右美托咪啶减轻LPS诱导的巨噬细胞炎症反应及其对NF-κB信号通路的影响  

Role of dexmedetomidine in lipopolysaccharide-induced macrophage inflammatory response and nuclear transcription factor-κB signal pathway

作　　者：张念亮[1] 刘祥兰[2] 孙波[3] 单海华[3] 

机构地区：[1]济宁医学院附属日照市人民医院麻醉科,日照276800 [2]济宁医学院附属日照市人民医院心脏工作站,日照276800 [3]苏州大学附属第二医院麻醉科,苏州215004

出　　处：《中华神经医学杂志》2017年第6期581-584,共4页Chinese Journal of Neuromedicine

基　　金：江苏省自然科学基金（BK20141187）;苏州市科技计划项目fSYS201039）

摘　　要：目的探讨右美托咪啶（DFX）对脂多糖（LPS）诱导的单核巨噬细胞RAW264．7炎性因子释放和核转录因子-κB（NF-κB）信号通路的影响。方法将体外常规培养的小鼠巨噬细胞株RAW264．7分为正常对照组、DEX组、LPS组、LPS＋DEX组，DEX、LPS的终浓度分别为10ng／mL、1μg／mL，正常对照组加入等量PBS。继续培养3、6、12、24h时，ELISA检测各组细胞炎性因子肿瘤坏死因子-α（TNF-α）、白介素（IL）-1β、高迁移率族蛋白1（HMGB-）的释放；培养12h时，Westernblotting检测各组细胞NF-κB信号通路相关蛋白磷酸化（p）-P65、P-P50的表达。结果（1）细胞培养3、6、12、24h时，与正常对照组比较，LPS组细胞培养上清液TNF-α、IL-6、HMGB-1浓度升高，差异有统计学意义（P〈0．05）；与LPS组比较，DEX＋LPS组细胞培养上清液TNF-α、IL-1β、HMGB-1浓度降低，差异均有统计学意义（P〈0．05）。（2）Westernblotting检测显示细胞培养12h时，与正常对照组比较，LPS组细胞P-P65、P-P50蛋白表达量升高，差异均有统计学意义（P〈0．05）。与LPS组比较，DEX＋LPS组细胞P-P65、P-P50蛋白表达量降低，差异有统计学意义（P〈0．05）。结论DEX可以抑制LPS诱导的单核巨噬细胞炎症因子的释放，降低NF-κB信号通路相关蛋白P-P65和P-P50的表达。Objective To investigate the role of dexmedetomidine （DEX） in lipopolysaccharide （LPS）-induced release of cell cytokines from monocyte macrophage RAW264.7 and nuclear transcription factor （NF）-κB signal pathway. Methods RAW264.7 cell lines were seeded in 6-well plates with a density of1×10^6/mL （2 mL/hole） and randomly divided into control group （PBS）, DEX group （10 ng/mL）, LPS group （1 μg/mL） and LPS （1 μg/mL）＋DEX （10 ng/mL） group. After incubation of 3, 6, 12 and 24 h, ELISA was employed to detect the concentrations of tumor necrosis factor （TNF）-α, interleukin （IL）-1 and high mobility group box （HMGB）-1 in the supernatants of 6 wells in each group. Then, 6 wells in each group were chosen at incubation of 12 h for determination of NF-κB signal pathway related proteins phosphorylated （p）-P65 and p-P50 expressions by Western blotting. Results As compared with those in the control group, the concentrations ofTNF-cq IL-1β and HMGB-1 in the supematant of LPS group were significantly increased （P〈0.05）, and the expressions of p-P65 and p-P50 were significantlyincreased in the LPS group （P〈0.05）. As compared with those in the LPS group, the levels of TNF-α, IL-1β and HMGB-1 were statistically decreased, and the expressions of P65 and P50 were significantly down-regulated in the DEX group （P〈0.05）. Conclusion DEX could inhibit the release of cells cytokines from RAW264.7 and decrease the levels of NF-κB signal pathway related protein expressions.

关 键 词：右美托咪啶 脓毒症 炎症因子 核转录因子-κB信号通路 

分 类 号：R614[医药卫生—麻醉学]