基于RAW264.7抗炎细胞模型对红活麻活性部位的筛选及其化学成分研究
|
摘要
目的研究红活麻的抗炎活性部位及其化学成分。方法萃取法制备不同极性部位,采用脂多糖(LPS)刺激小鼠巨噬细胞RAW264.7模型,检测红活麻不同提取部位对NO释放量的影响,考察其抗炎活性;采用硅胶柱色谱和ODS柱色谱技术对红活麻抗炎活性部位的化学成分进行分离纯化,运用波谱分析技术鉴定化合物结构。结果利用SAS 9.30软件对给药组和LPS组NO的含量进行t检验,比较组间是否存在差异;与LPS组相比,石油醚部位对NO的释放量影响不明显(P<0.05)。二氯甲烷部位、乙酸乙酯部位和正丁醇部位在浓度为15.5、31.25、62.5μg/m L时,均表现出对NO释放量有一定的影响,且差异具有统计学意义(P>0.05)。当给药浓度都在62.5μg/m L时,石油醚部位、二氯甲烷部位、乙酸乙酯部位和正丁醇部位分别对NO释放量的抑制为11.42%、21.01%、33%、26.96%,故乙酸乙酯部位对NO释放量影响最大,且具有剂量依赖性;从红活麻乙酸乙酯部位中分离得到7个化合物,分别鉴定为:(1)!-Sitosterol;(2)5-Hydroxymethyl-2-furancarboxaldehyde;(3)4-hydroxy-5-methoxy-benzoic acid;(4)Isomeranzin;(5)Dioctyl phthalate;(6)Dibutyl phthalate;(7)Caffeic acid。结论乙酸乙酯部位为红活麻抗炎活性部位。2,4,5,6为首次从该属中分离得到,7为首次从该植物中分离得到。
Objective To screen the active fraction with anti-inflammatory effect of urtica dentata handand and to study the chemical constituents of the active fraction. Methods Different polar fractions were prepared by extraction with organic solvents. The anti-inflammatory effects were observed by a in vitro model of LPS-stimulated RAW264. 7 macrophages. Chemical constituents of the active fraction extract from urtica dentata hand was purified by silica gel and ODS chromato-graphic columns,and the compounds structures were identified by using spectral analysis techniques. Results Compared the differences between the groups,using SAS9. 30 software to test the content of NO in the administration group and the LPS group by student's t test. The effect of petroleum ether on the release of NO was not obvious,and the difference was not statistically significant. For the methylene chloride site,when the concentration was 15. 5,31. 25 and 62. 5μg/m L in the site of dichloromethane,ethyl acetate and n-butanol,there was a certain effect on the amount of NO release,and the difference was statistically significant. When the concentration of the drug reaches 62. 5 μg/m L,the inhibition rate of the oil ether,dichloromethane,ethyl acetate and positilitol is 11. 42%,21. 01%,33%,26. 96%. Therefore,ethyl acetate was the most significant and dose dependent. Seven compounds were isolated in ethyl acetate extract from urtica dentata hand,They were identified as-sitosterol,bis( 5-formylfurfuryl) ether,4-hydroxy-5-methoxy-benzoic acid,isomeranzin,dioctyl phthalate,dibutyl phthalate,caffeic acid. Conclusion The ethyl acetate extract was the anti-inflammatory fraction of urtica dentata hand. 2,4,5,6 for the first time from the genus,7 was isolated from the plant for the first time.
