摘要
目的研究紫花地丁乙醇回旋冷凝法分离乙酸乙酯提取物和水提取物的化学成分及抗炎活性。方法紫花地丁乙醇回旋冷凝法提取乙酸乙酯部位和水相部位,并对各部位进行化学成分检识。采用二甲苯诱导小鼠耳廓肿胀、小鼠腹腔毛细血管通透性、角叉菜胶诱导大鼠足趾肿胀进行急性炎症造模、炎性组织PGE2含量检测、NO含量检测等动物试验,研究紫花地丁不同提取物的抗炎作用。结果紫花地丁乙酸乙酯提取物中主要含有黄酮、蒽醌类,水提取物中主要含有糖及苷类、皂苷类。乙酸乙酯提物各剂量组对二甲苯致小鼠耳肿胀、角叉菜胶致小鼠足肿胀、小鼠腹腔毛细血管通透性均具有显著的抑制作用(P<0.05~P<0.01),并可不同程度降低炎性组织PGE2含量和NO含量,中剂量组作用极显著(P<0.01)。结论紫花地丁的抗炎作用活性成分在乙醇回旋冷凝法提取的乙酸乙酯提取物中;紫花地丁乙酸乙酯提取物抑制了炎性组织中PGE2的生成量和NO释放量,其抗炎作用可能与降低PGE2、NO表达有关。
Objective To study the anti-inflammatory activity of ethyl acetate extract and water extract from Viola yedoensis by ethanol cyclotron condensation. Methods Ethyl acetate and water phase fractions were extracted by ethanol cyclotron condensation from Violet Flower Dibutylethanol,and the chemical constituents of each fraction were identified.In order to study the anti-inflammatory effects of different extracts of Radix Zihua Ding,xylene-induced ear swelling,abdominal capillary permeability,carrageenan-induced toe swelling in rats were used to establish acute inflammation model,detect the content of PGE2 in inflammatory tissues,and detect the content of NO. Results Flavonoids and anthraquinones are the main components of ethyl acetate extract of Viola florida,and sugar,glycosides and saponins are the main components of water extract.Ethyl acetate extract had significant inhibitory effects on xylene-induced ear swelling,carrageenan-induced foot swelling and abdominal capillary permeability in mice(P<0.05-P <0.01),and could reduce the contents of PGE2 and NO in inflammatory tissues to varying degrees.The middle dose group had significant inhibitory effects(P<0.01). Conclusion Ethyl acetate extract of Viola Florida has significant anti-inflammatory effect,and its anti-inflammatory mechanism may be related to the decrease of PGE2 and NO expression.
引文
[1]中华人民共和国国家药典委员会.中国药典[M].北京:中国医药科技出版社,2015:337.
[2]Zhu H,Qin S S,Zhang N,et al.Chemical constituents and biological activities of plants from the genus viola[J].Chem Biodiver,2015,12(12):1777-1808.
[3]Du D S,Cheng Z H,Chen D F.Anticomplement sesqui terpenes from viola yedoensis[J].Fitoterapia,2015,101:73-79.
[4]Pan Y Y,Song Z P,Zhu G F,et al.Antipyretic effects of liposoluble fractions of Viola yedoensis[J].Chin Herb Med,2015,7(1):80-87.
[5]杜冬生,秦艳,程志红,等.紫花地丁的化学成分研究[J].中草药,2018,49(9):2007-2013.Du D S,Qin Y,Cheng Z H,et al.Chemical constituents of viola yedoensis[J].Chinese Traditional and Herbal Drugs,2018,49(9):2007-2013.
[6]曹捷.紫花地丁黄酮苷类成分的定性定量研究[D].上海:复旦大学,2013.
[7]Qin B,Chen Q,Shi LW,et al.Separation and quantitative determination of three coumarins in the chinese traditional drug zihua diding,herba violae,by high performance liquid chromatography[J].Journal of Chinese pharmaceutical Sciences,1994,3(2):157-163.
[8]中国科学院上海药物研究所.中草药有效成分提取与分离[M]·2版.上海:上海科学技术出版社,1983:156-177.
[9]李艳丽,胡彦武.紫花地丁抗炎作用及机制研究[J].中国实验方剂学杂志,2012,18(24):244-248.Li Y L,Hu Y W.Studies on anti-inflammatory effects and mechanism of viola yedoensis[J].Chinese Journal of Expe-]rimental Traditional Medical Formulae,2012,18(24):244-248.
[10]Xie C,Veitch N C,Houghton P J.Flavone C-glycosides from viola yedoensis makino[J].Chem Pharm Bull,2003,51(10):1204-1207.
[11]陈胡兰,董小萍,张梅,等.紫花地丁化学成分研究[J].中草药,2010,41(6):874-876.Chen H L,Dong X P,Zhang M,et al.Chemical constituents of viola yedoensis[J].Chinese Traditional and Herbal Drugs,2010,41(6):874-876.
[12]杨鹏鹏,闫福林,梁一兵,等.紫花地丁化学成分的研究[J].新乡医学院学报,2008,2(25):185-187.Yang P P,Yan Fulin,Liang Y B,et al.Study of Chemical constituents of Viola yedoensis[J].Journal of Xinxiang Medical College,2008,2(25):185-187.
[13]Wang C K,Colgrave M L,Gustafson K R,et al.Anti-HI-V cyclotides from the Chinese medicinal herb Viola yedoensis[J].J Nat Prod,2008,71:47-52.
[14]Xu J,Zhao Y,Aisa H A.Anti-inflammatory effect of pomegranate flower in lipopolysaccharide(LPS)-stimulated RAW264.7 macrophages[J].Pharm Biol,2017,55(1):2095-2101.
[15]Lee C W,Park S M,Zhao R,et al.Hederagenin,a major component of clematis mandshurica ruprecht root,attenuates inflammatory responses in RAW264.7 cells and in mice[J].Int Immuno pharmacol,2015,29(2):528-537.
[16]Plastina P,Benincasa C,Perri E,et al.Identification of hydroxytyrosyl oleate,a dsrivative of hydroxytyrosol with antiinflammatory properties,in olive oil by-products[J].Food Chem,2019,279:105-113.
[17]Park H J,Kim M M.Amentoflavone induces autophagy and modulates p53[J].Cell J,2019,21(1):27-34.
[18]Huang S F,Chu S C,Hsieh Y H,et al.Violay Yedoensis suppresses cell invasion by targeting the protease and NFκB activities in A549 and lewis lung carcinoma cells[J].Int J Med Sci,2018,15(4):280-290.
[19]Fernando I P S,Sanjeewa K K A,Samarakoon K W,et al.A fucoidan fraction purified from chnoospora minima,apotential inhibitor of LPS-induced inflammatory response[J].International Journal of Biological Macromolecules,2017,104(8):1185-1193.
[20]Jeong Y H,Oh Y C,Cho W K,et al.Anti-inflammatory effects of Viola yedoensis and the application of cell extraction methods for investigating bioactive constituents in macrophages[J].BMC Complement Altern Med,2016,16:180.