中华神经科杂志
中華神經科雜誌
중화신경과잡지
Chinese Journal of Neurology
2013年
2期
91-94
,共4页
刘功禄%王开颜%郭辉%赵永波
劉功祿%王開顏%郭輝%趙永波
류공록%왕개안%곽휘%조영파
癫痫持续状态%内质网%应激%内切核糖核酸酶类%蛋白质丝氨酸苏氨酸激酶%JNK丝裂原活化蛋白激酶类%半胱氨酸天冬氨酸蛋白酶12
癲癇持續狀態%內質網%應激%內切覈糖覈痠酶類%蛋白質絲氨痠囌氨痠激酶%JNK絲裂原活化蛋白激酶類%半胱氨痠天鼕氨痠蛋白酶12
전간지속상태%내질망%응격%내절핵당핵산매류%단백질사안산소안산격매%JNK사렬원활화단백격매류%반광안산천동안산단백매12
Status epilepticus%Endoplasmic reticulum%Stress%Endoribonucleases%Protein-serine-threonine kinases%JNK mitogen-activated protein kinases%Caspase 12
目的 探讨肌醇酶1α(inositol requiring enzyme 1α,IRE 1α)介导的内质网应激相关凋亡分子在大鼠癫痫持续状态(SE)后海马神经元损伤中的作用.方法 Wistar大鼠按随机数字表随机分为对照组、SE组,SE组根据不同时间点又分为3、6、12、24、48 h组.免疫荧光法观察葡萄糖调节蛋白78(glucose-regulating protein 78 kd,GRP78)及磷酸化-IRE1α(p-IRE1α)在各组大鼠海马CA3区中的表达；蛋白质印迹检测磷酸化c-Jun N端激酶(p-JNK)、半胱氨酸天冬氨酸蛋白酶12(caspase 12)的表达变化；荧光TUNEL观察各组大鼠海马CA3区神经元凋亡变化.结果 免疫荧光结果显示,对照组仅见少量GRP78、p-IRE1α阳性神经元(分别为6.90％±0.96％,4.60％±1.12％),SE各亚组GRP78、p-IRE1α阳性神经元均增多,SE后12h达高峰(GRP78:87.45％±3.63％,F=356.82,P＜0.05；p-IRE1α:86.90％±3.82％,F=300.80,P＜0.05)；蛋白质印迹结果显示,与对照组相比,p-JNK、caspasel2在SE各亚组表达均增多,SE后12 h达高峰；同时TUNEL染色在SE各亚组均能检测到海马神经元凋亡,以SE后12h凋共最严重,与p-IRE1α、p-JNK、caspase12表达变化一致.结论 大鼠SE后诱发了内质网应激,表现为内质网应激标志分子GRP78表达增加；IRE1α可能通过磷酸化JNK和活化caspase12参与了SE后神经元凋亡损伤.
目的 探討肌醇酶1α(inositol requiring enzyme 1α,IRE 1α)介導的內質網應激相關凋亡分子在大鼠癲癇持續狀態(SE)後海馬神經元損傷中的作用.方法 Wistar大鼠按隨機數字錶隨機分為對照組、SE組,SE組根據不同時間點又分為3、6、12、24、48 h組.免疫熒光法觀察葡萄糖調節蛋白78(glucose-regulating protein 78 kd,GRP78)及燐痠化-IRE1α(p-IRE1α)在各組大鼠海馬CA3區中的錶達；蛋白質印跡檢測燐痠化c-Jun N耑激酶(p-JNK)、半胱氨痠天鼕氨痠蛋白酶12(caspase 12)的錶達變化；熒光TUNEL觀察各組大鼠海馬CA3區神經元凋亡變化.結果 免疫熒光結果顯示,對照組僅見少量GRP78、p-IRE1α暘性神經元(分彆為6.90％±0.96％,4.60％±1.12％),SE各亞組GRP78、p-IRE1α暘性神經元均增多,SE後12h達高峰(GRP78:87.45％±3.63％,F=356.82,P＜0.05；p-IRE1α:86.90％±3.82％,F=300.80,P＜0.05)；蛋白質印跡結果顯示,與對照組相比,p-JNK、caspasel2在SE各亞組錶達均增多,SE後12 h達高峰；同時TUNEL染色在SE各亞組均能檢測到海馬神經元凋亡,以SE後12h凋共最嚴重,與p-IRE1α、p-JNK、caspase12錶達變化一緻.結論 大鼠SE後誘髮瞭內質網應激,錶現為內質網應激標誌分子GRP78錶達增加；IRE1α可能通過燐痠化JNK和活化caspase12參與瞭SE後神經元凋亡損傷.
