咪达唑仑减轻缺氧/复氧诱导的小鼠海马神经元损伤  

Midazolam attenuates hypoxia-reoxygenation-induced neuronal injury in mouse hippocampus

作　　者：魏健强[1] 张春瑞[2] WEI Jianqiang;ZHANG Chunrui(Neurovascular Interventional Therapy Center,the Affiliated Hospital of Yan'an University,Yan'an 716000;Department of Neurology,Hanzhong People's Hospital,Hanzhong 723000,China)

机构地区：[1]延安大学附属医院神经血管介入治疗中心,陕西延安716000 [2]汉中市人民医院神经内科,陕西汉中723000

出　　处：《基础医学与临床》2023年第2期252-258,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(81701207)。

摘　　要：目的探讨咪达唑仑基于丝裂原活化蛋白激酶(MAPK)/核转录因子-κB(NF-κB)信号通路对缺氧/复氧诱导的神经元损伤的影响。方法体外培养小鼠海马神经元,MTT法检测0、5、10、40、70和100 ng/mL咪达唑仑对其缺氧/复氧后细胞活力的影响,筛选出最佳作用浓度。将培养的小鼠海马神经元随机分为对照组、模型组(缺氧3 h再复氧12 h)、咪达唑仑(70 ng/mL)组、C16-PAF(MAPK激活剂,4μmol/L)组、咪达唑仑(70 ng/mL)+C16-PAF组(4μmol/L),通过Hoechst33258染色和流式细胞检测神经元凋亡情况;ELISA测量细胞乳酸脱氢酶(LDH)、肿瘤坏死因子-α(TNF-α)、白细胞介素-18(IL-18)、白细胞介素-17(IL-17)释放量及细胞超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性;免疫印迹检测细胞凋亡蛋白及MAPK/NF-κB通路蛋白表达。结果与对照组比较,模型组神经元凋亡率、LDH释放量、MAPK/NF-κB通路p-p38 MAPK/p38 MAPK、p-NF-κB p65/NF-κB p65水平明显升高(P<0.05)。与模型组、咪达唑仑+C16-PAF组分别比较,咪达唑仑组神经元凋亡率、LDH释放量、MAPK/NF-κB通路p-p38 MAPK/p38 MAPK、p-NF-κB p65/NF-κB p65水平均降低(P<0.05);C16-PAF组神经元凋亡率、LDH释放量、MAPK/NF-κB通路p-p38 MAPK/p38 MAPK、p-NF-κB p65/NF-κB p65水平均升高(P<0.05)。结论咪达唑仑可能通过抑制MAPK/NF-κB信号激活降低炎性细胞因子表达,进而抑制缺氧/复氧诱导的炎性和氧化应激反应,减轻神经元损伤,降低其凋亡率。Objective To investigate the impact of midazolam on hypoxia-reoxygenation-induced neuronal injury based on mitogen-activated protein kinase(MAPK)/nuclear factor-κB(NF-κB)signaling pathway.Methods Mouse hippocampus neurons were cultured in vitro and the effects of 0,5,10,40,70 and 100 ng/mL midazolam on cell viability after hypoxia and reoxygenation were detected by MTT assay,and the best concentration was screened out.The cultured mouse hippocampus neurons were randomly separated into control group,model group(anoxic for 3 hours and reoxygenated for 12 hours),midazolam(70 ng/mL)group,C16-PAF(MAPK activator,4μmol/L)group,and midazolam(70 ng/mL)+C16-PAF group(4μmol/L).The apoptosis of neurons was detected by Hoechst33258 staining and flow cytometry;the releases of lactate dehydrogenase(LDH),tumor necrosis factor-α(TNF-α),interleukin-18(IL-18),and interleukin-17(IL-17)and the levels of cellular superoxide dismutase(SOD)and catalase(CAT)were detected by ELISA;the expression of apoptosis proteins and MAPK/NF-κB pathway proteins were detected by western blotting.Results Compared with the control group,the neuron apoptosis rate,LDH,the TNF-α,IL-18,IL-17 releases,apoptosis protein caspase-9,Bax expression,and MAPK/NF-κB pathway p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 levels were significantly increased in the model group(P<0.05),the CAT and SOD levels in cells were significantly decreased(P<0.05).Compared with the model group and midazolam+C16-PAF group,the neuron apoptosis rate,LDH,the TNF-α,IL-18,IL-17 releases,apoptosis protein caspase-9,Bax expression,and MAPK/NF-κB pathway p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 levels were significantly decreased in the midazolam group(P<0.05),the CAT and SOD activity in cells were significantly increased(P<0.05);the neuron apoptosis rate,LDH,the TNF-α,IL-18,IL-17 releases,apoptosis protein caspase-9,Bax expression,and MAPK/NF-κB pathway p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 levels were significantly increased in the C16-PAF group(P<0.05),the

关 键 词：咪达唑仑 丝裂原活化蛋白激酶/核转录因子-κB 缺氧/复氧 神经元 损伤 

分 类 号：R285.5[医药卫生—中药学] R743.3[医药卫生—中医学]