紫铆因抑制脂多糖诱导小胶质细胞炎性反应的作用机制研究

Study on mechanism of butein on inhibiting LPS induced microglia inflammatory reaction

目的探讨紫铆因抑制脂多糖(LPS)诱导BV2小胶质细胞炎性反应的作用机制。方法以LPS诱导BV2小胶质细胞活化建立体外神经炎症细胞模型。MTT法检测紫铆因对BV2细胞存活率的影响;qPCR法检测各组细胞中炎症因子白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)的mRNA表达水平;Western blot法检测各组细胞内核因子-κB(NF-κB)p65和ERK信号通路相关蛋白的表达变化;免疫荧光染色观察各组细胞内NF-κB p65的核转录活性变化。结果紫铆因干预BV2细胞24h后,对细胞活力无明显影响;紫铆因能显著下调LPS诱导BV2细胞活化后炎症因子IL-1β、IL-6和TNF-α的mRNA表达水平;紫铆因能明显降低LPS诱导BV2细胞活化后,ERK信号通路相关蛋白的磷酸化水平,并且能有效抑制NF-κB p65的核转录活性,阻止其向核内转移。结论紫铆因可以抑制LPS诱导的小胶质细胞活化,降低BV2小胶质细胞的炎性反应。

Objective To investigate the mechanism of butein on inhibiting LPS induced BV2 microglia inflammatory reaction.Methods LPS induced BV2 microglia activation was used to establish the in vitro neuroinflammation cell model.The MTT assay was used to detected the effects of butein on BV2 cells survival rate;the inflammatory factor IL-1β,IL-6 and TNF-α mRNA expression levels were measured by qPCR;the NF-κB p65 protein expression and ERK signaling pathway related protein expression changes were detected by Western blot;NF-κB p65 nuclear transcriptional activity in each group was detected by immunofluorescence staining.Results Butein intervention on BV2 cells for 24 h,had no effect on cell viability;butein could significantly inhibit IL-1β,IL-6 and TNF-αmRNA expression level after LPS inducing BV2 cell activation;butein could significantly reduce ERK signaling pathway related proteins phosphorylation level after LPS inducing BV2 cells activation,moreover could effectively inhibit the NF-κB p65 nuclear transcriptional activity,and prevent it transfer to intranuclear.Conclusion Butein can inhibit LPS induced BV2 microglia activation and reduces the inflammatory reaction of microglias.