丙泊酚通过PI3K/AKT/HIF-1α通路减轻氧糖剥夺-复糖复氧大鼠海马神经元损伤的研究  

Propofol alleviates hippocampal neuronal damage in oxygen glucose deprivation reoxygenation rats via PI3K/AKT/HIF-1αpathway

作　　者：王鹏涛 斯坎带尔·吾守尔 黄玮 李宗滨 邓文强 张劬 WANG Peng-tao;SIKANDAIER·Wushouer;HUANG Wei;LI Zong-bin;DENG Wen-qiang;ZHANG Qian(Department of Anesthesiology,Third People’s Hospital of Xinjiang Uygur Autonomous Region,Urümqi 830099,Xinjiang,China;Department of Anesthesiology,Fifth Affiliated Hospital of Xinjiang Medical University,Urümqi830054,Xinjiang,China)

机构地区：[1]新疆维吾尔自治区第三人民医院麻醉科,新疆乌鲁木齐830099 [2]新疆医科大学第五附属医院麻醉科,新疆乌鲁木齐830054

出　　处：《生物医学工程与临床》2025年第1期1-6,共6页Biomedical Engineering and Clinical Medicine

摘　　要：目的探讨丙泊酚通过磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)/低氧诱导因子-1α(HIF-1α)通路减轻氧糖剥夺-复糖复氧大鼠海马神经元损伤。方法采用孕16 d左右Wistar雌性大鼠3只(平均体质量220 g),取海马组织并原代培养海马神经元7 d。然后将海马神经元随机分为对照组、模型组、丙泊酚组和PI3K过表达组,除对照组,其他3组均复制氧糖剥夺-复糖复氧模型,丙泊酚组给予丙泊酚1.2μg/mL,PI3K过表达组在原代神经元培养第3天进行PI3K基因过表达转染,第7天复制模型并给予丙泊酚1.2μg/mL。各组继续培养24 h,采用流式细胞术检测神经元凋亡率,实时荧光定量聚合酶链式反应(qRT-PCR)检测PI3K、AKT和HIF-1αmRNA表达,Western blot法检测PI3K、p-PI3K、AKT、p-AKT和HIF-1α蛋白表达,酶联免疫吸附分析(ELISA)法检测细胞培养液中肿瘤坏死因子(TNF)-α和白细胞介素(IL)-6浓度,ELISA检测还原性谷胱甘肽(GSH)和超氧化物歧化酶(SOD)活性。结果与对照组相比[(4.5±0.5)%,1.03±0.06、1.06±0.08、1.09±0.09,0.35±0.04、0.34±0.03、0.27±0.05,(56.4±6.8)pg/mL、(35.2±4.5)pg/mL,(21.2±4.3)μmol/g、(32.6±7.8)U/mg],模型组神经元凋亡率[(58.4±5.7)%]和PI3K、AKT、HIF-1αmRNA表达量(1.99±0.12、2.11±0.14、2.56±0.18)及p-PI3K、p-AKT和HIF-1α蛋白表达量(0.57±0.03、0.62±0.04、0.53±0.02)、TNF-α和IL-6浓度[(203.6±45.2)pg/mL、(152.3±23.6)pg/mL]明显增加,而GSH和SOD活性[(7.8±1.3)μmol/g、(10.1±3.2)U/mg]降低(t=265.84、25.65、105.23、34.03、10.12、4.03、6.47、7.93、3.45、3.02、3.65,P<0.05)。与模型组相比,丙泊酚组神经元凋亡率[(31.5±4.3)%]、PI3K、AKT和HIF-1αmRNA表达量(1.36±0.09、1.39±0.11、1.98±0.13)及p-PI3K、p-AKT和HIF-1α蛋白表达量(0.45±0.05、0.46±0.04、0.39±0.05)、TNF-α和IL-6浓度[(154.2±32.3)pg/mL、(102.1±15.9)pg/mL]明显下降,而GSH和SOD活性[(14.4±3.2)μmol/g、(23.2±6.6)U/mg]升高(t=8.96、8.96、5.33、10.02、16.44、3.04、3.1Objective To investigate the effect of propofol on alleviating hippocampal neuron damage in rats with oxygen glucose deprivation and reoxygenation via phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/hypoxic inducer factor-1α(HIF-1α)pathway.Methods Three female Wistar rats at about 16 days of gestation(mean body mass 220 g)were sacrificed.The hippocampal neurons were taken and primary hippocampal neurons were cultured for 7 days,and hippocampal neurons were randomly divided into control group,model group,propofol group and PI3K over-expression group.Except for control group,the other three groups replicated oxygen glucose deprivation reoxygenation model.The propofol group was given propofol 1.2μg/mL,the PI3K over-expression group was transfected with PI3K gene on the 3rd day of primary neuron culture,and model was replicated on the 7th day and propofol 1.2μg/mL was given.Each group was cultured for 24 hours,the apoptosis rate of neurons was detected by flow cytometry,the expression of PI3K,AKT and HIF-1αmRNA was detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).The expression of PI3K,p-PI3K,AKT,p-AKT and HIF-1αprotein was detected by Western blot.The concentrations of tumor necrosis factor(TNF)-αand interleukin(IL)-6 in cell culture medium were detected by enzyme-linked immunosorbent assay(ELISA).The activities of reduced glutathione(GSH)and superoxide dismutase(SOD)were detected by ELISA.Results Compared with control group[(4.5±0.5)%,1.03±0.06,1.06±0.08,1.09±0.09,0.35±0.04,0.34±0.03,0.27±0.05,(56.4±6.8)pg/mL,(35.2±4.5)pg/mL,(21.2±4.3)μmol/g,(32.6±7.8)U/mg],neuronal apoptosis rate[(58.4±5.7)%],mRNA expression levels of PI3K,AKT and HIF-1αmRNA(1.99±0.12,2.11±0.14,2.56±0.18),protein expression levels of p-PI3K,p-AKT and HIF-1α(0.57±0.03,0.62±0.04,0.53±0.02),concentrations of TNF-αand IL-6[(203.6±45.2)pg/mL,(152.3±23.6)pg/mL]were significantly increased,while activities of GSH and SOD[(7.8±1.3)μmol/g,(10.1±3.2)U/mg]were decreased in model gr

关 键 词：丙泊酚 氧糖剥夺-复糖复氧 海马神经元 磷脂酰肌醇3-激酶 蛋白激酶B 低氧诱导因子-1Α 

分 类 号：R614[医药卫生—麻醉学]