中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2008年
6期
549-553
,共5页
张树华%朱长庚%刘庆莹%魏瑛%王伟
張樹華%硃長庚%劉慶瑩%魏瑛%王偉
장수화%주장경%류경형%위영%왕위
星形胶质细胞%条件培养液%马桑内酯%谷氨酸%受体
星形膠質細胞%條件培養液%馬桑內酯%穀氨痠%受體
성형효질세포%조건배양액%마상내지%곡안산%수체
Astrocyte%Conditionedmedium%Coriarialactone%Glutamate%Receptor
目的 揭示马桑内酯(CL)激活的星形胶质细胞(Ast)条件培养液(ACM)对大鼠脑内谷氨酸(Glu)及其受体GluR2表达的影响.方法 取成年健康雄性SD大鼠48只,采用随机数字表法分为对照组(16只)和CL组(32只),对照组侧脑室注射未加任何刺激物的ACM 10μL,CL组侧脑室注射CL激活的ACM 10μL;按注射后取材时间不同又分为2h、4h、8h和12h四个亚组,对照组每亚组4只,CL组每亚组8只.观察两组大鼠的行为表现,用免疫组化、免疫荧光检测脑内Glu和GluR2表达的变化,Western blot检测脑内GluR2含量的变化.结果 CL组大鼠有痫样发作,而对照组无痫样发作;免疫组化和免疫荧光检测结果显示,CL组皮质和海马区Glu表达较对照组显著增强,4h时差异有统计学意义(P<0.05),而CL组皮质和海马区GluR2的表达较对照组弱,4h时差异有统计学意义(P<0.05).Western blot结果显示,CL组4个时间点的GluR2表达均较对照组含量显著降低,差异有统计学意义(P<0.05).结论 CL激活的ACM能显著增强脑内神经元Glu的表达,降低GIuR2的表达,进而诱发癫痫.
目的 揭示馬桑內酯(CL)激活的星形膠質細胞(Ast)條件培養液(ACM)對大鼠腦內穀氨痠(Glu)及其受體GluR2錶達的影響.方法 取成年健康雄性SD大鼠48隻,採用隨機數字錶法分為對照組(16隻)和CL組(32隻),對照組側腦室註射未加任何刺激物的ACM 10μL,CL組側腦室註射CL激活的ACM 10μL;按註射後取材時間不同又分為2h、4h、8h和12h四箇亞組,對照組每亞組4隻,CL組每亞組8隻.觀察兩組大鼠的行為錶現,用免疫組化、免疫熒光檢測腦內Glu和GluR2錶達的變化,Western blot檢測腦內GluR2含量的變化.結果 CL組大鼠有癇樣髮作,而對照組無癇樣髮作;免疫組化和免疫熒光檢測結果顯示,CL組皮質和海馬區Glu錶達較對照組顯著增彊,4h時差異有統計學意義(P<0.05),而CL組皮質和海馬區GluR2的錶達較對照組弱,4h時差異有統計學意義(P<0.05).Western blot結果顯示,CL組4箇時間點的GluR2錶達均較對照組含量顯著降低,差異有統計學意義(P<0.05).結論 CL激活的ACM能顯著增彊腦內神經元Glu的錶達,降低GIuR2的錶達,進而誘髮癲癇.
목적 게시마상내지(CL)격활적성형효질세포(Ast)조건배양액(ACM)대대서뇌내곡안산(Glu)급기수체GluR2표체적영향.방법 취성년건강웅성SD대서48지,채용수궤수자표법분위대조조(16지)화CL조(32지),대조조측뇌실주사미가임하자격물적ACM 10μL,CL조측뇌실주사CL격활적ACM 10μL;안주사후취재시간불동우분위2h、4h、8h화12h사개아조,대조조매아조4지,CL조매아조8지.관찰량조대서적행위표현,용면역조화、면역형광검측뇌내Glu화GluR2표체적변화,Western blot검측뇌내GluR2함량적변화.결과 CL조대서유간양발작,이대조조무간양발작;면역조화화면역형광검측결과현시,CL조피질화해마구Glu표체교대조조현저증강,4h시차이유통계학의의(P<0.05),이CL조피질화해마구GluR2적표체교대조조약,4h시차이유통계학의의(P<0.05).Western blot결과현시,CL조4개시간점적GluR2표체균교대조조함량현저강저,차이유통계학의의(P<0.05).결론 CL격활적ACM능현저증강뇌내신경원Glu적표체,강저GIuR2적표체,진이유발전간.
Objective To explore the effects of coriafia lactone (CL)-activated astrocytes (Ast) conditioned medium (ACM) on the expressions of glutamate (Glu) and GluR2 in the brain of rat. Methods Asts of hippocampus were cultured according to the McCarthy and DeVellis's method, and then the ACM was collected. Forty-eight male adult Sprague-Dawley (SD) rats were randomly divided into the control group (n=16) and the CL group (n=32). Rats in the control group were administered 10 μL ACM I. C. V., which was not added any stimulating substance. Rats of the CL group were injected I. C. V. 10 μL CL-activated ACM. The rats in both groups were subdivided into post-injection 2,4,8,12h subgroups, 4 in each subgroup in the control group and 8 in each subgroup in the CL group. The behaviors of the rats were observed and the expressions of Glu and GluR2 in the cerebral cortex and hippocampus were detected with immunohistochemistry and immunofluorescence. The content of GluR2 was tested with Western blot. Results The rats injected with CL-activated ACM showed seizure activities, whereas the rats of the control group showed no seizure activities. The expression of Glu in cerebral cortex and hippocampus in the brains injected with CL-activated ACM was increased compared with the control group at 4h (P<0.05), but the expression of GluR2 was attenuated compared with the control group at 4h(P<0.05). The results of GluR2 in the cerebral cortex and hippocampus detected with Western blot were different significantly with control group (P<0.05). Conclusion CL-activated ACM can enhance the expression of Glu and reduce the expression of GluR2 in the brain of rat, resulting in the activation of AMPA pathway and the Ca2+ influx, and then induce seizure activities.
