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目的比较正常及缺血损伤时大鼠血脑屏障上转运蛋白中P-糖蛋白(P-gp)时征表达及清脑宣窍方有效组分对其表达的影响。方法线栓法制备缺血性脑中风大鼠模型,按照不同再灌注时间,模型组再分为0、0.51、、26、、122、4 h,采用免疫组织化学法观察正常组与模型组大鼠脑皮质及海马缺血区P-gp的表达。除假手术组外,缺血动物于术后1 d,随机分为模型组、清脑宣窍方有效组分高、中、低剂量组、维拉帕米组,并于造模后1～5 d灌胃给予相应药物。末次给药后,处死动物,取材,检测大鼠脑皮质及海马区P-gp的表达。结果免疫组织化学染色结果显示,正常组与模型组大鼠脑皮质及海马缺血区均可见P-gp阳性染色。免疫组化半定量分析表明,与正常组比较,不同再灌注时间模型组P-gp表达量均有显著性差异(P<0.05),皮质及海马缺血区P-gp表达量均降低;与模型组比较,清脑宣窍方有效组分各剂量组及维拉帕米组对缺血性脑中风大鼠脑皮质及海马区P-gp表达量明显降低(P<0.05)。结论在缺血性脑中风病理状态下,大鼠脑血脑屏障上转运蛋白P-gp存在一定的时征表达,而且从一定程度上反映了特定病理状态下由于蛋白表达的改变而引起的血脑屏障通透性的变化。清脑宣窍方有效组分对缺血性脑中风大鼠脑皮质及海马缺血区P-gp表达具有显著的抑制作用。
Objective To compare the expression of P-glycoprotein (P-gp) on the blood-brain barrier (BBB) and the effect of effective components of Qingnao Xuanxiong Recipe on the expression of rat brain during normal and ischemic injury. Methods The ischemic stroke rat model was prepared by thread embolism. According to different reperfusion time, the model group was divided into 0, 0.51, 26, 122, 4 h. The normal group and the model group were observed by immunohistochemical method. Expression of P-gp in rat cerebral cortex and hippocampus ischemic area. In addition to the sham operation group, the ischemic animals were randomly divided into model group, high-, middle-, and low-dose groups of Qingnaoxuanfang group and verapamil group at the 1st day after operation. Five days after the administration of the corresponding drug. After the last administration, the animals were sacrificed and the material was taken to detect the expression of P-gp in the rat cerebral cortex and hippocampus. Results The results of immunohistochemical staining showed that P-gp positive staining was observed in the cerebral cortex and hippocampus in the normal and model groups. Semi-quantitative immunohistochemical analysis showed that compared with the normal group, there was a significant difference in P-gp expression levels in different reperfusion time model groups (P<0.05), and the P-gp expression levels in cortex and hippocampal ischemic region were decreased; Compared with the model group, the expression of P-gp in cerebral cortex and hippocampus of rats with ischemic cerebral apoplexy was significantly decreased in each dose group and Verapamil group of Qingnao Xuanxiong prescription (P<0.05). Conclusion In the pathological state of ischemic stroke, there is a certain transient expression of the transporter P-gp on the cerebral blood-brain barrier of rats, and to a certain extent, it reflects the blood caused by changes in protein expression under specific pathological conditions. Changes in brain barrier permeability. The effective components of Qingnao Xuanxiong Recipe have significant inhibitory effect on the expression of P-gp in cerebral cortex and ischemic hippocampus of rats with ischemic stroke.