摘要
目的:研究天麻素(GAS)对脂多糖(LPS)激活的BV2小胶质细胞性别决定区Y框蛋白2(SOX2)/β-连环蛋白(β-catenin)通路表达的影响.方法:体外培养BV2小胶质细胞,分为对照组(Control)、LPS刺激组(LPS)、LPS+0.17 mmol/L GAS 处理组(LPS+GAS-L)、LPS+0.34 mmol/L GAS 处理组(LPS+GAS-H)、SOX2抑制剂丙萘洛尔(PR)处理组(PR)、LPS+PR处理组(LPS+PR)、LPS+PR+GAS处理组(LPS+PR+GAS).通过CCK-8检测PR对BV2小胶质细胞活力的影响,并通过Western Blot和免疫荧光双标染色检测SOX2、β-cate-nin、甘露糖受体(CD206)和肿瘤坏死因子-α(TNF-α)的表达变化.结果:PR在0～40 µmol/L范围内不会引起显著的BV2小胶质细胞死亡;LPS刺激组中SOX2、β-catenin和TNF-α蛋白表达明显增强,而CD206表达显著降低(P＜0.05).GAS干预后SOX2、β-catenin和TNF-α表达显著减少,而CD206明显升高(P＜0.05);与LPS组相比,用PR阻断SOX2后β-catenin和TNF-α蛋白表达显著降低(P＜0.05),而PR与GAS联合应用与单纯GAS干预组相比,β-catenin和TNF-α蛋白表达无明显差异.结论:天麻素可能通过抑制SOX2/β-catenin通路减轻小胶质细胞的激活,发挥抗炎作用.
Abstract
Objective:To investigate the effect of gastrodin(GAS)on the sex-determining region Y-box2(SOX2)/β-catenin pathway in microglia induced by lipopolysaccharide(LPS).Methods:BV2 microglia was cultured in vitro and divided into the following groups:Control group(Control),LPS group(LPS),LPS+0.17 mmol/L gastrodin treatment group(LPS+GAS-L),LPS+0.34 mmol/L gastrodin treatment group(LPS+GAS-H),SOX2 inhibitor pronethalolgroup(PR),LPS+PR group(LPS+PR),and LPS+PR+GAS group(LPS+PR+GAS).Effect of PR on BV2 microglia viability was detected by CCK-8.The expression of SOX2,β-catenin,mannose receptor(CD206)and tumor necrosis factor-α(TNF-α)was assessed using Western Blot and immunofluorescence double staining.Results:PR did not induce significant BV2 cell death in the 0～40 μmol/L range.After LPS treatment,the expression levels of SOX2,β-catenin,and TNF-α significantly increased in the LPS group,while CD206 decreased(P＜0.05).Following GAS treatment,the expression levels of SOX2,β-catenin,and TNF-α significantly decreased,while CD206 increased(P＜0.05).Compared to the LPS group,the expression levels of β-catenin and TNF-α significantly de-creased in the PR group(P＜0.05),but no significant difference was observed between the LPS+GAS and LPS+PR+GAS group.Conclusion:GAS significantly inhibits LPS-induced microglia activation potentially through the inhibi-tion of the SOX2/β-catenin signaling pathway,and exerts anti-inflammatory effects.
基金项目
国家自然科学基金(31960194)
云南省大学生创新性实验计划项目(2022JXD273)
NETL
NSTL
万方数据