线纹海马乙醇提取物对脂多糖诱导BV2小胶质细胞炎症反应的作用

Anti-inflammatory and Nitric Oxide(NO) Release Effects of Hippocampus erectus Ethanol Extracts on Lipopolysaccharide-induced BV2 Microglia

【目的】研究线纹海马乙醇提取物对脂多糖(Lipopolysaccharide,LPS)诱导BV2小胶质细胞神经炎症和氧化反应的抑制作用。【方法】利用LPS诱导BV2小胶质细胞建立炎症反应模型,根据对BV2细胞不同的处理方法将其分为对照组、模型组、线纹海马乙醇提取物组(0.1～1 000 ng/mL)和模型组+线纹海马乙醇提取物组(1ng/m L)。用CCK8法检测BV2细胞存活率,显微镜下观察细胞形态的变化,Greiss法测定细胞释放的一氧化NO含量,ELISA法检测BV2细胞上清液中促炎症因子如白细胞介素(interleukin,IL-1β)、IL-6和肿瘤坏死因子(tumornecrosisfactor,TNF-α)的表达量。【结果】与对照组相比,线纹海马乙醇提取物对BV2细胞的增殖无显著影响,而LPS刺激显著促进BV2细胞的增殖,细胞呈典型的阿米巴状,同时大量释放氧自由基NO和促炎细胞因子IL-1β、IL-6和TNF-α。线纹海马乙醇提取物显著抑制LPS诱导的BV2细胞增殖,改善LPS诱导的细胞阿米巴状形态,并抑制LPS激活BV2细胞释放的促炎因子TNF-α、IL-1β、IL-6和氧自由基NO(P <0.05)。【结论】线纹海马乙醇提取物可明显抑制LPS诱导的BV2细胞的增殖、细胞形态的改变及其所产生的炎症因子和氧自由基的增加。

【Objective】To investigate the protective effects of Hippocampus erectus ethanol extracts on the Nitric Oxide(NO)release and inflammatory reaction in BV2 microglia induced by lipopolysaccharide(LPS).【Methods】BV-2 cells were cultured in vitro and LPS was used to construct the inflammatory model in BV-2 cells.The cells were divided into control,model(LPS 100 ng/mL),H.erectus ethanol extracts(0.1~1 000 ng/mL)treated and model(LPS 100 ng/mL)+H.erectus ethanol extracts(1 ng/mL)groups.CCK8 kit was used to determine the proliferation of BV2 microglial,microscope 200 times was used to observe the cellular morphology.Greiss was used to investigate the release of NO and ELISA kit was used to measure the levels of TNF-α,IL-1βand IL-6.【Results】H.erectus ethanol extracts did not show any effects on the survival rate of BV2 microglia when it was compared to the control group.LPS administration significantly increased proinflammatory cytokines,changed the cell morphology into the shape of amoeba,and increased the release of TNF-α,IL-1β,IL-6 as NO.While,H.erectus ethanol extracts significantly inhibited the proliferation rates of BV2 cells induced by LPS(P<0.05),and normalized the cell morphology.Furthermore,Hippocampus erectus ethanol extracts significantly inhibited the levels of TNF-α,IL-1β,IL-6,and NO in LPS-induced BV2 microglia as compared with the model group(P<0.05).【Conclusion】H.erectus ethanol extracts significantly inhibited LPS-induced BV2 microglial proliferation,morphological change,oxidative stress and the inflammatory response.