国际麻醉学与复苏杂志
國際痳醉學與複囌雜誌
국제마취학여복소잡지
INTERNATIONAL JOURNAL OF ANESTHESIOLOGY AND RESUSCITATION
2015年
3期
198-203
,共6页
周国强%巩晓洁%傅蕊%周晓东
週國彊%鞏曉潔%傅蕊%週曉東
주국강%공효길%부예%주효동
丙泊酚%脑缺血/再灌注损伤%氧糖剥夺%碱性成纤维细胞生长因子%磷酸肌醇3激酶-蛋白激酶B%磷酸化细胞外信号调节激酶1/2
丙泊酚%腦缺血/再灌註損傷%氧糖剝奪%堿性成纖維細胞生長因子%燐痠肌醇3激酶-蛋白激酶B%燐痠化細胞外信號調節激酶1/2
병박분%뇌결혈/재관주손상%양당박탈%감성성섬유세포생장인자%린산기순3격매-단백격매B%린산화세포외신호조절격매1/2
Propofol%Cerebral ischemia/reperfusion injury%Oxygen glucose deprivation%Basic fibroblast growth factor%Phosphotylinosital 3 kinase-protein kinase B%Phosphorylated extracellular signal-regulated kinase 1/2
目的 探讨丙泊酚在脑缺血/再灌注损伤(ischemia/reperfusion injury,I/RI)中发挥的作用及其具体机制. 方法 采用氧糖剥夺再灌注(oxygen-glucose deprivation/reperfusion,OGD/RP)法体外构建缺血/再灌注细胞模型,将细胞分为对照组、OGD/RP组、丙泊酚+OGD/RP组.采用甲基噻唑基四唑(methyl thiazolyl tetrazolium,MTT)法检测皮质神经细胞存活率,Annexin V-PI检测细胞凋亡情况,即时聚合酶链式反应(real-time polymerase chain reaction,RT-PCR)方法检测碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)的mRNA表达情况,免疫印迹法(Western blot)检测丙泊酚对皮质神经细胞内bFGF,磷酸化蛋白激酶B(phosphorylated protein kinase B,pAkt)以及磷酸化细胞外信号调节激酶1/2 (phosphorylated extr acellular signal-regulated kinase 1/2,pERK 1/2)蛋白表达的影响;采用小干扰RNA构建bFGF沉默的细胞. 结果 OGD/RP处理组神经细胞凋亡率为43.2％,经10 mg/L的丙泊酚预处理后,细胞的凋亡率降为19.5％.与对照组比较,OGD处理后,细胞中bFGF的含量显著下调(P＜0.05),丙泊酚处理的皮质神经元中bFGF含量显著高于OGD处理组(P＜0.05).丙泊酚能够上调pAKT以及pERK1/2的表达,激活这两条信号通路.沉默bFGF或者施加磷酸肌醇3激酶-蛋白激酶B(phosphotylinosital 3 kinase-protein kinase B,PI3K-Akt)以及pERK1/2信号通路抑制剂都会导致细胞存活率显著下降(P＜0.05),抑制PI3K-Akt以及pERK1/2的激活. 结论 丙白酚可以通过上调bFGF的表达,激活PI3K-Akt和ERK 1/2信号通路,增加皮质神经元的存活.
目的 探討丙泊酚在腦缺血/再灌註損傷(ischemia/reperfusion injury,I/RI)中髮揮的作用及其具體機製. 方法 採用氧糖剝奪再灌註(oxygen-glucose deprivation/reperfusion,OGD/RP)法體外構建缺血/再灌註細胞模型,將細胞分為對照組、OGD/RP組、丙泊酚+OGD/RP組.採用甲基噻唑基四唑(methyl thiazolyl tetrazolium,MTT)法檢測皮質神經細胞存活率,Annexin V-PI檢測細胞凋亡情況,即時聚閤酶鏈式反應(real-time polymerase chain reaction,RT-PCR)方法檢測堿性成纖維細胞生長因子(basic fibroblast growth factor,bFGF)的mRNA錶達情況,免疫印跡法(Western blot)檢測丙泊酚對皮質神經細胞內bFGF,燐痠化蛋白激酶B(phosphorylated protein kinase B,pAkt)以及燐痠化細胞外信號調節激酶1/2 (phosphorylated extr acellular signal-regulated kinase 1/2,pERK 1/2)蛋白錶達的影響;採用小榦擾RNA構建bFGF沉默的細胞. 結果 OGD/RP處理組神經細胞凋亡率為43.2％,經10 mg/L的丙泊酚預處理後,細胞的凋亡率降為19.5％.與對照組比較,OGD處理後,細胞中bFGF的含量顯著下調(P＜0.05),丙泊酚處理的皮質神經元中bFGF含量顯著高于OGD處理組(P＜0.05).丙泊酚能夠上調pAKT以及pERK1/2的錶達,激活這兩條信號通路.沉默bFGF或者施加燐痠肌醇3激酶-蛋白激酶B(phosphotylinosital 3 kinase-protein kinase B,PI3K-Akt)以及pERK1/2信號通路抑製劑都會導緻細胞存活率顯著下降(P＜0.05),抑製PI3K-Akt以及pERK1/2的激活. 結論 丙白酚可以通過上調bFGF的錶達,激活PI3K-Akt和ERK 1/2信號通路,增加皮質神經元的存活.
