灯盏花素对小鼠局灶性脑缺血损伤的保护作用及与脑组织P-糖蛋白表达研究  

Studys of protective effects of breviscapine on acute cerebral ischemia injury in mice and brain tissue P-gp mRNA and protein expression relationship

作　　者：金桂兰[1,2] 覃慧林 石孟琼[4] 王爱玲[3] 程凡[3] 徐帮[1,2] 曾晓[1,2] 

机构地区：[1]三峡大学人民医院 [2]宜昌市第一人民医院 [3]三峡大学生物与制药学院 [4]三峡大学医学院,宜昌443002

出　　处：《中药药理与临床》2015年第1期76-79,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：湖北省自然科学基金项目(No.2012FFB03705)

摘　　要：目的:研究灯盏花素对小鼠局灶性脑缺血损伤的保护作用及与脑组织中P-糖蛋白表达关系。方法:雄性6周龄昆明小鼠,采用完全随机设计分组法分为5组:假手术组、模型组、灯盏花素低、高剂量(5,10mg/kg)和尼莫地平组(4.0mg/kg),除假手术组和模型组灌胃给予蒸馏水20ml/kg外,其余各组灌胃给予相应剂量的药物。7天后制作大鼠大脑中动脉闭塞模型,缺血24小时后,观察进行神经症状评分、斜板试验,取大脑并用2,3,5-三苯基四氮唑染色后测定梗塞梗死面积和用失重法计算脑组织含水量;用实时定量荧光PCR和蛋白质印迹法检测脑组织中P-糖蛋白的基因和蛋白表达。结果:灯盏花素(5,10mg/kg)和尼莫地平能显著改善局灶性脑缺血损伤后小鼠的神经症状,降低脑梗死面积和脑含水量为(68.3±5.1,66.1±4.8和65.3±5.7);降低脑组织中P-糖蛋白的基因为(0.68±0.11,0.61±0.10和0.73±0.08)和蛋白表达为(0.39±0.05,0.34±0.06和0.50±0.09),与模型组比较,具有显著性差异,其中以灯盏花素10mg/kg剂量组作用效果更为显著。结论:灯盏花素对小鼠局灶性脑缺血损伤具有较好的保护作用,其作用机制可能与抑制P-糖蛋白的表达水平,升高脑内灯盏花素的药物浓度有关。Objective： To investigate the protective effects of breviscapine on acute cerebral ischemia injury in mice and brain tissue P-glycoprotein（ P-gp） mRNA and protein expression relationship. Methods： Mice were divided into low dose（ 5mg/kg） and high dose（ 10. 0mg/kg） breviscapine groups,nimodipine group and sham group respectively,after pretreating homologous drugs for 7days. The mouse model of middle cerebral artery occlusion was induced,and neurological symptom scoring and inclined board testing were performed with in 24 h after I / R.The infarct volume and neuron density of mice were recorded. Real-time PCR and western blotting were applied to detect the P-gp mRNA and protein expressions in brain tissue. Results： Breviscapine mightsignificantly reduced the neurological deficits,infarct size and water content of acute cerebral ischemia injury in mice for 0. 39 ± 0. 05,0. 34 ± 0. 06,0. 50 ± 0. 09,decrease the mRNA level for 0. 68 ± 0. 11,0. 61 ± 0.10,0. 73 ± 0. 08 and protein level for 0. 39 ± 0. 05,0. 34 ± 0. 06,0. 50 ± 0. 09 of P-gp in brain tissue,respectively. There was a significant difference compared with the model group（ P 〈0. 05 or P 〈0. 01）,and with the dose of breviscapine progressively increasing,all of these changes became much stronger. Conclusion： The results suggest that breviscapine has preferably protective effects on acute cerebral ischemia injury in mice,and the potential mechanism of action was to suppress the the expression of P-gp,increase the drug concentration in brain of breviscapine.

关 键 词：灯盏花素 局灶性脑缺血 P-糖蛋白 

分 类 号：R285.5[医药卫生—中药学]