华中科技大学学报(医学版)
華中科技大學學報(醫學版)
화중과기대학학보(의학판)
ACTA UNIVERSITATIS MEDICINAE TONGJI
2014年
2期
168-172
,共5页
邓镇%罗淼珊%赵元淑%余涵%谢柳%陈丽%朱晓琴%胡景鑫%雷水生
鄧鎮%囉淼珊%趙元淑%餘涵%謝柳%陳麗%硃曉琴%鬍景鑫%雷水生
산진%라묘산%조원숙%여함%사류%진려%주효금%호경흠%뢰수생
NMDA受体NR2A亚单位%侧脑室注射%P-糖蛋白%癫痫持续状态
NMDA受體NR2A亞單位%側腦室註射%P-糖蛋白%癲癇持續狀態
NMDA수체NR2A아단위%측뇌실주사%P-당단백%전간지속상태
NR2A subunit of N-methyl-D-aspartate receptor%intracerebroventricular injection%P-glycoprotein%sta-tus epilepticus
目的:探讨谷氨酸N-甲基-D-天冬氨酸受体(NMDA)的NR2A亚单位(GluN2A)选择性抑制剂PEAQX对癫痫持续状态大鼠脑内P-糖蛋白(P-glycoprotein ,P-gp)表达的影响。方法将实验动物随机分为3组:对照组(先侧脑室注射生理盐水,再腹腔注射生理盐水)、致痫组(先侧脑室注射生理盐水,再腹腔注射戊四氮致痫)和抑制剂干预组(先侧脑室注射PEAQX ,再腹腔注射戊四氮致痫),观察各组大鼠行为学表现;使用BL-420E生物机能实验系统记录各组大鼠脑电的改变;采用RT-qPCR和Western blot技术,检测各组大鼠海马组织中P-gp mRNA及蛋白表达的变化。结果行为学观察显示:对照组无明显癫痫发作,致痫组癫痫发作快且明显,诱导癫痫持续状态成功,而抑制剂干预组癫痫发作潜伏期长且不明显;脑电结果显示:对照组无明显痫样波,致痫组记录到棘波、尖波、棘-慢复合波等痫样波,抑制剂干预组痫样波减弱或消失;RT-qPCR结果显示:致痫组海马中 P-gp mRNA表达增加,抑制剂干预组 P-gp mRNA表达明显减少;Western blot结果显示:致痫组海马中P-gp蛋白表达增强,抑制剂干预组P-gp表达明显减弱。结论 GluN2A选择性抑制剂抑制了癫痫的发作,并抑制了P-gp的表达,其机制可能与GluN2A选择性抑制剂抑制了GluN2A的活性有关,并提示GluN2A选择性抑制剂可能与减少癫痫耐药的产生有关。
目的:探討穀氨痠N-甲基-D-天鼕氨痠受體(NMDA)的NR2A亞單位(GluN2A)選擇性抑製劑PEAQX對癲癇持續狀態大鼠腦內P-糖蛋白(P-glycoprotein ,P-gp)錶達的影響。方法將實驗動物隨機分為3組:對照組(先側腦室註射生理鹽水,再腹腔註射生理鹽水)、緻癇組(先側腦室註射生理鹽水,再腹腔註射戊四氮緻癇)和抑製劑榦預組(先側腦室註射PEAQX ,再腹腔註射戊四氮緻癇),觀察各組大鼠行為學錶現;使用BL-420E生物機能實驗繫統記錄各組大鼠腦電的改變;採用RT-qPCR和Western blot技術,檢測各組大鼠海馬組織中P-gp mRNA及蛋白錶達的變化。結果行為學觀察顯示:對照組無明顯癲癇髮作,緻癇組癲癇髮作快且明顯,誘導癲癇持續狀態成功,而抑製劑榦預組癲癇髮作潛伏期長且不明顯;腦電結果顯示:對照組無明顯癇樣波,緻癇組記錄到棘波、尖波、棘-慢複閤波等癇樣波,抑製劑榦預組癇樣波減弱或消失;RT-qPCR結果顯示:緻癇組海馬中 P-gp mRNA錶達增加,抑製劑榦預組 P-gp mRNA錶達明顯減少;Western blot結果顯示:緻癇組海馬中P-gp蛋白錶達增彊,抑製劑榦預組P-gp錶達明顯減弱。結論 GluN2A選擇性抑製劑抑製瞭癲癇的髮作,併抑製瞭P-gp的錶達,其機製可能與GluN2A選擇性抑製劑抑製瞭GluN2A的活性有關,併提示GluN2A選擇性抑製劑可能與減少癲癇耐藥的產生有關。
목적:탐토곡안산N-갑기-D-천동안산수체(NMDA)적NR2A아단위(GluN2A)선택성억제제PEAQX대전간지속상태대서뇌내P-당단백(P-glycoprotein ,P-gp)표체적영향。방법장실험동물수궤분위3조:대조조(선측뇌실주사생리염수,재복강주사생리염수)、치간조(선측뇌실주사생리염수,재복강주사무사담치간)화억제제간예조(선측뇌실주사PEAQX ,재복강주사무사담치간),관찰각조대서행위학표현;사용BL-420E생물궤능실험계통기록각조대서뇌전적개변;채용RT-qPCR화Western blot기술,검측각조대서해마조직중P-gp mRNA급단백표체적변화。결과행위학관찰현시:대조조무명현전간발작,치간조전간발작쾌차명현,유도전간지속상태성공,이억제제간예조전간발작잠복기장차불명현;뇌전결과현시:대조조무명현간양파,치간조기록도극파、첨파、극-만복합파등간양파,억제제간예조간양파감약혹소실;RT-qPCR결과현시:치간조해마중 P-gp mRNA표체증가,억제제간예조 P-gp mRNA표체명현감소;Western blot결과현시:치간조해마중P-gp단백표체증강,억제제간예조P-gp표체명현감약。결론 GluN2A선택성억제제억제료전간적발작,병억제료P-gp적표체,기궤제가능여GluN2A선택성억제제억제료GluN2A적활성유관,병제시GluN2A선택성억제제가능여감소전간내약적산생유관。