목적 탐토병박분재뇌결혈/재관주손상(ischemia/reperfusion injury,I/RI)중발휘적작용급기구체궤제. 방법 채용양당박탈재관주(oxygen-glucose deprivation/reperfusion,OGD/RP)법체외구건결혈/재관주세포모형,장세포분위대조조、OGD/RP조、병박분+OGD/RP조.채용갑기새서기사서(methyl thiazolyl tetrazolium,MTT)법검측피질신경세포존활솔,Annexin V-PI검측세포조망정황,즉시취합매련식반응(real-time polymerase chain reaction,RT-PCR)방법검측감성성섬유세포생장인자(basic fibroblast growth factor,bFGF)적mRNA표체정황,면역인적법(Western blot)검측병박분대피질신경세포내bFGF,린산화단백격매B(phosphorylated protein kinase B,pAkt)이급린산화세포외신호조절격매1/2 (phosphorylated extr acellular signal-regulated kinase 1/2,pERK 1/2)단백표체적영향;채용소간우RNA구건bFGF침묵적세포. 결과 OGD/RP처리조신경세포조망솔위43.2％,경10 mg/L적병박분예처리후,세포적조망솔강위19.5％.여대조조비교,OGD처리후,세포중bFGF적함량현저하조(P＜0.05),병박분처리적피질신경원중bFGF함량현저고우OGD처리조(P＜0.05).병박분능구상조pAKT이급pERK1/2적표체,격활저량조신호통로.침묵bFGF혹자시가린산기순3격매-단백격매B(phosphotylinosital 3 kinase-protein kinase B,PI3K-Akt)이급pERK1/2신호통로억제제도회도치세포존활솔현저하강(P＜0.05),억제PI3K-Akt이급pERK1/2적격활. 결론 병백분가이통과상조bFGF적표체,격활PI3K-Akt화ERK 1/2신호통로,증가피질신경원적존활.
Objective To study the effect of propofol on cerebral ischemia/reperfusion injury (I/RI) and the relative mechanism.Methods The cerebral ischemia/reperfusion model was produced by oxygen glucose deprivation and reperfusion in vitro,methyl thiazolyl tetrazolium (MTT) assay was used to detect the proliferative effect of propofol.Apoptotic cells were detected with Annexin V staining.The expression level of basic fibroblast growth factor (bFGF) mRNA was measured by RT-PCR.The expression levels of bFGF,phosphorylated protein kinase B (pAkt) and phosphorylated extracellular signal-regulated kinase 1/2 (pERK1/2) protein expression were detected by Western blot.Silence of bFGF in cortical neurons by small interfering RNA.Results The apoptosis rate of neurons in oxygen-glucose deprivation/reperfusion (OGD/RP) group was 43.2％.After treated with 10 mg/L propofol,the apoptosis rate of neurons was reduced to 19.5％.The expression levels of bFGF protein in OGD treated group was significantly lower than the control group(P＜0.05),and the bFGF level was markedly upregulated in propofol treated group(P＜0.05).Propofol could upregulate the expression of pAkt and pERK1/2,and activate the two signaling pathways.Silence the expression of bFGF or treatment with the inhibitor of phosphotylinosital 3 kinase-protein kinase B (PI3K-Akt) and ERK1/2 signal pathway both resulted in the decrease of neurons viability (P＜0.05),and the inhibition of PI3K-Akt and ERK1/2 activation.Conclusions Propofol could activate PI3K-Akt and ERK1/2 signal pathway via upregulating the expression of bFGF,and promote the survival of cortical neurons.
