乳香醋炙前后13种乳香酸成分含量变化及活性比较研究  

Comparative study on changes and activity of 13 boswellic acids in Olibanum before and after vinegar-processed

作　　者：董运茁 张弋[1] 刘振丽[2] 王淳[2] 彭诗涛 万晓莹 宋志前[2] 宁张弛 DONG Yun-zhuo;ZHANG Yi;LIU Zhen-li;WANG Chun;PENG Shi-tao;WAN Xiao-ying;SONG Zhi-qian;NING Zhang-chi(Department of Pharmacy,Tianjin First Central Hospital,Tianjin 300192,China;Institute of Basic Theory,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]天津市第一中心医院药学部,天津300192 [2]中国中医科学院中医基础理论研究所,北京100700

出　　处：《中草药》2021年第23期7128-7137,共10页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(81873009);国家自然科学基金项目(82003950);中国中医科学院中医基础理论研究所中央级科研院所自主选题(YZ-202023);天津市第一中心医院科技基金项目(院2020CF13);中国中医科学院优秀青年科技人才(创新类)基金(ZZ14-YQ-035);中国中医科学院科技创新工程项目(CI2021A04201)。

摘　　要：目的通过对比乳香醋炙前后11-羰基-β-乳香酸(11-keto-boswellic,KBA)、3-乙酰-11-羰基-β-乳香酸(3-acetyl-11-keto-boswellic acid,AKBA)、榄香醇酸、β-榄香酮酸、tsugaric acid A、9,11-去氢-α-乳香酸、9,11-去氢-β-乳香酸、α-乳香酸、β-乳香酸、3-乙酰-9,11-去氢-α-乳香酸、3-乙酰-9,11-去氢-β-乳香酸、3-乙酰α-乳香酸(3-acetyl-α-boswellic acid,α-ABA)和3-乙酰β-乳香酸(3-acetyl-β-boswellic acid,β-ABA)13个乳香酸成分含量变化,探究不同炮制条件对含量变化的影响,考察差异乳香酸成分抗炎活性,初步揭示乳香醋炙增效机制。方法制备不同炮制温度和炮制时间醋乳香,基于超高效液相色谱仪串联三重四级杆质谱仪(UPLC-TQ-MS)建立13种乳香酸成分含量测定方法。采用Acquity UPLC BEH C18色谱柱(100mm×2.1 mm,1.7μm),以0.1%甲酸+5 mmol/L乙酸铵水溶液-0.1%甲酸乙腈溶液为流动相,梯度洗脱,体积流量0.3 mL/min,柱温40℃,质谱采用电喷雾负离子源,多反应监测(multi-reaction monitoring,MRM)模式;建立脂多糖诱导巨噬细胞的炎症细胞模型,通过ELISA法测定乳香、醋乳香、单体(3-乙酰-9,11-去氢-β-乳香酸)和乳香+单体(3-乙酰-9,11-去氢-β-乳香酸)对炎症细胞因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)以及IL-6的影响。结果随醋炙温度升高或炮制时间的延长,乳香中榄香醇酸、β-榄香酮酸、tsugaric acid A、9,11-去氢-α-乳香酸、9,11-去氢-β-乳香酸、3-乙酰-9,11-去氢-α-乳香酸、3-乙酰-9,11-去氢-β-乳香酸、α-ABA和β-ABA 9个乳香酸含量升高幅度增大,而KBA、AKBA、α-乳香酸、β-乳香酸4个成分含量降低幅度增大,其中3-乙酰-9,11-去氢-β-乳香酸含量变化幅度最大。炮制温度引起的成分变化强于炮制时间的影响。乳香酸类成分含量升高或降低与成分结构有关;乳香醋炙后抑制炎症因子分泌的作用显著增强,3-乙酰-9,11-�Objective To establish a UPLC-TQ-MS method for the simultaneous determination of 13 boswellic acids included 11-keto-boswellic(KBA), 3-acetyl-11-keto-boswellic acid(AKBA), elemolic acid, β-elemonic acid, tsugaric acid A, 9,11-dehydro-α-boswellic acid, 9,11-dehydro-β-boswellic acid, α-boswellic acid, β-boswellic acid, 3-acetyl-9,11-dehydro-α-boswellic acid, 3-acetyl-9,11-dehydro-β-boswellic acid, 3-acetyl-α-boswellic acid(α-ABA) and 3-acetyl-β-boswellic acid(β-ABA) in raw Olibanum and vinegar-processed Olibanum, compare the effect of different temperatures and times on the content of 13 index components and reveal the anti-inflammation activity of differential boswellic acids. Methods The Olibanum were processed at different temperatures and times. The ultra-performance liquid chromatograph-tandem triple-quadrupole mass spectrometer(UPLC-TQ-MS) method was established for the simultaneous determination of 13 boswellic acids. Acquity UPLC BEH C18 column(100 mm × 2.1 mm, 1.7 μm)was eluted with the mobile phase of water(containing 0.1% formic acid and 5 mmol/L ammonium acetate) and acetonitrile(containing 0.1% formic acid) with gradient elution. The flow rate was 0.3 mL/min. The column temperature was maintained at 40 ℃. The negative ion mode was chosen. Multiple reaction monitoring(MRM) was employed for quantification;The cells model was induced by lipopolysaccharide, the levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) and interleukin-6(IL-6) after the treatment of Olibanum, vinegar-processed Olibanum, 3-acetyl-9,11-dehydro-β-boswellic acid and Olibanum + compound(3-acetyl-9,11-dehydro-β-boswellic acid) were determined by ELISA method. Results With the increase of temperature and time, the content of nine compounds in vinegar-processed frankincense, including elemolic acid, β-elemonic acid, tsugaric acid A, 9,11-dehydro-α-boswellic acid, 9,11-dehydro-β-boswellic acid, 3-acetyl-9,11-dehydro-α-boswellic acid, 3-acetyl-9, 11-dehydro-β-boswellic acid, α-ABA and β-ABA were incre

关 键 词：乳香 醋炙 乳香酸 抗炎 UPLC-TQ-MS ELISA法 11-羰基-β-乳香酸 3-乙酰-11-羰基-β-乳香酸 榄香醇酸榄香醇酸 β-榄香酮酸 tsugaric acid A 9 11-去氢-α-乳香酸 9 11-去氢-β-乳香酸 α-乳香酸 β-乳香酸 3-乙酰-9 11-去氢-α-乳香酸 3-乙酰-9 11-去氢-β-乳香酸 3-乙酰α-乳香酸 3-乙酰β-乳香酸乳香酸 

分 类 号：R283.1[医药卫生—中药学]