NHE1和GLUTs蛋白水平在缺血性脑损伤再灌注大鼠脑内的表达意义

Expression Significance of NHE1 and Gluts Protein Levels in the Brain of Rats with Ischemic Brain Injury Reperfusion

探讨线栓法构建大脑中动脉缺血(MCAO)大鼠模型脑组织Na^(+)/H^(+)交换蛋白1型(Na^(+)/H^(+)exchanger protein 1,NHE1)和GLUTs蛋白的表达水平。方法为:50只雄性SD大鼠随机分为假手术组(6只)、MCAO模型组(44只),通过线栓法构建大鼠大脑中动脉缺血(Middle cerebral artery occlusion,MCAO)再灌注模型,造模成功后的2 h,4 h,6 h,12 h,24 h各处死6只MCAO大鼠,免疫组化方法观察脑组织形态结构的改变,采用免疫荧光法和western-blot法检测大鼠左侧脑内NHE1,GLUT1及GLUT3表达,并与假手术组比较。与假手术组比较,MCAO模型组大鼠出现明显的神经功能障碍,病理检测结果显示脑组织出现明显的梗死灶,脑组织含水量明显升高,NHE1蛋白和GLUT1蛋白表达水平明显升高(P<0.01),GLUT3蛋白表达明显减少,显著低于假手术组(P<0.01)。结果显示,脑缺氧缺血后脑内NHE1和GLUT3蛋白表达升高,GLUT3蛋白的表达降低可导致脑能量代谢降低,加重缺氧缺血后神经元的损伤。

To investigate the expression level of Na^(+)/H^(+)exchange protein 1(NHE1)and Gluts protein in the brain tissue of MCAO model constructed by thread thrombolysis.Methods:Fifty male SD rats were randomly divided into sham operation group(6 rats)and MCAO model group(44 rats).The Middle cerebral artery occlusion(MCAO)reperfusion model was established by suture method.After successful modeling,2h,4h,6h,12h,After 24 hours,6 MCAO rats were sacrificed,and the morphological and structural changes of brain tissue were observed by immunohistochemistry.The expressions of NHE1,GLUT1 and GLUT3 in the left brain of rats were detected by immunofluorescence and Western-blot,and compared with those in the sham operation group.Results:Compared with the sham operation group,the rats in the MCAO model group had obvious neurological dysfunction.Pathological examination results showed that the brain tissue had obvious infarction,the water content in the brain tissue was significantly increased,and the protein expression levels of NHE1 and GLUT1 were significantly increased(P<0.01).GLUT3 protein expression was significantly decreased compared with sham operation group(P<0.01).Conclusion:The expression of NHE1 and GLUT3 protein in brain increases after cerebral hypoxia ischemia,and the decreased expression of GLUT3 protein can lead to decreased brain energy metabolism and aggravate neuronal injury after cerebral hypoxia ischemia.