目的:优化芦蒿总黄酮(Artemisia selengensis Turcz. total flavonoids,ATTF)提取的工艺条件,并研究其提取物抗炎活性。方法:采用响应面分析法中的中心组合设计方法(central composite design,CCD),建立以复合酶用量、超声温度、乙醇体积分数和液料比对ATTF得率影响的二次回归模型。采用脂多糖(LPS)诱导RAW264. 7巨噬细胞分泌一氧化氮(NO)建立细胞炎症模型,加入不同浓度的ATTF提取物,用NO试剂盒测定NO的分泌量。结果:芦蒿总黄酮的最佳提取工艺条件为复合酶(纤维素酶与果胶酶)用量1.5%、超声温度50 ℃、乙醇体积分数70%、液料比40:1 (mL/g),在此条件下,ATTF的得率为13.09%±2.13%。ATTF提取物中、高剂量组能够明显抑制脂多糖(lipopolysaccharide,LPS)诱导的RAW264.7巨噬细胞NO的分泌。结论:该回归模型有极显著性,可以用来合理预测ATTF的得率;ATTF提取物可以通过抑制巨噬细胞NO的分泌而发挥抗炎作用。
Objective:To optimize the technological conditions of total flavonoids extraction from Artemisia selengensis Turcz.(ATTF)and study the anti-inflammatory activity of the extracts. Methods:A quadratic regression model with the effect of compound enzyme dosage,ultrasonic temperature,ethanol volume fraction and liquid-to-solid ratio on ATTF yield were established using the central composite design(CCD)in response surface analysis. A cell inflammation model was established using lipopolysaccharide(LPS)-induced RAW264.7 macrophages to secrete nitric oxide(NO),and then different concentrations of ATTF extracts were added to determine the secretion of NO with the NO kit. Results:The optimum extraction conditions of total flavonoids from Artemisia selengensis Turcz. were the compound enzyme dosage 1.5%,ultrasonic temperature 50 ℃,ethanol volume fraction 70%,liquid-to-material ratio 40:1 (mL/g). Under these condition,the yield of ATTF was 13.09%±2.13%.The ATTF medium and high doses of the extract could significantly inhibit NO secretion in LPS-induced RAW264.7 macrophages. Conclusion:The regression model was significant,which would be used to reasonably predict the yield of ATTF. ATTF extracts would play an anti-inflammatory role by inhibiting the secretion of NO in macrophages.