细胞型朊蛋白在BV2小神经胶质细胞体外激活中的作用

Effects of Prion Protein on the Regulation of Classical and Alternative Activation of BV2 Microglia in vitro

【目的】研究细胞型朊蛋白对小神经胶质细胞不同激活方式的影响。【方法】用IFN-γ、IL-4和IL-10分别刺激BV2细胞,RT-PCR方法检测PrPC的mRNA表达量。SiRNA干扰将PrPC沉默,用上述因子刺激细胞,用RT-PCR和Western-blot检测相关参数。【结果】用IFN-γ、IL-4和IL-10分别刺激小神经胶质细胞后可导致PrPC的mRNA表达量下降;PrPC沉默后的小神经胶质细胞对IFN-γ刺激的反应应答减弱;PrPC沉默可以显著地改变由IL-4诱导的神经胶质细胞的激活表型;但是对IL-10诱导的小神经胶质细胞激活却没有影响。【结论】PrPC既能影响小神经胶质细胞从静止状态到激活状态的转换,也在小神经胶质细胞的经典激活和替代激活途径中发挥调节作用。

[ Objective ] The purpose of this study is to investigate the effects of PrPc on various forms of microglial activation. [Method] BV2 microglia were treated, respectively, with IFN-γ, IL-4, or IL-10, and the mRNA expression of PRNP was examined by RT-PCR. Then the effects of si-RNA-mediated disruption of PRNP on different parameters of microglial activation in IFN-γ, IL-4, or IL-10-stimulated microglia were analyzed by RT-PCR and western-blot. [Result] PRNP mRNA expression was invariably downregulated in microglia upon exposure to IFN-y, IL-4, or IL-10. PRNP silencing prior to cytokines treatment reduced the responsiveness of microglia to INF-γ treatment, significantly altered IL-4-induced microglial activation phenotype, and had no effect on IL-10-induced microglial activation. [Conclusion] Together, these results support a role ofPrPc in the modulation of the shift of microglia from a quiescent state to an activated phenotype and in the regulation of the mieroglial response during classical and alternative activation.