的表达量不存在统计学差异(P0.05)。 结论：远隔创伤后处理可以对缺血再灌注的脑组织产生保护作用；这种脑保护作用可能通过减少NCX1和NCX3蛋白的表达来实现。
[Abstract]:Objective: to study the effects of (remote postconditioning of trauma, Tra-Post on cerebral ischemia / reperfusion injury in rats with focal cerebral ischemia-reperfusion injury and to study the sodium calcium exchanger (sodium-calcium exchangers,). The role of NCX in this post-processing. Methods: Forty-five male SD rats were randomly divided into three groups (n = 15): sham operation group (Sham group), ischemia reperfusion group (I / R group) and moderate traumatic pain treatment group (Tra-Post-6h group). Middle cerebral artery occlusion was not performed in 2I/R group: focal cerebral ischemia-reperfusion model was established with MCAO model in rats, ischemia 90 min, reperfusion 24h.3Tra-Post-6h group: MCAO modeling and ischemia 90 min. The middle cerebral artery (MCA) was cut into the abdominal median line of the rat at 6 hours after reperfusion. The skin was cut to the whole peritoneal layer, and then sutured in layers. The rats in the three groups were observed 24 hours after reperfusion, and the modified neurological function score (modified neurological severity score, mNSS);) was used to measure the volume of cerebral infarction by TTC method, and the morphology of brain tissue was observed by HE staining method at 6 and 24 hours after reperfusion. The number of surviving neurons was counted by Nissl staining and the expression of NCX1 and NCX3 in ischemic cerebral cortex penumbra was observed and measured by immunohistochemical and Western Blot methods. Results: 1TTC staining: no cerebral infarction was found in Sham group, and the volume of cerebral infarction was significantly reduced in Tra-Post-6h group compared with I / R group (P0.05). 2HE staining: the morphology of cortical neurons in Sham group was normal. Compared with the I / R group, the level of neurons in the cerebral ischemic penumbra of Tra-Post-6h group was clear, the number of neurons was more, the morphology of the cells was complete, and the interstitial edema was alleviated. However, the number of cerebral ischemic penumbra cells in Tra-Post-6h group and I / R group was less than that in Sham group. 3Nissl staining: the number of cortical neurons in Sham group was normal. The number of neurons surviving in ischemic penumbra in Tra-Post-6h group was significantly higher than that in I / R group (P0.05). The number of nerve cells survived in Tra-Post-6h and I / R groups was significantly lower than that in Sham group (P0.05). 4 improved neurological function score: there was no significant difference between the three groups in the scores of two times before and after operation (P0.05). There was no significant difference between the two scores before and after operation in each group (P0.05). 5 Immunohistochemical and Western Blot results: expression of NCX1 and NCX3 protein in ischemic cerebral cortex penumbra in Tra-Post-6h group It was significantly lower than that in Sham group and I / R group (P0.05). There was no significant difference in the expression of two proteins between I / R group and Sham group (P0.05). Conclusion: the post-traumatic treatment can protect the brain tissue from ischemia and reperfusion, which may be achieved by reducing the expression of NCX1 and NCX3 proteins.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743
本文编号：2372796
[Abstract]:Objective: to study the effects of (remote postconditioning of trauma, Tra-Post on cerebral ischemia / reperfusion injury in rats with focal cerebral ischemia-reperfusion injury and to study the sodium calcium exchanger (sodium-calcium exchangers,). The role of NCX in this post-processing. Methods: Forty-five male SD rats were randomly divided into three groups (n = 15): sham operation group (Sham group), ischemia reperfusion group (I / R group) and moderate traumatic pain treatment group (Tra-Post-6h group). Middle cerebral artery occlusion was not performed in 2I/R group: focal cerebral ischemia-reperfusion model was established with MCAO model in rats, ischemia 90 min, reperfusion 24h.3Tra-Post-6h group: MCAO modeling and ischemia 90 min. The middle cerebral artery (MCA) was cut into the abdominal median line of the rat at 6 hours after reperfusion. The skin was cut to the whole peritoneal layer, and then sutured in layers. The rats in the three groups were observed 24 hours after reperfusion, and the modified neurological function score (modified neurological severity score, mNSS);) was used to measure the volume of cerebral infarction by TTC method, and the morphology of brain tissue was observed by HE staining method at 6 and 24 hours after reperfusion. The number of surviving neurons was counted by Nissl staining and the expression of NCX1 and NCX3 in ischemic cerebral cortex penumbra was observed and measured by immunohistochemical and Western Blot methods. Results: 1TTC staining: no cerebral infarction was found in Sham group, and the volume of cerebral infarction was significantly reduced in Tra-Post-6h group compared with I / R group (P0.05). 2HE staining: the morphology of cortical neurons in Sham group was normal. Compared with the I / R group, the level of neurons in the cerebral ischemic penumbra of Tra-Post-6h group was clear, the number of neurons was more, the morphology of the cells was complete, and the interstitial edema was alleviated. However, the number of cerebral ischemic penumbra cells in Tra-Post-6h group and I / R group was less than that in Sham group. 3Nissl staining: the number of cortical neurons in Sham group was normal. The number of neurons surviving in ischemic penumbra in Tra-Post-6h group was significantly higher than that in I / R group (P0.05). The number of nerve cells survived in Tra-Post-6h and I / R groups was significantly lower than that in Sham group (P0.05). 4 improved neurological function score: there was no significant difference between the three groups in the scores of two times before and after operation (P0.05). There was no significant difference between the two scores before and after operation in each group (P0.05). 5 Immunohistochemical and Western Blot results: expression of NCX1 and NCX3 protein in ischemic cerebral cortex penumbra in Tra-Post-6h group It was significantly lower than that in Sham group and I / R group (P0.05). There was no significant difference in the expression of two proteins between I / R group and Sham group (P0.05). Conclusion: the post-traumatic treatment can protect the brain tissue from ischemia and reperfusion, which may be achieved by reducing the expression of NCX1 and NCX3 proteins.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743
【参考文献】
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1 Arunotai Siriussawakul;Ahmed Zaky;John D Lang;;Role of nitric oxide in hepatic ischemia-reperfusion injury[J];World Journal of Gastroenterology;2010年48期
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