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

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50 ml,加入20 U/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%CO_2培养箱中培养12 h;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5 μg/ml)、异丙酚(终浓度为5 μg/ml)或LPS(终浓度为1μg/ml),置于37℃、5%CO_2培养箱中孵育12 h;intralipid+LPS组(IL组)和异丙酚+LPS组(PL组)先分别加入intralipid(终浓度为5 μg/ml)或异丙酚(终浓度为5 μg/ml)后,置于37℃、5%CO_2培养箱中孵育20 min,然后加入LPS(终浓度为1μg/ml),置于37℃、5%CO_2培养箱孵育12 h.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PGR检测中性粒细胞膜TLR2 mRNA和TLR4 mRNA的表达;采用ELISA法测定培养上清液TNF-α和IL-8的浓度.结果 与C组比较,I组和P组TLB2和TLR4的表达、1NF-α和IL-8L-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IL-8L-8的浓度升高,IL组IL-8浓度升高(P<0.05);与L组比较,IL组TLR2和TLR4的表达、TNF-α和IL-8L-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-8L-8的浓度降低(P<0.05).各组TLR2 mRNA和TLR4 mRNA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.

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

Objective To investigate the effect of propofol on the Toll like receptor 2 (TLR2) and TLR4 expression on lipopolysaccbaride (LPS)-activated polymorpbanuclear 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 intraipid (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 1 μg/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 concentratoons in the supernatant were analyzed by ELISA.Results LPS significantly up-regulated TLR2 and TLR4 expression on the membrane of PMNs and increased TNF-αand 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-indnced up-regulation of TLR2 and TLR4 expression on PMNs, resulting in suppression of systemic inflammatory response mediated by PMNs.