脑缺血大鼠海马CA2区转铁蛋白受体及膜铁转运蛋白的表达及脑泰方提取物干预研究  

NTE treatment results in TFR and Fpn expression in hippocampus CA2 of rats subjected to cerebral ischemia

作　　者：廖君[1] 夏兴[1] 石咏梅[1] 易亚乔[1] 王国佐[1] 周瑜[1] 苏浩[1] 葛金文[1] 

机构地区：[1]湖南中医药大学,长沙410208

出　　处：《中华中医药杂志》2014年第5期1406-1411,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金青年项目(No.81303078);湖南省自然科学基金(No.12JJ6076);湖南省中医药管理局基金(No.201240);湖南省教育厅优秀青年项目基金(No.11B090);"中西医结合基础"湖南省"十二五"重点学科;"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;"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;中西医结合心脑疾病防治湖南省重点实验室;细胞生物学与分子技术湖南省高校重点实验室~~

摘　　要：目的:研究脑缺血大鼠海马CA2区转铁蛋白受体(TFR)和膜铁转运蛋白(Fpn)随时间表达,及益气活血中药脑泰方提取物(NTE)干预对TFR及Fpn表达的影响,阐明脑缺血后铁代谢失调导致神经元损伤的新机制及益气活血中药NTE治疗通过调节细胞内铁代谢达到抗脑缺血损伤的作用机制。方法:第一部分实验:随机将SD大鼠分为2h、6h、12h、24h、72h组,采用大脑中动脉栓塞法(MCAO)行大鼠局灶性脑缺血模型制备,分别在术后2、6、12、24、72h各时间点取海马,通过免疫组化及RT-PCR检测海马CA2区TFR、Fpn及TFR mRNA、FpnmRNA随时间表达。第二部分实验,随机将SD大鼠分为NTE低、中、高剂量组(3、9、27g/kg)、假手术组及模型组。各组大鼠预处理灌胃(ig)给药连续3d,每日1次,再行MCAO模型制备术,术后连续ig给药3d。术后第3天采用Zea Longa神经行为学评分标准记录大鼠的行为活动,尼氏染色观察海马CA2区神经元,免疫组化法及RT-PCR检测TFR、Fpn和TFR mRNA、Fpn mRNA表达。结果:12h、24h、72h组TFR表达明显增加(P<0.05),12h组Fpn的表达明显增加(P<0.05);NTE高剂量组神经行为学评分较模型组有显著降低(P<0.05);NTE高剂量组尼氏体多,胞核、胞仁较清晰显示;与模型组比较,NTE高剂量组TFR和TFR mRNA表达明显减少(P<0.05),与假手术组和模型组比较,NTE高剂量组Fpn和Fpn mRNA表达明显增高(P<0.05)。结论:铁调节失衡可能是导致脑缺血神经元损伤的新机制,脑缺血后NTE能通过干预大鼠海马CA2区TFR、Fpn的表达,调节神经元铁代谢,达到保护神经元,恢复神经功能的作用。Objective： To observe the expression of transferrin receptor （TFR） and ferroportin （Fpn） at different time points in rats after focal cerebral ischemia treated with or without traditional Chinese medicine NTE. Methods： The experiment was carded out by two steps： Firstly, SD rats were randomly divided into 2h, 6h, 12h, 24h, 72h groups after operation of middle cerebral artery occlusion （MCAO）, protein and mRNA level of TFR and Fpn were detected by immunohistochemistry and RT- PCR at above time points; Secondly, the rats were randomly allotted five groups as following： low dose group of NTE （3g/kg）, medial dose group of NTE （9g/kg）, high dose group of NTE （27g/kg）, sham operation group, mode/group. After three days of corresponding therapy by intragastric administration once a day, the regional cerebral ischemia model was reproduced by middle cerebral artery occlusion （MCAO） with suture method. Three days later, the rats treated by previous method. The third day, Neurological behavior of rats were detected by neurobehavioral testing the extent of neuronal damage were observed by Nissl staining. Protein and mRNA level of TFR and Fpn were detected by immunohistochemistry and RT-PCR at above treatment groups. Results： Twelve hours after the operation, expression of TFR significantly increased （P〈0.05）. Expression of Fpn in hippocampal CA2 reached the highest point at 12h after operation （P〈0.05）; Compared with model group, the high dose group of NTE （27g/kg） displayed a lower Neurological behavior score （P〈0.05）; Nissl body and nucleus were clear displayed in high dose group of NTE; High dose group of NTE showed lower level expression of TFR both at protein and mRNA level compared with model group （P〈0.05）, and high dose group of NTE showed higher lever expression of Fpn both at protein and mRNA level compared with sham operation group and model group （P〈0.05）. Conclusion： Imbalance of intracellular iron perhaps is the new mechanism of cerebral

关 键 词：转铁蛋白受体 膜铁转运蛋白 NTE提取物 脑缺血 

分 类 号：R285.5[医药卫生—中药学]