丙泊酚对脂多糖诱导的BV2细胞中炎性因子表达的影响  

Effect of Propofol on the Expression of Inflammatory Factors in LPSinduced BV2 Cells

作　　者：徐雅洁[1] 刘争杰 董诺亚 XU Yajie;LIU Zhengjie;DONG Nuoya(Department of Anesthesiology,Port Hospital of Hebei Port Group Co.,Ltd.,Qinhuangdao,Hebei,066002,China)

机构地区：[1]河北港口集团有限公司港口医院麻醉科,河北省秦皇岛市066002

出　　处：《医学分子生物学杂志》2023年第4期351-358,共8页Journal of Medical Molecular Biology

基　　金：2021年秦皇岛市科学技术研究与发展计划项目(No.202101A088)。

摘　　要：目的探讨丙泊酚(propofol)通过调控核转录因子-κB(nuclear transcription factor-κB,NF-κB)信号通路对活化的BV2细胞炎性因子的影响。方法将BV2细胞分为5组:对照组、模型组、丙泊酚低剂量组(20μmol/L)、丙泊酚中剂量组(50μmol/L)和丙泊酚高剂量组(100μmol/L)。CCK-8(cell counting Kit-8)实验检测细胞活力;ELISA(enzyme linked immunosorbent assay)实验检测白细胞介素-6(interleukin-6,IL-6)、IL-1β、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和一氧化氮合酶(nitric oxide synthase,NOS)水平;试剂盒检测活性氧(reactive oxygen species,ROS)活性;Western印迹检测核苷酸结合寡聚化结构域样受体蛋白3(NOD-like receptor protein 3,NLRP3)、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)、半胱氨酸蛋白酶-1(Caspase-1)、IκB和P65的表达及P65的磷酸化水平;免疫荧光染色观察P65进入胞核的情况。结果3种浓度丙泊酚对细胞活力无影响。与对照组比较,LPS组的IL-6、IL-1β、TNF-α、ROS和NOS水平显著上升(P<0.01);IκB的表达显著下调,p-P65/P65的表达显著上调,P65入核表达显著增多(P<0.01);经过不同剂量丙泊酚处理BV2细胞后,逆转了上述结果,IL-6、IL-1β、TNF-α、ROS和NOS水平显著降低(P<0.01);IκB的表达显著上调,p-P65/P65的表达显著下调,核定位的P65蛋白显著减少(P<0.01),且上述结果呈剂量相关性。结论丙泊酚可通过调节NF-κB信号通路减缓活化的BV2细胞中的炎性因子的表达。Objective To investigate the effect of propofol on inflammatory cytokines activation by regulating the nuclear factor kappa-B(NF-κB)signaling pathway in BV2 microglial cells.Methods BV2 cells were divided into 5 groups:control group,model group,propofol lowdose group(20μmol/L),propofol medium-dose group(50μmol/L),and propofol high-dose group(100μmol/L).Cell viability was detected by CCK-8 assay.The levels of interleukin-6(IL-6),IL-1β,tumor necrosis factor-α(TNF-α),nitric oxide synthase(NOS)were detected by ELISA,and the reactive oxygen species(ROS)was detected by ROS assay kit.Western blotting analysis was used to detect the expression levels of NOD-like receptor protein 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC),Caspase-1,IκB,P65,and p-P65.Immunofluorescence staining was used to observe the nuclear translocation of P65.Results Propofol showed no toxicity to BV3 cells for all three concentrations.The levels of IL-6,IL-1β,TNF-α,ROS,and NOS in the LPS group were significantly increased when compared with those in the control group(P<0.01).The expression of IκB was significantly down-regulated,p-P65/P65 was significantly up-regulated,and P65 nuclear translocation was significantly increased(P<0.01).After treatment with different doses of propofol,the results of above indexes were reversed,IL-6,IL-1β,TNF-α,ROS and NOS levels were significantly reduced(P<0.01),the expression of IκB was significantly up-regulated,p-P65/P65 was significantly down-regulated,and nuclear localized p65 protein was significantly reduced(P<0.01).All changes in the above results were dose-dependent.Conclusion Propofol alleviate the activation of inflammatory cytokines in LPS stimulated BV2 cells by regulating the NF-κB signaling pathway.

关 键 词：丙泊酚 NF-ΚB 炎性因子 BV2细胞 

分 类 号：R741.05[医药卫生—神经病学与精神病学]