黑蒜多糖的斑马鱼体内抗氧化、抗炎症活性分析

Analysis of Anti-oxidant and Anti-inflammatory Activities of Black Garlic Polysaccharides in Zebrafish in Vivo

采用Plackett-Burman及响应面试验优化黑蒜多糖的提取工艺,确定其最优提取条件为:提取2次、固液比30倍、提取时间2 h、乙醇3倍、提取温度70℃、超声功率300 W、醇沉时间16 h、超声时间10 min,在该条件下提取率达3.65%。体外抗氧化试验证明,黑蒜多糖具有较强的清除自由基能力(羟基自由基、DPPH自由基、ABTS自由基)以及良好的还原力和总抗氧化能力。用甲硝锉建立斑马鱼氧化应激模型,结果表明,黑蒜多糖通过清除自由基发挥显著的抗氧化作用(P<0.01)。通过建立脂多糖(LPS)诱导的斑马鱼炎症模型发现,黑蒜多糖可显著抑制炎症导致的活性氧簇(ROS)形成(P<0.01)。通过建立硫酸铜诱导的斑马鱼炎症模型发现,在黑蒜多糖的干预下中性粒细胞向侧线的迁移数明显减少(P<0.01)。抗炎试验表明,黑蒜多糖具有良好的抗炎能力,可通过抑制ROS信号的释放减轻炎症反应。

Plackett-Burman and response surface experimental design methods were used to optimize the extraction process of black garlic polysaccharides.The optimized extraction process parameters were as follows:extraction times was 2 times,solid-liquid ratio was 30 times,extraction time was 2 h,ethanol multiple was 3 times,extraction temperature was 70℃,ultrasonic power was 300 W,alcohol precipitation time was 16 h,ultrasonic time of 10 min.Under these conditions,the extraction rate of black garlic polysaccharides can reach 3.65%.The results of in vitro antioxidant experiments showed that black garlic polysaccharides have a strong ability to scavenge free radicals(hydroxyl radicals,DPPH radicals,ABTS radicals)as well as good reducing power and total antioxidant capacity.A zebrafish oxidative stress model was established using metronidazole.The results showed that black garlic polysaccharides can exert significant anti-oxidant effects through scavenging free radicals(P<0.1).The zebrafish inflammation model was established using lipopolysaccharide(LPS).The results showed that black garlic polysaccharides can significantly inhibit the formation of reactive oxygen species(ROS)caused by inflammation(P<0.1).By establishing a copper sulfate-induced zebrafish inflammation model,it was found that the number of migrating neutrophils to the lateral line under the intervention of black garlic polysaccharides was significantly reduced(P<0.1).Anti-inflammatory experiments have shown that black garlic has good anti-inflammatory ability and can reduce inflammation by inhibiting the release of ROS signaling.