凝血酶诱导培养星形胶质细胞炎症细胞因子的表达

Thrombin induces the expression of cytokine in cultured astrocytes

目的证实凝血酶诱导体外培养的大鼠星形胶质细胞(astrocyte,As)白介素-1β(inter-leukin-1 beta,IL-1β)和干扰素-γ(Interferon-gamma,IFN-γ)的表达。方法采用体外培养As的方法,对新生Wistar大鼠大脑As进行培养并纯化,用不同浓度的凝血酶干预;酶联免疫吸附法(en-zyme-linked immunosorbent assay,ELISA)检测培养大鼠脑皮层As上清液中IL-1β、γ-干扰素的含量,免疫细胞化学染色和逆转录多聚酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)检测培养AsIL-1β、IFN-γ蛋白和基因的表达。结果凝血酶上调体外培养As上调IL-1β、IFN-γ蛋白及mRNA的表达。结论凝血酶诱导培养As上细胞因子的IL-1β、IFN-γ表达,这些炎症细胞因子有助于As的损伤。

Objective To determine thrombin up-regulate the expression of IL-1β and IFN-γ in cultured cortical astrocytes, and the effects on cell proliferation. Methods Primary astrocytes were dissociated from new-born wistar rats and cultured to purify them. The purified astrocytes were treated by thrombin at different concentration. Enzyme- linked immunosorbent assay was applied to examine the level of IL-1β and IFN-γ in cultured supernatant; Immunocytochemistry staining and RT-PCR were used to measure the protein and gene expression of IL-1β and IFN-γ in cultured cortical astrocytes. Results The protein and mRNA level of IL-1β and IFN-γ in cultured cortical astrocytes were upregu- lated by thrombin, thrombin made the viability of astrocytes decreased. Conclusions The expression of IL- 1β and IFN-γ in cultured cortical astrocytes were induced by thrombin, which could associate to the astrocytes damage.