促血小板生成素在小胶质细胞炎症模型中的表达与信号通路研究

Signal transduction pathway mediated by thrombopoietin in the inflammation model of microglia

摘要:

目的：初步探讨促血小板生成素（thrombopoietin, TPO）在脂多糖（lipopolysaccharide, LPS）诱导小胶质细胞炎症模型中的信号转导通路。方法：以0.5和1.0 μg/mL浓度的LPS刺激体外培养小鼠小胶质细胞（BV2细胞）建立炎症模型，采用实时荧光定量PCR法检测细胞内TPO及ERK mRNA水平；Western blot法检测TPO及ERK 蛋白水平；使用ELISA方法测定细胞培养液中IL-6和TPO的含量。结果：LPS干预后能显著增加BV2细胞内TPO及ERK mRNA和蛋白水平的表达，尤其以浓度为1.0 μg/mL的LPS干预12 h 时，二者的表达水平达到最高峰；且二者的表达呈显著正相关。结论：ERK1/2信号转导通路参与了BV2细胞炎症损伤中的TPO的活化作用。

Abstract:

OBJECTIVE: To explore the signal transduction pathway mediated by thrombopoietin (TPO) in the inflammation model of microglia induced by lipopolysaccharide (LPS). METHODS: The inflammation model of microglia BV2 cells was prepared by LPS of 0.5 and 1.0 μg/mL stimulation. The expression of TPO and ERK mRNA in BV2 cells was detected by real time quantitative PCR. Western blot was used to evaluate the expression of TPO and ERK protein in BV2 cells. TPO and IL-6 contents in the culture supernatant fluid were measured using ELISA. RESULTS: LPS stimulation increased significantly the mRNA and protein expression of TPO and ERK in BV2 cells, especially at the concentration of 1.0 μg/mL for 12 hrs stimulation. There was a significant positive correlation between the mRNA and protein expression of TPO and ERK. CONCLUSIONS: Signal transduction pathway of ERK1/2 participates in the activation of TPO in inflammatory injury of BV2 cells.