中国中西医结合外科杂志
中國中西醫結閤外科雜誌
중국중서의결합외과잡지
CHINESE JOURNAL OF SURGERY OF INTEGRATED TRADITIONAL AND WESTERN MEDICINE
2015年
3期
274-278
,共5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%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%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%감성성섬유세포생장인자%린산기순3격매-단백격매B%린산화세포외신호조절격매1/2
Propofol%cerebral ischemia/reperfusion injury%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(CI/RI)中的作用及可能机制。方法：采用氧糖剥夺再灌注（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组和丙泊酚+OGD/RP联合组。采用MTT法检测皮质神经元存活率，Annexin V-PI检测细胞凋亡情况，RT-PCR方法检测碱性成纤维细胞生长因子（bFGF）的mRNA表达，Western blot检测丙泊酚对皮质神经元内bFGF、磷酸化蛋白激酶B（pPKB）和磷酸化细胞外信号调节激酶1/2（pERK1/2）表达。采用小干扰RNA构建bFGF沉默的细胞。结果：丙泊酚能够显著促进CI/RI后神经元的存活，抑制其凋亡，OGD/RP处理组神经元凋亡率为43.2%，以10 mg/L的丙泊酚预处理后细胞的凋亡率即降为19.5%（P＜0.05）。与对照组（1.02±0.03）相比较，OGD处理后细胞中bFGF的含量（0.78±0.06）显著下调（P＜0.05），丙泊酚处理的皮质神经元中bFGF含量(1.43±0.04)显著高于OGD处理组（P＜0.05）。沉默的bFGF或者施加蛋白激酶B(PI3K-pPKB）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- pPKB以及ERK1/2的激活。结论：丙泊酚可以通过上调bFGF的表达，激活PI3K- pPKB和ERK1/2的信号通路，减轻体外培养神经元凋亡/再灌注损伤，从而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(CI/RI)中的作用及可能機製。方法：採用氧糖剝奪再灌註（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組和丙泊酚+OGD/RP聯閤組。採用MTT法檢測皮質神經元存活率，Annexin V-PI檢測細胞凋亡情況，RT-PCR方法檢測堿性成纖維細胞生長因子（bFGF）的mRNA錶達，Western blot檢測丙泊酚對皮質神經元內bFGF、燐痠化蛋白激酶B（pPKB）和燐痠化細胞外信號調節激酶1/2（pERK1/2）錶達。採用小榦擾RNA構建bFGF沉默的細胞。結果：丙泊酚能夠顯著促進CI/RI後神經元的存活，抑製其凋亡，OGD/RP處理組神經元凋亡率為43.2%，以10 mg/L的丙泊酚預處理後細胞的凋亡率即降為19.5%（P＜0.05）。與對照組（1.02±0.03）相比較，OGD處理後細胞中bFGF的含量（0.78±0.06）顯著下調（P＜0.05），丙泊酚處理的皮質神經元中bFGF含量(1.43±0.04)顯著高于OGD處理組（P＜0.05）。沉默的bFGF或者施加蛋白激酶B(PI3K-pPKB）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- pPKB以及ERK1/2的激活。結論：丙泊酚可以通過上調bFGF的錶達，激活PI3K- pPKB和ERK1/2的信號通路，減輕體外培養神經元凋亡/再灌註損傷，從而增加皮質神經元的存活。
목적：탐토병박분재뇌결혈/재관주손상(CI/RI)중적작용급가능궤제。방법：채용양당박탈재관주（OGD/RP）법체외구건결혈/재관주세포모형，장세포분위대조조、OGD/RP조화병박분+OGD/RP연합조。채용MTT법검측피질신경원존활솔，Annexin V-PI검측세포조망정황，RT-PCR방법검측감성성섬유세포생장인자（bFGF）적mRNA표체，Western blot검측병박분대피질신경원내bFGF、린산화단백격매B（pPKB）화린산화세포외신호조절격매1/2（pERK1/2）표체。채용소간우RNA구건bFGF침묵적세포。결과：병박분능구현저촉진CI/RI후신경원적존활，억제기조망，OGD/RP처리조신경원조망솔위43.2%，이10 mg/L적병박분예처리후세포적조망솔즉강위19.5%（P＜0.05）。여대조조（1.02±0.03）상비교，OGD처리후세포중bFGF적함량（0.78±0.06）현저하조（P＜0.05），병박분처리적피질신경원중bFGF함량(1.43±0.04)현저고우OGD처리조（P＜0.05）。침묵적bFGF혹자시가단백격매B(PI3K-pPKB）이급pERK1/2신호통로억제제도회도치세포존활솔현저하강（P＜0.05），억제PI3K- pPKB이급ERK1/2적격활。결론：병박분가이통과상조bFGF적표체，격활PI3K- pPKB화ERK1/2적신호통로，감경체외배양신경원조망/재관주손상，종이증가피질신경원적존활。
Objective To study the effect of propofol on cerebral ischemia/reperfusion injury(CI/RI) and the relative mechanism. Methods The cerebral ischemia/reperfusion model was produced by oxygen glucose deprivation and reperfusion in vitro. Cells were grouped into control group, OGD/RP group and propofol+OGD/RPgroup. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the proliferative effect of propofol. Apoptot?ic 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 (pPKB) and phosphorylated extracellular signal-regulated kinase 1/2(pERK1/2) protein expression were detected by <br> Western blotting, silence of bFGF in cortical neurons by small interfering RNA. Results Propofol could signif?icantly promote the viability of neurons and inhibit cell apoptosis after ischemia reperfusion injury. The apopto?sis 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 were significantly lower than those in 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 pPKB and pERK1/2, and activate the two signaling pathways. Silence the expression of bFGF or treatment with the in?hibitor of phosphotylinosital 3 kinase-protein kinase B (PI3K-pPKB) and pERK1/2 signal pathway both resulted in the decrease of neurons viability (P<0.05), and the inhibition of PI3K-pPKB and pERK1/2 activation. Conclusion Propofol can activate PI3K-pPKB and pERK1/2 signal pathway via upregulating the expression of bFGF, and promote the survival of cortical neurons.
