Nterleukin-10 Inhibit Interferon-γ Induced The Upregulation of Indoleamine 2,3-Dioxygenase in Astrocytes

【作者】 姜伟； 张金锦； 陈于波； 游自立；

【机构】 电子科技大学生命科学与技术学院；

【摘要】 Indoleamine 2,3-dioxygenase(IDO) is the first and rate-limiting enzyme in catalyzing tryptophan via the kynurenine pathway in mammals.In the condition of inflammation, infection or injury, IDO can be induced to be over expression in central nervous system(CNS), which leads to reduction of tryptophan level and production of series of toxic metabolites.These toxic metabolites make neurons dsyfunction and death.Interleukin-10(IL-10) as a typical anti-inflammatory cytokine not only inhibit inflammatory response, has the function of neuron protection.This study aims to explore the effect and signaling pathway of IL-10 on IDO production and activity in astrocyte.Cortical astrocytes from C57BL/6J neonatal mice were cultured in vitro and treated with 100 μg/ml Interferon-γ(IFN-γ) or/and 10 ng/ml IL-10.IDO mRNA was detected with real-time PCR, and protein expression with immunofluorescence and western blot, IDO activity was assayed using colorimetric method.The results showed that the purity of astrocytes is over 95%.IFN-γ can induced IDO mRNA significantly expression.After 48 h, there is an increasing of IDO positive cells, a high IDO protein level.IL-10 can inhibit IDO expression induced by IFN-γ, which displayed at mRNA, protein level and IDO activities.These findings show that IL-10 can downregulate IDO expression to play the nerve protective role and provide the possibility of anti-inflammatory treatment in regulating tryptophan metabolism in various neurological disorders.更多还原

【Abstract】 Indoleamine 2,3-dioxygenase(IDO) is the first and rate-limiting enzyme in catalyzing tryptophan via the kynurenine pathway in mammals.In the condition of inflammation, infection or injury, IDO can be induced to be over expression in central nervous system(CNS), which leads to reduction of tryptophan level and production of series of toxic metabolites.These toxic metabolites make neurons dsyfunction and death.Interleukin-10(IL-10) as a typical anti-inflammatory cytokine not only inhibit inflammatory response, has the function of neuron protection.This study aims to explore the effect and signaling pathway of IL-10 on IDO production and activity in astrocyte.Cortical astrocytes from C57BL/6J neonatal mice were cultured in vitro and treated with 100 μg/ml Interferon-γ(IFN-γ) or/and 10 ng/ml IL-10.IDO mRNA was detected with real-time PCR, and protein expression with immunofluorescence and western blot, IDO activity was assayed using colorimetric method.The results showed that the purity of astrocytes is over 95%.IFN-γ can induced IDO mRNA significantly expression.After 48 h, there is an increasing of IDO positive cells, a high IDO protein level.IL-10 can inhibit IDO expression induced by IFN-γ, which displayed at mRNA, protein level and IDO activities.These findings show that IL-10 can downregulate IDO expression to play the nerve protective role and provide the possibility of anti-inflammatory treatment in regulating tryptophan metabolism in various neurological disorders.更多还原