低氧诱导因子-1α与马桑内酯点燃SD大鼠难治性颞叶癫痫耐药模型耐药性的相关性研究  

Correlational study of hypoxia-inducible factor-1 and pharmacoresistance of Sprague-Dawley rat temporal lobe epilepsy model kindled by coriaria lactone

作　　者：陈蕾[1] 李耀华[1] 曾天芳[1] 周东[1] 

机构地区：[1]四川大学华西医院神经内科,成都610041

出　　处：《解放军医学杂志》2012年第5期415-420,共6页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(30900470/C090301)~~

摘　　要：目的探讨难治性颞叶癫痫中低氧诱导因子-1α（HIF-1α）在不同脑区表达水平的变化及其与耐药蛋白MDR1/Pgp表达的关系。方法以SD大鼠作为研究对象,设置点燃组和对照组。点燃组处理方法：以马桑内酯（CL） 0.4ml/kg肌注,1次/72h,在给药1~5次期间判定点燃模型是否构建成功,选择符合标准的大鼠继续给药,1次/6d,共30次;最后点燃组共存留7只大鼠。对照组5只大鼠,在同时间点给予0.4ml/kg生理盐水肌注。取大鼠脑组织标本,制备后采用实时荧光定量PCR（RQ-PCR）、免疫组织化学（IHC）和Western blotting三种方法分别检测HIF-1α、MDR1/Pgp在脑组织中的表达情况。结果 IHC显示,HIF-1α在点燃组主要表达于大鼠海马、颞叶皮质等部位的神经元和神经胶质细胞,在对照组仅见微弱表达。Pgp的分布及表达情况与HIF-1α一致。RQ-PCR显示,点燃组HIF-1α mRNA在海马中的表达为对照组的1.741倍,在颞叶中的表达是对照组的1.395倍。Pgp mRNA在海马及颞叶的表达分别为对照组的2.297倍和1.474倍。Western blotting显示,点燃组海马HIF-1α蛋白表达（1.284±0.166）较对照组（0.763±0.117）明显增高（P〈0.05）,且点燃组颞叶皮质中HIF-1α蛋白（1.350±0.105）亦较对照组（0.941±0.078）明显增高（P〈0.05）。Pgp的表达与HIF-1α表达呈一致变化,即点燃组海马和颞叶中Pgp的表达（0.831±0.086,0.919±0.129）较对照组（0.441±0.053,0.643±0.095）明显增高（P〈0.05）。另外,点燃组Pgp蛋白和mRNA在海马中的表达较颞叶高（P〈0.05）。结论 HIF-1α在大鼠颞叶癫痫耐药模型的海马和颞叶皮质中的表达较正常大鼠高,且与Pgp表达的空间分布一致。HIF-1α与Pgp表达的相关性提示HIF-1α可能与难治性颞叶癫痫的耐药性相关。Objective To investigate changes in the expression of hypoxia-inducible factor-1（HIF-1） in different brain regions in a pharmacoresistance model of temporal lobe epilepsy,and explore its relation to the expression of drug resistance gene/ protein multidrug-resistance-gene1（MDR1）/P-glycoprotein （Pgp）. Methods A kindling model of pharmacoresistant temporal lobe epilepsy was developed by injecting Sprague-Dawley （SD） rats with coriaria lactone （CL）. Normal SD rats were injected with normal sodium（NS） served as control group.The expressions of HIF-1,MDR1/Pgp in brain tissues were detected by realtime fluorescence-based quantitative polymerase chain reaction （RQ-PCR）, immunohistochemistry （IHC）, Western blotting, respectively.Results IHC showed that the expressions of HIF-1 in neurons and glial cells were higher in the hippocampus and temporal cortex than in other brain regions in kindled rats,and the regional distribution of HIF-1 was similar to that of Pgp.RQ-PCR and Western blotting showed that the expressions of HIF-1 in the hippocampus and temporal cortex were higher in kindled group than in control group（P0.05）,and the expressions of MDR1/Pgp in gene and protein levels were consistent with the alterations of HIF-1 in both groups.Conclusion HIF-1 expressions in hippocampus and temporal cortex in kindled rats are higher than those in control rats,and it corresponds to the regional distribution of Pgp.The correlation of HIF-1 and MDR1/Pgp expressions indicates that HIF-1 may be related to the drug resistance of refractory temporal lobe epilepsy.

关 键 词：低氧诱导因子-1 基因 MDR P糖蛋白 难治性癫痫 SPRAGUE-DAWLEY大鼠 马桑内酯 0577. 7402(2012)05-.0415-.06 

分 类 号：R742.11[医药卫生—神经病学与精神病学]