Objective To screen the active fraction with anti-inflammatory effect of urtica dentata handand and to study the chemical constituents of the active fraction. Methods Different polar fractions were prepared by extraction with organic solvents. The anti-inflammatory effects were observed by a in vitro model of LPS-stimulated RAW264. 7 macrophages. Chemical constituents of the active fraction extract from urtica dentata hand was purified by silica gel and ODS chromato-graphic columns,and the compounds structures were identified by using spectral analysis techniques. Results Compared the differences between the groups,using SAS9. 30 software to test the content of NO in the administration group and the LPS group by student's t test. The effect of petroleum ether on the release of NO was not obvious,and the difference was not statistically significant. For the methylene chloride site,when the concentration was 15. 5,31. 25 and 62. 5μg/m L in the site of dichloromethane,ethyl acetate and n-butanol,there was a certain effect on the amount of NO release,and the difference was statistically significant. When the concentration of the drug reaches 62. 5 μg/m L,the inhibition rate of the oil ether,dichloromethane,ethyl acetate and positilitol is 11. 42%,21. 01%,33%,26. 96%. Therefore,ethyl acetate was the most significant and dose dependent. Seven compounds were isolated in ethyl acetate extract from urtica dentata hand,They were identified as-sitosterol,bis( 5-formylfurfuryl) ether,4-hydroxy-5-methoxy-benzoic acid,isomeranzin,dioctyl phthalate,dibutyl phthalate,caffeic acid. Conclusion The ethyl acetate extract was the anti-inflammatory fraction of urtica dentata hand. 2,4,5,6 for the first time from the genus,7 was isolated from the plant for the first time.
引文
[1]贵州省药品监督管理局.贵州省中药材、民族药材质量标准[S].贵阳:贵州科技出版社,2003:187.
[2]国家中医药管理局《中华本草》编委会.中华本草·苗药卷[M].贵阳:贵州科技出版社,2005:283-284.
[3]罗迎春,孙庆文.贵州民族常用天然药物[M].2卷.贵阳:贵州科技出版社,2013:231-232.
[4]Aydin T,Cakir A,Kazaz C,et al.Insecticidal metabolites from the rhizomes of Veratrum album against adults of Colorado potato beetle,Leptinotarsa decemlineata[J].Chemistry&Biodiversity,2015,11(8):1192-1204.
[5]陈云飞,刘守金,王飞.阔带凤丫蕨倍半萜类化学成分研究[J].中国中药杂志,2012,37(7):946-950.
[6]Gutierrezlugo MT,Woldemicheol GM,Singh MP,et al.Isolation of three new naturally occurring compounds from the culture of Micromonospora sp.P1068[J].Natural Product Research,2015,19(7):645-652.
[7]牛艳,王磊,黄晓君,等.化橘红香豆素类的化学成分[J].暨南大学学报,2012,33(5):501-505.
[8]Essam,Abdel-Sattar,Sahar,et al.New sulphide derivative from Ferula rutabensis[J].Natural Product Research,2009,23(9):861-865.
[9]魏华,何春年,彭勇,等.川木香化学成分研究[J].中国中药杂志,2012,37(9):1249-1253.
[10]田富饶,张琳,田景奎,等.南刘寄奴的化学成分研究[J].中国药物化学杂志,2008,18(5):362-365.
[2]国家中医药管理局《中华本草》编委会.中华本草·苗药卷[M].贵阳:贵州科技出版社,2005:283-284.
[3]罗迎春,孙庆文.贵州民族常用天然药物[M].2卷.贵阳:贵州科技出版社,2013:231-232.
[4]Aydin T,Cakir A,Kazaz C,et al.Insecticidal metabolites from the rhizomes of Veratrum album against adults of Colorado potato beetle,Leptinotarsa decemlineata[J].Chemistry&Biodiversity,2015,11(8):1192-1204.
[5]陈云飞,刘守金,王飞.阔带凤丫蕨倍半萜类化学成分研究[J].中国中药杂志,2012,37(7):946-950.
[6]Gutierrezlugo MT,Woldemicheol GM,Singh MP,et al.Isolation of three new naturally occurring compounds from the culture of Micromonospora sp.P1068[J].Natural Product Research,2015,19(7):645-652.
[7]牛艳,王磊,黄晓君,等.化橘红香豆素类的化学成分[J].暨南大学学报,2012,33(5):501-505.
[8]Essam,Abdel-Sattar,Sahar,et al.New sulphide derivative from Ferula rutabensis[J].Natural Product Research,2009,23(9):861-865.
[9]魏华,何春年,彭勇,等.川木香化学成分研究[J].中国中药杂志,2012,37(9):1249-1253.
[10]田富饶,张琳,田景奎,等.南刘寄奴的化学成分研究[J].中国药物化学杂志,2008,18(5):362-365.