摘要
目的 探讨右美托咪定（dexmedetomidine,Dex）对脂多糖（lipopolysaccharides,LPS）诱导单核巨噬细胞RAW264.7炎性因子释放及血红素氧合酶（heme oxygenase,HO）-1表达的影响. 方法 离体培养小鼠单核巨噬细胞系RAW264.7,用含有1×106个/ml的细胞样本接种于6孔培养板（2 ml/孔）.采用随机数字表法将其分为5组（每组6孔）：正常对照组（C组）、Dex 10 μg/L组（Dex组）、LPS 1 mg/L组（LPS组）、LPS 1 mg/L＋Dex 10 μg/L组（LPS＋Dex组）及HO-1抑制剂ZnPPⅨ10 μmol/L＋LPS 1 mg/L＋Dex 10 μg/L组（ZnPPⅨ＋LPS＋Dex组）.分别于孵育3、6、12、24 h时点收集细胞培养上清液,采用ELISA法测定TNF-α、IL-6、高迁移率族蛋白1（high mobility group box-1 protein,HMGB-1）的浓度;在6 h和12 h时点收集细胞沉淀,提取蛋白成分,利用Western blot法测定HO-1表达. 结果 与C组比较,LPS组各时间点TNF-α、IL-6、HMGB-1浓度均明显升高（P〈0.05）,在6、12 h时点细胞HO-1的表达上调（P〈0.05）;与LPS组比较,LPS＋Dex组各时间点的TNF-α、IL-6、HMGB-1浓度降低（P〈0.05）,培养细胞6、12 h时点HO-1表达升高;与LPS＋Dex组比较,ZnPPⅨ＋ LPS＋Dex组各时间点的TNF-α、IL-6、HMGB-1浓度升高（P〈0.05）,培养细胞6、12 h时点HO-1表达降低（P〈0.05）. 结论 Dex可以抑制LPS诱导的单核巨噬细胞炎症因子释放,其机制与上调HO-1表达有关.
Objective To explore the effect of dexmedetomidine （Dex） on the release of cell cytokines from monocyte macrophage RAW264.7 cells induced by lipopolysaccharide （LPS） and the expression of heme oxygenase （HO）-1. Methods RAW264.7 cells were cultured in six-well culture dishes with a density of 1×106/ml（2 ml/hole）and were randomly divided into 5 groups（n=6）by the number table method：normal control group（C group）,Dex 10 μg/L（Dex group）,LPS 1 mg/L（LPS group）,LPS 1 mg/L＋Dex 10 μg/L （LPS＋Dex group）,and ZnPPⅨ （inhibitor of HO-1）,10 μmol/L＋LPS 1 mg/L＋Dex,10 μg/L （ZnPPⅨ＋LPS＋Dex group）. After incubation of 3, 6, 12, 24 h, ELISA was used to determine the concentrations of TNF-α, IL-6 and high mobility group box-1 protein（HMGB-1）in the supernatants of six wells in each group.Then,six wells in each group were chosen at incubation of 6 h and 12 h for determination of HO-1 expression by Western blot. Results Comparing with C group, the concentrations of TNF-α, IL-6 and HMGB-1 in the supernatants at 3,6,12,24 h in the LPS group were significantly increased （P〈0.05）,and the expression of HO-1 at 6 h and 12 h time point was up-regulated in LPS group. Compared with LPS group, the concentrations of TNF-α, IL-6 and HMGB-1 in LPS＋Dex group were attenuated（P〈0.05）.Meanwhile,the expression of HO-1 at 6 h and 12 h time point was up-regulated in LPS＋Dex group.Compared with LPS＋Dex group,the concentrations of TNF-α,IL-6 and HMGB-1 in ZnPPⅨ＋LPS＋Dex group were increased （P〈0.05）. However,the expression of HO-1 was reduced in ZnPPⅨ＋LPS＋Dex group （P〈0.05）. Conclusions Dex could inhibit the release of cell cytokines from RAW264.7 via up-regulating the levels of HO-1 expression.
作者
陆季娟
杭黎华
Lu Jijuan, Hang Lihua(Department of Anesthesiology, Kunshan First People＇s Hospital Affiliated to Jiangsu University, Kunshan 215300, China （Lu JJ, Hang LH）)
出处
《国际麻醉学与复苏杂志》
CAS
2018年第2期136-138,169,共4页
International Journal of Anesthesiology and Resuscitation
关键词
右美托咪定
炎症因子
脂多糖
血红素氧合酶-1
Dexmedetomidine
Inflammatory cytokines
Lipopolysaccharide
Heme oxygenase-1
