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目的研究沙鼠脑缺血再灌注后脑内磷酸化的Tau蛋白(PHF-1)和高尔基体表面蛋白标记抗体(58K-9)表达的变化及美满霉素干预对它们表达的影响。方法采用夹闭双侧颈总动脉的方法制作沙鼠脑缺血再灌注模型;50只沙鼠随机分为正常对照组、假手术组、缺血再灌注组、美满霉素干预组(美满霉素干预+缺血再灌注);美满霉素干预组的沙鼠在实验前1d开始腹腔注射美满霉素(剂量为45mg/kg,Bid);缺血再灌注组、美满霉素干预组分别在缺血再灌注(IR)后6h、1d、3d、7d的各时间点分批处死动物,快速取脑组织制作标本,采用免疫组织化学方法检测各组沙鼠脑内PHF-1、58K-9的表达水平。结果(1)正常对照组与假手术组沙鼠脑组织中可见少量PHF-1的阳性表达,2组比较无差别(P>0.05),且阳性信号主要位于神经元和胶质细胞胞浆中;缺血10min分别再灌注3d、7d后脑组织中PHF-1表达增加,7d组表达高于3d组;3、7d组与正常对照组及假手术组比较差异有显著性(P<0.01);(2)正常对照组与假手术组沙鼠脑组织中可见大量58K-9的阳性表达,2组比较无差别(P>0.05),且阳性信号主要位于神经元和胶质细胞胞浆中;缺血10min分别再灌注3、7d后脑组织中58K-9表达减少,7d组表达低于3d组;3、7d组与正常对照组及假手术组比较差异有显著性(P<0.01);(3)美满霉素干预组在缺血10min再灌注6h、1、3、7d时沙鼠脑组织中PHF-1、58K-9表达趋势同缺血再灌注组,但3、7d组PHF-1阳性率明显低于缺血再灌注组,58K-9阳性率高于缺血再灌注组,相同时间点比较差异有显著性(P<0.01)。结论(1)沙鼠脑缺血再灌注后脑组织PHF-1表达增高,提示Tau蛋白发生高度磷酸化改变;58K-9表达减少,提示高尔基体结构与功能的损坏;(2)美满霉素干预可使沙鼠脑缺血再灌注后脑组织PHF-1表达下调、使58K-9表达上调,美满霉素有神经保护作用。
Objective To investigate the changes of phosphorylated Tau protein (PHF-1) and Golgi surface marker protein (58K-9) in gerbils after cerebral ischemia-reperfusion and the effect of minocycline on their expression. Methods Fifty gerbils were randomly divided into normal control group, sham operation group, ischemia-reperfusion group and minocycline intervention group (+ 45mg / kg, Bid). The rats in ischemia-reperfusion group and minocycline-treated group received intraperitoneal injection of minocycline at the dose of 45mg / kg, The rats were sacrificed at 6h, 1d, 3d and 7d after ischemia-reperfusion (IR), and the brain tissues were quickly harvested to make specimens. Immunohistochemistry was used to detect the changes of PHF-1, 58K-9 The level of expression. Results (1) A small amount of PHF-1 positive expression was found in the brain tissue of normal and sham operation group, there was no difference between the two groups (P> 0.05), and the positive signal mainly located in the cytoplasm of neurons and glial cells (P <0.01). The expression of PHF-1 in brain tissue increased 7 days after reperfusion for 10 minutes and reached the level of 7th day after reperfusion for 3 days and 3 days respectively. There was significant difference between 3 and 7 days group and normal control group and sham operation group (P <0.01). (2) A large number of 58K-9 positive cells were found in the brain tissue of gerbils in normal control group and sham operation group, there was no difference between the two groups (P> 0.05), and the positive signals mainly located in the cytoplasm of neurons and glial cells; The expression of 58K-9 in brain tissue decreased at 3 and 7 days after reperfusion 10 min after ischemia, and the expression of 58K-9 in 7d group was lower than that of 3d group at 3 and 7 days after reperfusion (P <0.01) 3) In the minocycline-treated group, the expression of PHF-1, 58K-9 in the brain tissue of the gerbils was similar to that of the ischemia-reperfusion group at 6h, 1,3,7d after ischemia for 10min, but the PHF-1 The positive rate of 58K-9 was significantly lower than that of ischemia-reperfusion group (P <0.01). Conclusions (1) The expression of PHF-1 in brain tissue of gerbils after cerebral ischemia-reperfusion increased, suggesting a hyperphosphorylation of Tau protein, a decrease of 58K-9 expression, suggesting the damage of Golgi structure and function. (2) It can down-regulate the expression of PHF-1 in brain tissue of gerbils after cerebral ischemia and reperfusion, and up-regulate the expression of 58K-9. Neuromycin has a neuroprotective effect.