异丙酚对LPS诱导中性粒细胞Toll样受体2和Toll样受体4表达的影响  

Effect of propofoi on Toll like receptor-2 and 4 expression on LPS-activated polymorphonuclear leukocytes

作　　者：程楠[1] 池信锦[1] 黑子清[1] 罗刚健[1] 蔡君[1] 

机构地区：[1]中山大学附属第三医院麻醉科,广州市510630

出　　处：《中华麻醉学杂志》2009年第10期911-914,共4页Chinese Journal of Anesthesiology

基　　金：广东省自然科学基金资助项目（7001567）;广东省科技计划项目（2007B031511009）

摘　　要：目的 探讨异丙酚对内毒素（LPS）诱导中性粒细胞Toll样受体2（TLR2）和TLR4表达的影响。方法健康志愿者6名，年龄20～35岁，各采集外周静脉血样50ml，加入20U／ml肝素抗凝，分离提纯中性粒细胞，制备细胞悬液，然后随机分为6组，每组6皿：对照组（C组）不给予任何药物，置于37℃、5％CO2培养箱中培养12h；异丙酚脂质溶剂intralipid组（I组）、异丙酚组（P组）和LPS组（L组）：分别加入intralipid（终浓度为5μg／ml）、异丙酚（终浓度为5Itg／m1）或LPS（终浓度为1ttg／m1），置于37℃、5％CO，培养箱中孵育12h；intralipid＋LPS组（IL组）和异丙酚＋LPS组（PL组）先分别加入intralipid（终浓度为5μg／ml）或异丙酚（终浓度为5μg／ml）后，置于37℃、5％CO2培养箱中孵育20min，然后加入LPS（终浓度为1μg／ml），置于37℃、5％CO2培养箱孵育12h。采用流式细胞仪测定中性粒细胞膜TLR2和TLR4的表达；采用荧光定量PCR检测中性粒细胞膜TLR2mRNA和TLR4mRNA的表达；采用ELISA法测定培养上清液TNF-α和11-8的浓度。结果与C组比较，I组和P组TLR2和TLR4的表达、TNF-α和11-8的浓度差异无统计学意义（P〉0．05），L组和IL组TLR2和TLR4的表达上调，L组，TNF-α和11-8的浓度升高，IL组11-8浓度升高（P〈0．05）；与L组比较，IL组TLR2和TLR4的表达、TNF—α和21-8的浓度差异无统计学意义（P〉0．05），PL组TLR2和TLR4的表达下调，TNF-α和IL-8的浓度降低（P〈0．05）。各组TLR2mRNA和TLR4mRNA的表达差异无统计学意义（P〉0．05）。结论异丙酚可下调LPS诱导的中性粒细胞TLR2和TLR4的表达，从而抑制炎性反应。Objective To investigate the effect of propofol on the Toll like receptor 2 （TLR2） and TLR4 expression on lipopolysaccharide （LPS）-activated polymorphonuclear leukocytes （PMNs）. Methods PMNs were isolated from the venous blood collected from 6 healthy volunteers aged 20-35 yr and incubated in RPMI 1640 liquid cell culture medium. The cultured PMNs were randomly divided into 6 groups （ n = 6 each） ： group control （group C）; group intralipid （group I）; group propofol （group P）： the PMNs were incubated in the presence of propofol 5 μg/ml; group LPS （group L）： the PMNs were exposed to LPS 1μg/ml; LPS （1 μg/ml） ＋ intralipid （group IL） and LPS （1 μg/ml） ＋ propofol （5 μg/ml）（ group PL）. After 12 h incubation, the TLR2 and TLR4 expression on the membrane of PMNs was measured by flow cytometry. The TLR2 mRNA and TLR4 mRNA expression was assessed by RT-PCR. TNF-α and IL-8 concentrations in the supernatant were analyzed by ELISA. Results LPS significantly up-regulated TLR2 and TLR4 expression on the membrane of PMNs and increased TNF-α and IL-8 concentrations in the supernatant. Propofol significantly inhibited the LPS-induced changes. There was no significant difference in TLR2 and TLR4 mRNA expression among the 6 groups. Conclusion Propofol can inhibit LPS-induced up-regulation of TLR2 and TLR4 expression on PMNs, resulting in suppression of systemic inflammatory response mediated by PMNs.

关 键 词：二异丙酚 TOLL样受体2 TOLL样受体4 内毒素类 中性白细胞 

分 类 号：R96[医药卫生—药理学]