异丙酚预先给药对BV-2细胞缺氧复氧时TLR4表达的影响

Effects of propofol pretreatment on Toll-like receptor 4 in BV-2 cells after anoxia-reoxygenation

目的评价异丙酚对BV-2细胞缺氧复氧时Toll样受体4（TLR4）表达的影响。方法小鼠小胶质细胞（BV-2细胞）于6孔培养板中培养4～6d后随机分为4组（n=4），正常对照组（C组）不给予任何处理；缺氧复氧组（A／R组）缺氧3h、复氧12h；缺氧复氧＋25μmol／L异丙酚组（P25组）和缺氧复氧＋100μmol／L异丙酚组（P100组）于缺氧前30min加入异丙酚，终浓度分别为25、100μmol／L。于复氧12h时收集细胞，测定TLR4 mRNA、NF-κB mRNA和TLR4蛋白水平；收集细胞上清液，测定TNF-α水平。结果与C组比较，A／R组、P25组和P100组TLR4 mRNA、NF-κB mRNA、TLR4蛋白和TNF-α水平均升高（P〈0．01）；与A／R组比较，P25组和P100组TLR4 mRNA、NF-κB mRNA、TLR4蛋白和TNF-α水平均下降（P〈0．01）；与P25组比较，P100组TLR4 mRNA、NF-κB mRNA、TLR4蛋白和TNF-α水平均降低（P〈0．01）。结论异丙酚25、100μmol／L预先给药可抑制BV-2细胞缺氧复氧时TLR4 mRNA表达上调。

Objective To investigate the effects of propofol pretreatment on the expression of Toll-like receptor 4 in BV-2 cells following anoxia-reoxygenation （A/R）. Methods BV-2 mouse microglial cells purchased from PUMC cell bank were cultured in 6-well plates and randomly divided into 4 groups （ n = 4 each） ： group Ⅰ control （C） ; group Ⅱ A/R; group Ⅲ and Ⅳ 25 and 100 μmol/L propofol pretreatment （P25, P100 ） . In group A/R （ Ⅱ ） The cells were exposed to 95% N2-5% CO2 in an incubator at 37℃ for 3 h followed by reoxygenation with 95% 02-5% CO2 for 12 h. In group P25 and P100 propofol was added to the culture medium with the final concentration of 25 and 100 μmol/L respectively at 30 min before anoxia. At the end of 12 h reoxygenation the expression of TLR4 protein, TLR4 mRNA and NF-κB mRNA of BV-2 ceils and the TNF-α level in the supernatant were measured. Results The expression of TLR4 protein and mRNA and NF-κB mRNA of BV-2 ceils and the TNF-α level in the supernatant were significantly higher in group A/R, P25 and P100 than in group C （P 〈 0.01 ）. The expression of TLR4 protein and TLR4 mRNA and NF-κB mRNA were significantly decreased by propofol pretreatmcnt in a dose-dependent manner in group P25 and P100 as compared to A/R group. Conclusion Propofol 25,100 μmol/L pretreatment can inhibit the up-regulation of TLR4 mRNA expression in BV-2 cells induced by A/R.