摘要
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(b FGF)的m RNA表达,Western blot检测丙泊酚对皮质神经元内b FGF、磷酸化蛋白激酶B(p PKB)和磷酸化细胞外信号调节激酶1/2(p ERK1/2)表达。采用小干扰RNA构建b FGF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b FGF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b FGF含量(1.43±0.04)显著高于OGD处理组(P<0.05)。沉默的b FGF或者施加蛋白激酶B(PI3K-p PKB)以及p ERK1/2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-p PKB以及ERK1/2的激活。结论:丙泊酚可以通过上调b FGF的表达,激活PI3K-p PKB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。
Objective To study the effect of propofol on cerebral ischemia/reperfusion injury（CI/RI） andthe relative mechanism.MethodsThe cerebral ischemia/reperfusion model was produced by oxygen glucosedeprivation 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（b FGF）mRNA was measured by RT-PCR. The expression levels of bFGF, phosphorylated protein kinase B（pPKB） and phosphorylated extracellular signal-regulated kinase1/2（pERK1/2） protein expression were detected by Western blotting, silence of b FGF in cortical neurons by small interfering RNA.ResultsPropofol 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 10mg/L propofol, the apoptosis rate of neurons was reduced to 19.5%. The expression levels of b FGF protein inOGD treated group were significantly lower than those in the control group（P 0.05）, and the b FGF level wasmarkedly upregulated in propofol treated group（P 0.05）. Propofol could upregulate the expression of p PKBand p ERK1/2, and activate the two signaling pathways. Silence the expression of b FGF or treatment with the in-hibitor of phosphotylinosital 3 kinase-protein kinase B（PI3K-p PKB） and p ERK1/2 signal pathway both resultedin the decrease of neurons viability（P 0.05）, and the inhibition of PI3K-p PKB and p ERK1/2 activation.ConclusionPropofol can activate PI3K-p PKB and p ERK1/2 signal pathway via upregulating the expressionof bFGF, and promote the survival of cortical neurons.
出处
《中国中西医结合外科杂志》
CAS
2015年第3期274-278,共5页
Chinese Journal of Surgery of Integrated Traditional and Western Medicine