목적 탐토기순매1α(inositol requiring enzyme 1α,IRE 1α)개도적내질망응격상관조망분자재대서전간지속상태(SE)후해마신경원손상중적작용.방법 Wistar대서안수궤수자표수궤분위대조조、SE조,SE조근거불동시간점우분위3、6、12、24、48 h조.면역형광법관찰포도당조절단백78(glucose-regulating protein 78 kd,GRP78)급린산화-IRE1α(p-IRE1α)재각조대서해마CA3구중적표체；단백질인적검측린산화c-Jun N단격매(p-JNK)、반광안산천동안산단백매12(caspase 12)적표체변화；형광TUNEL관찰각조대서해마CA3구신경원조망변화.결과 면역형광결과현시,대조조부견소량GRP78、p-IRE1α양성신경원(분별위6.90％±0.96％,4.60％±1.12％),SE각아조GRP78、p-IRE1α양성신경원균증다,SE후12h체고봉(GRP78:87.45％±3.63％,F=356.82,P＜0.05；p-IRE1α:86.90％±3.82％,F=300.80,P＜0.05)；단백질인적결과현시,여대조조상비,p-JNK、caspasel2재SE각아조표체균증다,SE후12 h체고봉；동시TUNEL염색재SE각아조균능검측도해마신경원조망,이SE후12h조공최엄중,여p-IRE1α、p-JNK、caspase12표체변화일치.결론 대서SE후유발료내질망응격,표현위내질망응격표지분자GRP78표체증가；IRE1α가능통과린산화JNK화활화caspase12삼여료SE후신경원조망손상.
Objective To explore the role of inositol requiring enzyme 1 α (IRE1α) mediated endoplasmic reticulum stress associated apoptotic molecules in hippocampal neuronal injury in rats with status epilepsy following lithium-pilocarpine.Methods All 96 Wistar rats were randomly divided into control group and status epilepsy (SE) group.The SE group was further divided into 5 subgroups (3,6,12,24,48 h) according to different time points.pmmunofluorescence was used to observe the expressions of endoplasmic reticulum stress (ERS) markers glucose-regulating protein 78 kd (GRP78) and phosphoIRE1α (active form of endoplasmic reticulum resident protein IRE1α) at the CA3 area of rats in each group.Then,the expressions of IRElα mediated downstream apoptotie markers phospho-c-JunN-terminalkinase (JNK) and caspase12 were detected.Finally,TUNEL assay was used to observe neuronal apoptosis of hippocampal CA3 area at different time points after SE in rats.Results Immunofluorescence showed that GRP78 and phospho-IRE1α positive neurons were significantly increased in the SE subgroups compared with control group (6.90％ ± 0.96％,4.60％ ± 1.12％,respectively) and 12 h subgroup reached the peak (GRP78:87.45％ ±3.63％,F =356.82,P ＜0.05; phospho-IRE1α:86.90％ ±3.82％,F =300.80,P ＜ 0.05).Immunohistochemistry and Western blot demonstrated that the levels of phospho-JNK and caspase12 in the SE subgroups were significantly higher than that in the control group which reached the peak at 12 h after SE.The changes were in accord with phospho-IRE1α.Simultaneously,hippocampal neuronal apoptosis was detected in each SE subgroup and was most severe at 12 h after SE,which showed similar changes to the expressions of phospho-IRE1α,phospho-JNK and caspase12.Conclusions ERS was induced in rats following SE evidenced by increasing the expression of GRP78.IRE1α may promote hippocampal neuron apoptosis in rats following SE through activating JNK and caspase12.
