鹰嘴豆芽素A通过PPARα/γ抑制脂多糖诱导猪外周血单个核细胞炎性细胞因子的分泌

Biochanin A suppresses the secretion of proinflammatory cytokines from lipopolysaccharide-stimulated porcine peripheral blood mononuclear cells via PPARα/γ pathways

目的:研究鹰嘴豆芽素A对脂多糖(Lipopolysaccharide,LPS)诱导猪外周血单个核细胞分泌炎性细胞因子的影响。方法:用鹰嘴豆芽素A(50,100μmol/L)处理猪外周血单个核细胞,采用ELISA方法检测其对LPS刺激后细胞分泌肿瘤坏死因子(Tumor necrosis factor-α,TNF-α)、白介素6(Interleukin-6,IL-6)的影响,并用RT-PCR法检测这些细胞因子mRNA的表达,用Western blot法检测核因子κB(Nuclear factorκB,NF-κB)由细胞浆移位至细胞核内情况以判断NF-κB活性变化;同时分别用过氧化物酶体增殖物激活受体(PPAR)α的拮抗剂MK886和PPARγ的拮抗剂GW9662与鹰嘴豆芽素A共同处理猪外周血单个核细胞,观察鹰嘴豆芽素A在PPARα或PPARγ的活性被抑制时对NF-κB活性的影响。结果:鹰嘴豆芽素A能显著降低LPS诱导猪外周血单个核细胞的TNF-α、IL-6的分泌和mRNA的表达;同时鹰嘴豆芽素A显著抑制了LPS诱导猪外周血单个核细胞NF-κB的激活,而此抑制作用均被MK886或GW9662减弱。结论:鹰嘴豆芽素A可通过PPARα和PPARγ抑制LPS诱导猪外周血单个核细胞NF-κB的活化,从而下调LPS诱导的TNF-α、IL-6的生成。

Objective：To investigate the effects of biochanin A on the lipopolysaccharide（LPS） induced secretion of proinflammatory cytokines from porcine peripheral blood mononuclear cells（PBMCs）.Methods：Porcine PBMCs were isolated and pretreated with 50 or 100 μmol/L of biochanin A,the secretion of TNF-α and IL-6 were then determined by ELISA after LPS stimulation,and mRNA expressions were determined by RT-PCR.NF-κB nuclear translocation was detected by Western blot to determine its activity.After suppressing PPARα or PPARγ activity by PPARα antagonist MK886 or PPARγ antagonist GW9662,the effect of biochanin A on LPS-induced NF-κB nuclear translocation were further studied to determine the dependency of biochanin A on PPARα and PPARγ pathway.Results：The LPS-induced secretion and mRNA up-regulation of TNF-α and IL-6 were significantly suppressed in porcine PBMCs by biochanin A;the LPS-stimulated activation of NF-κB was suppressed by biochanin A and the suppression effect was attenuated by MK886 or GW9662.Conclusion：Biochanin A significantly down-regulated TNF-α and IL-6 production in LPS-stimulated porcine PBMCs by inhibiting the activation of NF-κB,and the anti-inflammatory effect of biochanin A was PPARα and PPARγ-dependent.