TLR4／NF-κB信号通路与丙泊酚抑制内毒素诱导大鼠肺泡巨噬细胞TNF-α释放的关系  

Relationship between TLR4/NF-κB signaling pathway and propofol-induced inhibition of endotoxin- induced release of TNF-α from alveolar macrophages of rats

作　　者：杨雪[1] 孙赳 曾思[2] 兰志勋[2] 

机构地区：[1]宜宾市第一人民医院麻醉科,644000 [2]四川省人民医院麻醉科,成都市610000

出　　处：《中华麻醉学杂志》2017年第6期761-764,共4页Chinese Journal of Anesthesiology

摘　　要：目的评价Toll样受体4（TLR4）／NF—κB信号通路与丙泊酚抑制内毒素诱导大鼠肺泡巨噬细胞TNF-α释放的关系。方法成年雄性SD大鼠，提取、培养肺泡巨噬细胞，接种于6孔板（1×106个/孔）和96孔板（1×104个／孔）。采用随机数字表法分为5组（n=18）：对照组（C组）PBS继续培养；二甲基亚砜组（D组）加入二甲基亚砜，终浓度5mg／ml；LPS组（L组）加入LPS，终浓度1Ixg／ml；丙泊酚组（P组）加入丙泊酚，终浓度25μmol／L（4．46μg／m1）；LPS＋丙泊酚组（L＋P组）加入LPS和丙泊酚，终浓度分别为1μg／ml和25μmol／L（4．46p,g／m1）。培养或孵育24h时，采用CCK-8法检测细胞活力，采用瑞氏染色观察细胞形态变化，采用ELISA法检测上清液TNF—α浓度，采用Westernblot法检测TLR4表达和NF—κB活性。结果与c组比较，L组、L＋P组细胞活力和上清液TNF—α浓度升高，TLR4表达上调，NF—κB活性增强（P〈0．05），D组和P组上述指标差异无统计学意义（P〉0．05）；与L组比较，L＋P组细胞活力和上清液TNF—α浓度降低，TLR4表达下调，NF—κB活性减弱（P〈0．05），细胞形态变化减轻，伸出的伪足数减少。结论丙？白酚抑制内毒素诱导大鼠肺泡巨噬细胞TNF—α释放的机制与抑制TLR4／NF—κB信号通路激活有关。Objective To evaluate the relationship between Toll-like receptor 4 （TLR4）/nuclear factor kappa B （NF-KB） signaling pathway and propofol-induced inhibition of endotoxin-induced release of tumor necrosis factor-alpha （TNF-α） from alveolar maerophages （AMs） of rats. Methods AMs extrac- ted from adult male Sprague-Dawley rats were cultured and inoculated in 6-well plates （ 1× 106 cells/well） and in 96-well plates （ 1×104 cells/well）. The cells were divided into 5 groups （n= 18 each） using a ran- dom number table： control group （ group C） , dimethyl sulfoxide group （ group D） , lipopolysaccharide （LPS） group （group L）, propofol group （group P） and LPS plus propofol group （group L＋P）. The cells were continuously cultured with phosphate buffer solution in group C. Dimethyl sulfoxidc was added at the final concentration of 5 mg/ml in group D. LPS was added at the final concentration of 1 μg/ml in group L. Propofol was added at the final concentration of 25 μmol/L （4.46 μg/ml） in group P. LPS and propo- fol were added at the final concentration of 1 μg/ml and 25 μmol/L （4.46 μg/ml） , respectively, in group L＋P. At 24 h of culture or incubation, the cell viability was detected by CCK-8 assay, the morphological changes of cells were observed using Wright＇s staining, the concentration of TNF-α in the supernatant was determined by enzyme-linked immunosorbent assay, and TLR4 expression and NF-κB activities were meas-ured by Western blot. Results Compared with group C, the cell viahilily and concentration of TNF-α in the supernatant were significantly increased, the expression of TLR4 was up-regulated, and the activity of NF-κB was enhanced in L and L＋P groups （P〈0. 05） , and no significant change was tbund in the parame- ters mentioned above in 1） and P groups （P〉0. 05）. Compared with group L, the cell viability and concen- tration of TNF-α in the supernatant were significantly decreased, the expression of TLR4 was down-regula- t

关 键 词：Toll样受体4 NF-ΚB 二异丙酚 内毒素类 巨噬细胞 肺泡 肿瘤坏死因子α 

分 类 号：R614[医药卫生—麻醉学]