Objective To investigate the effect of the selective inhibitor of glutamate N-methyl-D-aspartate receptor (NM-DA) NR2A subunit (GluN2A) PEAQX on the P-glycoprotein (P-gp) expression in the brain of rats with status epileptic-us.Methods The experimental animals were randomly divided into three groups :control group ,in which the rats first under-went intracerebroventricular injection of saline and then intraperitoneal injection of saline ;epileptic group ,in which the rats were first intracerebroventricularly injected with saline and then intraperitoneally injected with pentylenetetrazole ;inhibitor interven-tion group ,in which the rats were intracerebroventricularly administrated with PEAQX ,and then intraperitoneally with pentyle-netetrazole.The behavior manifestations of rats were observed ,and the electroencephalogram (EEG) changes of rats were recor-ded with BL-420E biological and functional experimental system.The expression changes of P-gp in the hippocampus in each group were examined by real-time quantitative PCR (RT-qPCR) and Western blot.Results Behavioral observation showed that rats in the control group had no obvious epileptic seizures ,and those in epileptic group presented with significant and fast sei-zures ,which suggested that status epilepticus models were successfully established.In the inhibitor intervention group ,a long incubation period of seizures was found and the seizures were less obvious.EEG showed no significant epileptiform waves in the control group but vertex sharp transient wave ,sharp wave and composite wave and so on in the epileptic group.In the inhibitor intervention group ,the waves diminished or disappeared.The results of RT-qPCR showed that the expression of P-gp mRNA in the hippocampus of epileptic group was increased obviously ,while it was significantly reduced in the inhibitor intervention group.The results of Western blot were consistent with those of RT-qPCR.Conclusion The GluN2A inhibitor PEAQX could reduce the epileptic seizures and the expression of P-gp by inhibiting the activity of GluN2A.The findings suggested that the se-lective inhibitors of GluN2A might attenuate the drug resistance in epilepsy.