异丙酚对缺氧复氧大鼠海马星形胶质细胞的影响

Effects of Propofol on Primary Cultured Hippocampal Astrocytes with Anoxia and Re-oxygenation in Rats

目的:观察异丙酚对缺氧复氧大鼠海马星形胶质细胞及其分泌促炎性细胞因子(TNF-α、IL-1β、IL-6)和抗炎性细胞因子IL-10变化的影响。方法:取新生2～3dWistar大鼠海马星形胶质细胞,原代纯化培养3周。将细胞分为正常对照组(Nor组),乳剂对照组(Nor-L组),缺氧6h后复氧24h组(Ano组),加异丙酚50μmol/L缺氧6h后复氧24h组(Pro1组)、加异丙酚500μmol/L缺氧6h后复氧24h组(Pro2组)。取各组培养孔上清液检测IL-1β、TNF-α、IL-6和IL-10含量,用神经胶质纤维酸性蛋白(GFAP)免疫细胞化学法标记星形胶质细胞,用多媒体彩色病理图像分析系统检测阳性细胞平均光密度(AOD)。结果:与Nor组比较,Ano组细胞被激活增生肥大,AOD升高(P<0.01),其上清液中IL-1β、IL-6和TNF-α含量上升(P<0.01),IL-10无明显变化(P>0.05)。与Ano组比较,Pro1组和Pro2组细胞激活被抑制,AOD、IL-1β、IL-6和TNF-α含量均降低(其中Pro1组P<0.05,Pro2组P<0.01),而IL-10含量均上调(P<0.01)。Nor组和Nor-L组比较无明显差异。结论:异丙酚可抑制缺氧复氧大鼠海马星形胶质细胞的激活作用,同时抑制星形胶质细胞分泌IL-1β、IL-6和TNF-α,增强IL-10的合成释放,其作用程度与剂量有关。

Objective To investigate the effects of propofol on primary cultured hippocampal astrocytes with anoxia and reoxygenation in rats and the changes of interleukin-1 peptide（ IL-1 β） ,tumor necrosis Factor-α（ TNF-α）, IL-6 and IL-10 in response to anoxia and re-oxygenation. Methods Hippocampal astrocytes isolated and purified from neonatal wistar rats （ 2 3 days） were cultured for 3 weeks. The cells were treated as follows： normal control group：sham wash with Hanks solution（ group Nor） ; Lipid vehicle group： intralipid 0.2 ml/L added（ group Nor-L） ;6 h anoxia and 24 h re-oxygenation group （Group Ano） ;6 h anoxia and 24 h re-oxygenation after added propofol 50μmol/L group （Group Pro1 ） ;6h anoxia and 24h re-oxygenation after added propofol 500μmol/L group （Group Pro2 ）. Then determined the concentrations of IL-1μ,TNF-α, IL-6 and IL-10 in the supernatant,labelled the cells using GFAP immunostaining and analyzed the average optical density （AOD） with multimedia pathology imaging analysis system（ MPIAS）. Results Compared with group Nor,the cells were activated and hypertrophic, AOD were heightened and the concentrations of IL-1β, TNF-α and IL-6 were also significantly increased in Group Ano （P 〈 0.01 ）, but there was no significant change in IL- 10 （P 〉 0.05 ）. Compared with Group Ano, the activation of cells was inhibited by propofol, the AOD, IL-1β, TNF-α and IL-6 were significantly lower in the Pro1 group （ P 〈 0.05 ） and Pro2 group（ P 〈 0.01 ）, while IL-10 was greatly higher in the two groups （ P 〈 0.01 ）. There was no significant difference between control group and intralipid vehicle group. Conclusions Propofol can effectively inhibit the activation of astrocytes in hippocampus induced by anoxic and re-oxygenation,it also can significantly inhibit the production of IL- 1β, TNF-α and IL-6, enhance synthesis and release of IL-10, its extent of action is related to the dose administered.