P-糖蛋白在大鼠脑缺血再灌注后的表达变化

发布时间：2018-04-05 20:57

本文选题：P-糖蛋白　切入点：脑缺血　出处：《重庆医学》2017年32期

【摘要】：目的观察大鼠局灶性脑缺血90min再灌注后P-糖蛋白(P-gp)在脑组织中的表达及其变化规律。方法采用线栓法建立大鼠脑缺血再灌注模型,选取26只成年健康SD雄性大鼠,其中1只用于2,3,5-三苯基氯化四氮唑(TTC)染色,其余25只随机分为对照组(n=5)、假手术组(n=5)、脑缺血再灌注1、3、7d组(n=5)。用免疫组织化学DAB单标,观察P-gp在正常脑组织和缺血脑组织中的分布变化;用Mdr-1抗体分别和神经元标记物(Neun)、星形胶质细胞标记物(GFAP)、微血管内皮细胞标记物(CD31)进行免疫荧光双标,观察P-gp在缺血再灌注后脑组织的表达;用实时定量PCR技术检测脑缺血再灌注后大脑皮质及纹状体微血管P-gp mRNA的表达变化。结果对照组仅在脑组织微血管内皮细胞表达P-gp,缺血再灌注组除微血管内皮细胞表达P-gp外,在部分神经元及星形胶质细胞的突起也有表达。脑缺血再灌注后1d皮质P-gp mRNA表达降低,但3d表达明显增高,7d又下降,其升高和降低水平与对照组及假手术组比较,差异有统计学意义(P0.05)。脑缺血再灌注后1、3、7d纹状体P-gp mRNA表达均增多,其中以1、3d增多明显,与对照组和假手术组比较,差异有统计学意义(P0.05)。结论脑缺血再灌注后P-gp在大脑皮层及纹状体微血管P-gp的表达呈现不同的趋势,这可能是脑组织的自我保护机制之一。
[Abstract]:Objective to observe the expression and changes of P-gp in rat brain after focal cerebral ischemia (90min) reperfusion.Methods A rat model of cerebral ischemia-reperfusion was established by the method of thread embolization. 26 adult male SD rats were selected, one of them was used for TTCstaining of 2-titriphenyl 5-triphenyl tetrazolium chloride (TTC).The other 25 rats were randomly divided into control group (n = 5), sham operation group (n = 5), cerebral ischemia reperfusion group (n = 5) and cerebral ischemia reperfusion group (n = 5).Immunohistochemical DAB was used to observe the distribution of P-gp in normal brain tissue and ischemic brain tissue, and immunofluorescence double labeling was performed with Mdr-1 antibody and neuronal marker, astrocyte marker, microvascular endothelial cell marker (CD31), respectively.To observe the expression of P-gp in cerebral tissue after ischemia-reperfusion, and to detect the expression of P-gp mRNA in cerebral cortex and striatum by real-time quantitative PCR.Results in the control group, P-gp was expressed only in the microvascular endothelial cells of the brain, and in the ischemic reperfusion group, the expression of P-gp was also found in some neurons and astrocytes in addition to the expression of P-gp in the microvascular endothelial cells.The expression of P-gp mRNA decreased at 1 day after cerebral ischemia and reperfusion, but increased significantly at day 3 and decreased again on day 7. There was a significant difference in the level of P-gp expression between the control group and the sham operation group (P 0.05).The expression of P-gp mRNA in striatum was increased on the 7th day after cerebral ischemia-reperfusion, especially on the 1st day after reperfusion. The difference was statistically significant compared with the control group and sham-operation group (P 0.05).Conclusion the expression of microvascular P-gp in cerebral cortex and striatum is different after cerebral ischemia reperfusion, which may be one of the self-protective mechanisms of brain tissue.
【作者单位】：昆明医科大学基础医学院人体解剖学与组织胚胎学系;昆明医科大学科研中心;昆明医科大学高等教育研究所;
【基金】：国家自然科学基金资助项目(81060101) 云南省应用基础研究计划项目(2013FZ056) 昆明医科大学青年教师培养特殊支持计划项目(J1301306612)
【分类号】：R743

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