大鼠脑缺血后海马CA2区FLVCR和BCRP的表达及益气活血法干预  

Research on Expression of FLVCR and BCRP in Hippocamp CA2 of Rats Subjected to Cerebral Ischemia and Traditional Chinese Medicine of Supplementing Qi and Activating Blood Circulation Intervention Effects

作　　者：廖君[1] 陈懿[1] 夏兴[1] 罗琳[1] 石咏梅[1] 余清平[1] 葛金文[1] 

机构地区：[1]湖南中医药大学,湖南省长沙市410208

出　　处：《中国动脉硬化杂志》2014年第11期1091-1096,共6页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金青年项目(81303078);湖南省自然科学基金(12JJ6076);湖南省自然科学基金(13JJ3100);湖南省中医药管理局基金(201240);湖南省教育厅项目基金(12C0269);"中西医结合基础"湖南省"十二五"重点学科资助;"中医药防治心脑血管疾病基础研究"湖南省自然科学创新群体基金资助;"中西医结合防治心脑血管疾病的相关基础研究"湖南省高校科技创新团队资助

摘　　要：目的研究脑缺血后血红素铁转运蛋白人类猫白血病C亚类病毒受体(feline leukemia virus subgroup C receptor,FLVCR)及胸腺癌抵抗蛋白(breast cancer resistance protein,BCRP)随时间表达及益气活血法脑泰方(黄芪、川芎、地龙、僵蚕)提取物干预对FLVCR及BCRP表达的影响。方法实验分两步进行,第一部分实验:随机将SD大鼠分为2 h、6 h、12 h、24 h、72 h组,采用大脑中动脉栓塞法(middle cerebral artery occlusion,MCAO)行大鼠局灶性脑缺血模型制备,分别在术后2 h、6 h、12 h、24 h、72 h各时间点取材,通过免疫组织化学及RT-PCR检测FLVCR和BCRP及FLVCR mRNA和BCRP mRNA随时间表达的变化。第二部分实验,随机将SD大鼠分为假手术组、模型组、脑泰方低、中、高剂量组(3、9、27 g/kg)。各组大鼠预处理藻胃给药连续3天,MCAO模型制备术后连续藻胃给药3天,每日1次。术后3天取材,电镜检测海马神经元内铁样颗粒分布,免疫组织化学及RT-PCR检测FLVCR和BCRP及FLVCR mRNA和BCRP mRNA表达。结果 72 h组FLVCR表达明显降低(P<0.05),BCRP的表达2 h、6h较高,12 h、24 h、72 h表达明显降低(P<0.05);电镜下脑泰方各剂量组神经元内铁样颗粒聚集较模型组减少。脑泰方高剂量组FLVCR及FLVCR mRNA表达明显增加(P<0.05),BCRP表达无明显改变。结论大鼠脑缺血后第72 h FLVCR表达下降,益气活血中药脑泰方提取物干预可增加脑缺血后FLVCR的表达,促进血红素铁的外排,可能是治疗脑缺血损伤的新机制。Aim To observe the expression of FLVCR and BCRP at different time points and effects of traditional herbal medicines NTE（ extract of Huang Qi,Chuan Xiong,Di Long,Jiang Can） on FLVCR and BCRP after Focal Cerebral Ischemia in rats. Methods The experiment is carried out by two steps. Firstly,SD rats were randomly divided into 2 h,6 h,12 h,24 h,72 h groups,after operation of middle cerebral artery occlusion（ MCAO）,FLVCR,BCRP and FLVCR-mRNA,BCRP-mRNA expressions were detected by immunohistochemistry and RT-PCR just at 2 h,6 h,12 h,24 h,72 h time points.Secondly,The rats were randomly allotted 5 groups as following ： sham operation group,model group,low dose group of NTE（ 3g / kg）,medial dose group of NTE（ 9 g / kg）,large dose group of NTE（ 27 g / kg）. After 3 days of corresponding therapy by intragastric administration once every day,the regional cerebral ischemia model was reproduced by middle cerebral artery occlusion（ MCAO） with suture method. Following 3 days,the rats were treated with previous method. On the third day,The iron particles of neurons were observed by Electron microscopic. FLVCR and BCRP in hippocampal CA2 region were measured by immunohistochemical method and FLVCR-mRNA and BCRP-mRNA were detected by RT-PCR with marketable kit. Results Expression of FLVCR in hippocamp CA2 descend at the point of 72 nd hour after operation（ P 〈 0. 05）,and expression of BCRP decreased at 12 h after operation,and at 24 h and 72 h maintained low level（ P 〈 0. 05）; each dose group of NTE showed less iron particles of neuron in the electron microscope scanning; expression of the FLVCR and FLVCR-mRNA increased in each NTE dose group（ P 〈 0. 05）,the expression of BCRP had no obvious change. Conclusion Expression of FLVCR in hippocamp CA2 descend at the point of 72 hour after operation,the large dose group of NTE can increase the expression of FLVCR and FLVCR-mRNA after cerebral ischemia. Increased expression of FLVCR and excessive iron efflux,may be the mechanism of Yiqihuoxue Nao

关 键 词：猫白血病C亚类病毒受体 胸腺癌抵抗蛋白 脑泰方提取物 脑缺血 

分 类 号：R255[医药卫生—中医内科学]