盐酸二甲双胍对Aβ_(1-42)诱导的BV-2细胞炎性因子表达的影响及机制

Effect of metformin hydrochloride on the expressions of inflammatory cytokines in Aβ_(1-42)-induced BV-2 cells and its mechanisms

目的研究盐酸二甲双胍对Aβ_(1-42)诱导的BV-2细胞炎性因子表达的影响及机制。方法采用Aβ_(1-42)诱导BV-2细胞构建神经炎症细胞模型,盐酸二甲双胍干预后,MTT法检测细胞活力,ELISA法检测各组细胞上清液中TNF-α、IL-1β、IL-6、IL-18含量,RT-PCR法检测各组细胞TNF-α、IL-1β、IL-6、IL-18 mRNA水平,Western blotting检测NLRP3、ASC、pro-Caspase-1、Caspase-1、p-IκBα、IκBα、NF-κB蛋白的表达。结果1.25、2.5、5μmol/L的Aβ_(1-42)剂量依赖性降低BV-2细胞活力,0.5、1、2 mmol/L盐酸二甲双胍剂量依赖性升高5μmol/L Aβ_(1-42)诱导的BV-2细胞活力、降低TNF-α、IL-1β、IL-6、IL-18的转录及分泌,抑制NLRP3、ASC、Caspase-1(p20)、NF-κB、p-IκBα蛋白表达。结论盐酸二甲双胍可能通过NF-κB通路抑制NLRP3炎性小体的活化,减弱Aβ_(1-42)诱导的BV-2细胞炎症反应。

Objective To study the effect of metformin(Met)hydrochloride on the expressions of inflammatory cytokines in Aβ_(1-42)-induced BV-2 cells and its mechanisms.Methods BV-2 cells were induced by Aβ_(1-42) to mimic the neuroinflammatory cell model,and after treatment with Met,the contents of cytokines,including TNF-α,IL-1β,IL-6 and IL-18,in cell supernatant were examined by ELISA;the mRNA levels of TNF-α,IL-1β,IL-6 and IL-18 were examined by RT-PCR;and the expressions of NLRP3,ASC,pro-Caspase-1,Caspase-1,NF-κB,p-IκBαand IκBαwere examined by Western blotting.Results Aβ_(1-42) at 1.25,2.5 and 5μmol/L could decrease the BV-2 cell viability in a dose-dependent manner,while Met at 0.5,1 and 2 mmol/L could increase the cell viability in 5μmol/L Aβ_(1-42)-induced BV-2 cells.After treatment with Met,the transcription and secretion of TNF-α,IL-1β,IL-6 and IL-18,and the protein expressions of NLRP3,ASC,Caspase-1(p20),NF-κB and p-IκBαwere significantly decreased in Aβ_(1-42)-induced BV-2 cells.Conclusion Met attenuates inflammatory responses in Aβ_(1-42)-induced BV-2 cells,which may be associated with the inhibition of NLRP3 inflammasome activation via NF-κB pathway.