加味五子衍宗方及其活性部位对脂多糖诱导神经炎症反应的抑制作用及机制研究

Inhibitory effects of modified Wuzi-Yanzong prescription and its active components on LPS-induced inflammatory response on BV-2 microglial cells

目的研究加味五子衍宗方及其活性部位对脂多糖诱导的BV-2小胶质细胞炎症反应的抑制作用及潜在机制。方法通过大孔树脂柱,将加味五子衍宗方乙醇总提取物洗脱分离得到水提组、20%醇提组、50%醇提组、70%醇提组和95%醇提组5个部位。针对无细胞毒性的部位研究了其对炎症因子一氧化氮和炎症蛋白诱导型一氧化氮合酶,环氧合酶-2表达的影响,明确了抗炎活性部位,同时进一步研究了活性部位对核转录因子的激活以及活性氧表达的调控作用。结果70%醇提组和95%醇提组对细胞具有毒性,其余各部位(水提组,20%醇提组,50%醇提组)均无细胞毒性且表现出一定的抗炎活性。其中50%醇提组抗炎活性最优,甚至高于复方组。结论加味五子衍宗方的抗炎活性部位可能主要富集于50%醇提部分,其抗炎活性可能是通过抑制NF-κB信号通路介导的炎症反应以及抗氧化应激实现的。

Objective To study the inhibitory effects of modified Wuzi-Yanzong prescription （ MWP ） and its active components on LPS-induced BV-2 microglial cells and the potential mechanism. Methods 70% ethanol extracts of MWP was separated by macro porous resin column, collecting water elution group, 20% alcohol elution group, 50% alcohol elution group, 70% alcohol elution group and 95% alcohol elution group. For the non-cytotoxic components, we detected the expression of nitric oxide, inducible nitric oxide synthase and cyclooxygenase-2 to identify the anti-inflammatory active components. The regulatory effects of active components on nuclear factor κB （ NF-κB ） activation and expression of reactive oxygen species（ ROS） were also investigated. Results 70% ethanol and 95% ethanol elution components were toxic to cells while the remaining components （ water elution component,20% alcohol elution component,50% alcohol elution component） didn’t show any cytotoxicity and exhibited some anti-inflammatory activity. The 50% alcohol elution component is better than the other components or even higher than MWP. Conclusions 50% alcohol extracts are responsible for the major anti-inflammatorynbsp;effective component of MWP and the anti-inflammatory activity may be achieved through inhibiting NF-κB signaling pathway and oxidative stress.