积雪草苷对脂多糖诱导的RAW264.7细胞核转录因子κB活化及炎症反应的作用

目的探讨积雪草苷对脂多糖(LPS)诱导的RAW264.7巨噬细胞核转录因子κB(NF-κB)活化及炎症因子表达的影响。方法用LPS刺激RAW264.7巨噬细胞建立炎症模型,四甲基偶氮唑盐(MTT)法检测低、中、高浓度积雪草苷(终浓度分别为10-7、10-6及10-5mol/mL)对RAW264.7细胞增殖的影响,激光共聚焦显微镜观察积雪草苷对细胞NF-κB核转运的作用,ELISA法检测细胞上清中细胞因子TNF-α、IL-1及IL-10的变化。结果低、中、高浓度积雪草苷对RAW264.7细胞增殖均无显著影响。积雪草苷明显抑制RAW264.7细胞NF-κB活化,抑制促炎因子TNF-α和IL-1的表达,同时上调抗炎因子IL-10的表达。空白组、模型组及积雪草苷低、中、高浓度干预组NF-κB转入细胞核百分率分别为(3.5±1.5)%、(75.7±9.1)%、(66.8±7.1)%、(58.9±9.0)%、(40.1±8.8)%,细胞上清TNF-α浓度分别为(171.12±35.42、1775.45±193.97、1284.63±162.13、1035.22±187.97、598.90±107.73)pg/mL,IL-1浓度为(5.66±0.98、26.93±3.48、22.41±2.84、17.05±1.70、10.64±1.29)ng/mL,IL-10为(25.23±2.17、71.75±8.31、82.82±6.00、98.70±8.84、119.97±9.13)pg/mL。结论积雪草苷可能通过抑制NF-κB信号通路,维持促炎系统与抗炎系统的平衡而发挥抗炎作用。

Effects of Asiaticoside On Activation of Nuclear Factor kappa B and Inflammation in RAW264.7 Cells Induced by Lipopolysaccharide

Objective To explore the effects of asiaticoside on the activation of nuclear factor kappa B （NF-κB） and cytokines expression in RAW264.7 cells induced by lipopolysaccharide （LPS）.Methods RAW264.7 cells were allocated to 5 groups,ie.a blank group,a model group stimulated by LPS at dose of 10 μg/mL,and three asiaticoside treatment groups stimulated by LPS and different doses of asiaticoside simultaneously.The effects of asiaticoside（10-7,10-6,10-5 mol/mL） on the proliferation of cells were examined by MTT assay.The activation of NF-κB was detected and analyzed by the laser scanning confocal microscope（LSCM）,meanwhile the concentrations of TNF-α,IL-1,and IL-10 in supernatants were quantified by ELISA.Results MTT assay showed that asiaticoside （10-7,10-6,10-5 mol/mL） had no effects on the proliferation of RAW264.7 cells.Asiaticoside significantly decreased the activation of NF-κB,downregulated the secretion of TNF-α and IL-1,and upregulated IL-10 secretion in a dose dependent manner.According to LSCM,the ratio of NF-κB activation was （3.5±1.5）%,（75.7±9.1）%,（66.8±7.1）%,（58.9±9.0）%,and （40.1±8.8）% in the blank,model,and asiaticoside（10-7,10-6,10-5 mol/mL） treatment groups respectively.The contents of TNF-α in supernatants were （171.12±35.42,1775.45±193.97,1284.63±162.13,1035.22±187.97,598.90±107.73）pg/mL respectively and IL-1 were （5.66±0.98,26.93±3.48,22.41±2.84,17.05±1.70,10.64±1.29）ng/mL respectively,while IL-10 were （25.23±2.17,71.75±8.31,82.82±6.00,98.70±8.84,119.97±9.13）pg/mL respectively.Conclusion The antiinflammation mechanism of asiaticoside may be mediated by downregulating inflammatory factors through NF-κB signal pathway and keeping the balance between proinflammatory and antiinflammatory system.