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
567-572
,共6页
张贵斌%付度关%王伟%徐运兰%王萍%田代实
張貴斌%付度關%王偉%徐運蘭%王萍%田代實
장귀빈%부도관%왕위%서운란%왕평%전대실
ERKs%U0126%CyclinD1%CyclinE%E2F%细胞周期%脑缺血
ERKs%U0126%CyclinD1%CyclinE%E2F%細胞週期%腦缺血
ERKs%U0126%CyclinD1%CyclinE%E2F%세포주기%뇌결혈
ERKs%U0126%CyclinD1%CyclinE%E2F%Cell cycle%Cerebral ischemia
目的 研究脑缺血后细胞外信号调节激酶(ERKs)对细胞周期调控的影响.方法 建立光化学法诱导大鼠局灶性脑缺血模型,分为脑缺血组(对照组及治疗组)和假手术组.治疗组于缺血前30min尾静脉注入U0126溶液,对照组尾静脉注入相同体积不含U0126的DMSO稀释溶液.应用免疫组织荧光化学法观察细胞周期蛋白D1(CyclinD1)和细胞周期蛋白E(CyclinE)阳性细胞表达:免疫印迹(Weaem blot)检测磷酸化ERK1/2(pERK1/2)、CyclinD1和CyclinE的蛋白表达;半定量逆转录-聚合酶链反应(PT-PCR)观察转录因子E2F mRNA的表达.结果 治疗组CyclinD1和CyclinE阳性表达的细胞数较对照组显著减少(P<0.05);缺血对照组pERK1/2蛋白表达显著强于治疗组(P<0.05),4h时间点表达最为明显,12h时间点恢复到基线水平,CyclinD1和CyelinE表达6h开始升高,12h表达最为明显,治疗组较对照组显著减弱(P<0.05);缺血对照组E2F mRNA的表达显著强于治疗组和假手术组(P<0.05),以7d表达最为明显.结论 ERKs在大鼠脑缺血中发挥重要作用,抑制脑缺血引起的ERK1/2磷酸化,可降低细胞周期蛋白CyelinD1、CyclinE和E2F的表达.即ERKs可影响细胞周期的调控.
目的 研究腦缺血後細胞外信號調節激酶(ERKs)對細胞週期調控的影響.方法 建立光化學法誘導大鼠跼竈性腦缺血模型,分為腦缺血組(對照組及治療組)和假手術組.治療組于缺血前30min尾靜脈註入U0126溶液,對照組尾靜脈註入相同體積不含U0126的DMSO稀釋溶液.應用免疫組織熒光化學法觀察細胞週期蛋白D1(CyclinD1)和細胞週期蛋白E(CyclinE)暘性細胞錶達:免疫印跡(Weaem blot)檢測燐痠化ERK1/2(pERK1/2)、CyclinD1和CyclinE的蛋白錶達;半定量逆轉錄-聚閤酶鏈反應(PT-PCR)觀察轉錄因子E2F mRNA的錶達.結果 治療組CyclinD1和CyclinE暘性錶達的細胞數較對照組顯著減少(P<0.05);缺血對照組pERK1/2蛋白錶達顯著彊于治療組(P<0.05),4h時間點錶達最為明顯,12h時間點恢複到基線水平,CyclinD1和CyelinE錶達6h開始升高,12h錶達最為明顯,治療組較對照組顯著減弱(P<0.05);缺血對照組E2F mRNA的錶達顯著彊于治療組和假手術組(P<0.05),以7d錶達最為明顯.結論 ERKs在大鼠腦缺血中髮揮重要作用,抑製腦缺血引起的ERK1/2燐痠化,可降低細胞週期蛋白CyelinD1、CyclinE和E2F的錶達.即ERKs可影響細胞週期的調控.
목적 연구뇌결혈후세포외신호조절격매(ERKs)대세포주기조공적영향.방법 건립광화학법유도대서국조성뇌결혈모형,분위뇌결혈조(대조조급치료조)화가수술조.치료조우결혈전30min미정맥주입U0126용액,대조조미정맥주입상동체적불함U0126적DMSO희석용액.응용면역조직형광화학법관찰세포주기단백D1(CyclinD1)화세포주기단백E(CyclinE)양성세포표체:면역인적(Weaem blot)검측린산화ERK1/2(pERK1/2)、CyclinD1화CyclinE적단백표체;반정량역전록-취합매련반응(PT-PCR)관찰전록인자E2F mRNA적표체.결과 치료조CyclinD1화CyclinE양성표체적세포수교대조조현저감소(P<0.05);결혈대조조pERK1/2단백표체현저강우치료조(P<0.05),4h시간점표체최위명현,12h시간점회복도기선수평,CyclinD1화CyelinE표체6h개시승고,12h표체최위명현,치료조교대조조현저감약(P<0.05);결혈대조조E2F mRNA적표체현저강우치료조화가수술조(P<0.05),이7d표체최위명현.결론 ERKs재대서뇌결혈중발휘중요작용,억제뇌결혈인기적ERK1/2린산화,가강저세포주기단백CyelinD1、CyclinE화E2F적표체.즉ERKs가영향세포주기적조공.
Objective To investigate the effect of extracellular signal-regulated kinases (ERKs) on cell cycle regulation after ischemia. Methods Ischemic model was induced by photochemistry method. Animals were divided randomly into cerebral ischemia groups (control and treatment groups) and sham group. Rats in treatment group were subjected to U0126 solution injection at 30 rain pre-ischemia through caudal veins, and animals in control group were subjected to identical volume DMSO solution without U0126. Positive immunostaining for CyclinD1 and CyclinE were detected by immunohistofluorescence method. Expressions of phosphorylated ERK1/2 (pERK1/2), CyclinD1, and CyclinE proteins were examined by Western blot in ischemic slide of brain cortex. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of transcription factor E2F mRNA in ischemic slide of brain cortex. Results The numbers of CyclinD1 and CyclinE positive cells were highly decreased in the U0126-treated group (P<0.05 vs vehicle-treated group). Expression of pERK1/2 protein in the ischemic group was significantly higher than that in the U0126-treated group, which peaked at 4h, and decreased to the baseline at 12 after ischemia. While the expression levels of CyclinD1 and CyclinE in the U0126-treated group were increased at 6h post injury, peaked at 12 after injury (P<0.05 vs that in vehicle-treated group). In addition, expression of E2F mRNA in the vehicle-treated group was significantly higher than those in the sham-operated group and the U0126-treated group (P<0.05). Conclusions ERK pathway plays a very important role in cerebral ischemia. Inhibiting ERK1/2 phosphorylation post-ischemia reduces the expressions of CyclinD1, CyclinE and E2F, which indicates that ERK can affect cell cycle regulation.
