丙泊酚对小胶质细胞炎症因子的影响及其机制研究

Effect of Propofol on inflammatory cytokines in microglia and its mechanism

摘要:

目的 探讨丙泊酚对小胶质细胞炎症因子的影响及其机制。方法 将小胶质BV-2 细胞分为对照组、丙泊酚组、脂多糖（LPS）组、LPS+ 丙泊酚组，对照组细胞加入PBS 液，丙泊酚组细胞加入丙泊酚30μmol/L，LPS 组细胞加入LPS 1μg/ml，LPS+ 丙泊酚组细胞加入丙泊酚30μmol/L+LPS 1μg/ml。采用MTT 比色实验测定细胞活性，采用酶联免疫吸附法测定细胞上清液中白细胞介素1β（IL-1β）、白细胞介素6（IL-6）、肿瘤坏死因子-α（TNF-α）水平，采用逆转录- 聚合酶链反应测定细胞p38MAPK 和TLR4mRNA 的表达，采用Western blot 检测细胞p38MAPK 和TLR4 蛋白表达量。结果 LPS 组和LPS+ 丙泊酚组小胶质细胞活性低于对照组和丙泊酚组（P <0.05），LPS+ 丙泊酚组小胶质细胞活性高于LPS 组（P <0.05）；LPS 组和LPS+ 丙泊酚组小胶质细胞上清液中IL-1β、IL-6、TNF-α 水平高于对照组和丙泊酚组（P <0.05），LPS+ 丙泊酚组小胶质细胞上清液中IL-1β、IL-6、TNF-α 水平低于LPS 组（P <0.05）；LPS 组和LPS+ 丙泊酚组小胶质细胞p38MAPK、TLR4 mRNA 和蛋白表达量高于对照组和丙泊酚组（P <0.05），LPS+ 丙泊酚 组小胶质细胞p38MAPK、TLR4 mRNA 和蛋白表达量低于LPS 组（P <0.05）；丙泊酚组和对照组小胶质细胞各指标比较，差异无统计学意义（P >0.05）。结论 丙泊酚能抑制小胶质细胞过度活化和炎症反应，其机制可能与丙泊酚可下调TLR4-p38MAPK 信号通路有关。

Abstract:

Objective To investigate the effect of Propofol on inflammatory cytokines in microglia and its mechanism. Methods Microglial BV-2 cells were divided into control group, Propofol group, lipopolysaccharide (LPS) group, and LPS+Propofol group. The cells of the control group were added with PBS. The cells of the Propofol group were added with 30 μmol/L Propofol. The cells of the LPS group were treated with 1 μg/ml LPS. The cells of the LPS+Propofol group were added with 30 μmol/L Propofol and 1 μg/ml LPS. The cell viability was determined by MTT colorimetric assay. The levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in supernatant were measured by ELISA. The expressions of p38MAPK and TLR4 mRNAs were detected by RT-PCR.The expressions of p38MAPK and TLR4 proteins were detected by Western blot. Results The activity of microglia in the LPS group and the LPS+Propofol group were lower than that in the control group and the Propofol group(P < 0.05). The activity of microglia in the LPS+Propofol group was higher than that in the LPS group (P < 0.05). The levels of IL-1β, IL-6 and TNF-α in the supernatant of the LPS group and the LPS+Propofol group were higher than those in the control group and the Propofol group (P < 0.05). The levels of IL-1β, IL-6 and TNF-α in the supernatant of the LPS+Propofol group were lower than those in the LPS group (P < 0.05). The mRNA and protein expressions of p38MAPK and TLR4 in the LPS group and the LPS+Propofol group were higher than those in the control group and the Propofol group (P < 0.05). The mRNA and protein expressions of p38MAPK and TLR4 in the LPS+Propofol group were lower than those in the LPS group (P < 0.05). There were no significant differences in the above indices between the Propofol group and the control group (P > 0.05). Conclusions Propofol has the effect of inhibiting the hyperactivity and inflammatory response of microglia, and its mechanism may be related to the downregulation of TLR4-p38MAPK signaling pathway.