朝藿定B对BV2小胶质细胞炎症反应的作用及机制

ROLE AND MECHANISM OF EPIMEDIN B IN INFLAMMATORY RESPONSE IN BV2 MICROGLIA CELLS

目的探讨朝藿定B对脂多糖(LPS)诱导的BV2小胶质细胞炎症反应的抑制作用及雌激素受体(ER)特异性阻断剂ICI 182,780的阻断效应。方法常规培养BV2小胶质细胞,加或不加朝藿定B、LPS、ICI 182,780处理,采用3-(4,5-二甲基-2-噻唑基)-2,5二苯基溴化四唑(MTT)比色法检测细胞活力,实时聚合酶链反应检测肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)mRNA的表达。结果不同浓度朝藿定B对BV2小胶质细胞活力没有影响(P>0.05)。LPS可明显上调促炎因子TNF-α(F=10.64,q=9.157,P<0.001)和IL-6(F=25.25,q=12.630,P<0.001)mRNA的表达;1、10μmol/L朝藿定B预处理均能明显抑制LPS诱导的TNF-αmRNA表达上调(q=4.655、5.408,P<0.05),10μmol/L朝藿定B对LPS诱导的IL-6 mRNA表达有明显的抑制作用(q=4.696,P<0.05)。朝藿定B对LPS诱导的TNF-α和IL-6 mRNA表达的抑制作用可被ICI 182,780所阻断(F=12.53、28.71,q=5.318、4.684,P<0.05)。结论朝藿定B能够抑制LPS诱导的BV2小胶质细胞促炎因子的表达,其机制可能与ER信号途径有关。

Objective To investigate the inhibitory effect of Epimedin B on lipopolysaccharide(LPS)-induced inflammatory response in BV2 microglial cells and the blocking effect of the estrogen receptor(ER)-specific blocker ICI 182,780.Methods BV2 microglial cells were routinely cultured and treated with or without Epimedin B,LPS,and ICI 182,780.MTT colorimetry was used to measure cell viability,and real-time PCR was used to measure the mRNA expression levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6).Results Different concentrations of Epimedin B had no effect on the viability of BV2 microglial cells(P>0.05).LPS significantly upregulated the mRNA expression levels of the proinflammatory factors TNF-α(F=10.64,q=9.157,P<0.001)and IL-6(F=25.25,q=12.630,P<0.001).Epimedin B pretreatment at concentrations of 1 and 10μmol/L significantly inhibited the upregulated mRNA expression of TNF-αinduced by LPS(q=4.655,5.408;P<0.05),and Epimedin B at a concentration of 10μmol/L significantly inhibited the LPS-induced mRNA expression of IL-6(q=4.696,P<0.05).The inhibitory effect of Epimedin B on the LPS-induced mRNA expression of TNF-αand IL-6 could be blocked by ICI 182,780(F=12.53,28.71;q=5.318,4.684;P<0.05).Conclusion Epimedin B can inhibit the expression of proinflammatory factors induced by LPS in BV2 microglial cells,which may be associated with the ER signaling pathway.