大接骨丹免疫调节活性成分研究
|
摘要
湖北民间草药大接骨丹为山茱萸科鞘柄木属植物有齿鞘柄木(Toricellia an-gulata Oliv. var. intermedia (Harms) Hu)的根、根皮、茎皮、叶及花,可治疗外伤骨折、跌打损伤、风湿关节痛、干血痨伤、产后腰痛、慢性肠炎、腹泻、扁桃腺炎等。该药在湖北分布较广,恩施民间多用其治疗风湿关节痛,疗效确切,具有广阔的开发前景。
本实验对大接骨丹95%乙醇提取物的石油醚部位和正丁醇部位进行色谱分离,得到13个化合物。通过理化常数、各种波谱分析技术或直接与标准品对照,鉴定其中的12个化合物分别为:二(4,7,8-三羟基-3,5-二甲氧基苯丙基)-9-醚(1,新化合物)、豆甾醇(2)、a-3,4,5-四羟基-苯乙酸(3)、豆甾-5,11-二烯-3β-醇(4)、22,23-二氢豆甾醇(5)、β-胡萝卜苷(6)、26,27-dinorcholesta-8,14-dien-3β-ol(7)、十六烷酸(8)、十八烷酸(9)、α-D-吡喃葡萄糖(10)、β-D-吡喃葡萄糖(11)、4-羟基-3,5-二甲氧基苯甲醛(12)。其中化合物1为新化合物;化合物3为首次从天然植物中提取分离得到;化合物1、2、3、7、12均为首次从该植物中分离得到。
对大接骨丹中分离得到的六个单体化合物1、2、5、6、7、13进行了与RA发生相关的免疫活性实验—Con A诱导大鼠脾细胞增殖反应实验。结果表明,与对照组比较由大接骨丹分离所得这六个单体化合物对Con A诱导大鼠脾细胞增殖均有不同程度的抑制作用,差异具有统计学意义(P < 0.05)。
Toricellia angulata Oliv. var. intermedia (Harms) Hu is the root or flower of Toothedleaf Torricellia of Cornaceae, used as a folk medicine in Hubei province for surgical trauma,fracture,rheumatic arthritis,blood stasis and tabes, puerperal lumbago, chronic enteritis, diarrhoea and tonsillitis. Toricellia angulata Oliv. with its excellent effect is widely used to treat rheumatic arthritis in Enshi of Hubei province, and it have an extensive perspective of development.
During our investigation of the chemical constituents of this plant, thirteencompounds have been isolated from the petroleum ether, n-BuOH-soluble porti-ons in the 95% ethanol extract. On the basis of spectral analysis or direct co- chromatographies with authentic samples, twleve of them were identified as Di-(4,7,8-trihydroxy-3,5-dimethoxyl phenylpropyl )-9-ether (1, new compound), Stigmasterol (2), a-3,4,5-tetrahydroxy benzeneacetic acid (3), Stigmasta-5,11- dien-3β-ol (4), 22,23-dihydrostigmasterol (5),β-daucosterol (6),26,27-dinorchol-esta-8,14-dien-3β-ol (7), Palmitic acid (8), Octadecanoic acid(9),α-D-glucopy-ranose (10),β-D-glucopyranose (11), 4-hydroxy-3,5-dimethoxyl benzaldehyde (12). The compound 1 was a new compound, compound 3 was isolated from nature plant for the first time and compounds 1, 2, 3, 7, 12 were isolated from thisplant for the first time.
Compounds 1, 2, 5, 6, 7, 13 were studied var experiments of immunological activity related to the creation of rheumatic arthritis—Con A-induced cell proliferation of rats splenocytes. Compared with the control group, those compounds have immunological inhibiting effect obviously on Con A-induced cell proliferation of rats splenocytes(P < 0.05).
本实验对大接骨丹95%乙醇提取物的石油醚部位和正丁醇部位进行色谱分离,得到13个化合物。通过理化常数、各种波谱分析技术或直接与标准品对照,鉴定其中的12个化合物分别为:二(4,7,8-三羟基-3,5-二甲氧基苯丙基)-9-醚(1,新化合物)、豆甾醇(2)、a-3,4,5-四羟基-苯乙酸(3)、豆甾-5,11-二烯-3β-醇(4)、22,23-二氢豆甾醇(5)、β-胡萝卜苷(6)、26,27-dinorcholesta-8,14-dien-3β-ol(7)、十六烷酸(8)、十八烷酸(9)、α-D-吡喃葡萄糖(10)、β-D-吡喃葡萄糖(11)、4-羟基-3,5-二甲氧基苯甲醛(12)。其中化合物1为新化合物;化合物3为首次从天然植物中提取分离得到;化合物1、2、3、7、12均为首次从该植物中分离得到。
对大接骨丹中分离得到的六个单体化合物1、2、5、6、7、13进行了与RA发生相关的免疫活性实验—Con A诱导大鼠脾细胞增殖反应实验。结果表明,与对照组比较由大接骨丹分离所得这六个单体化合物对Con A诱导大鼠脾细胞增殖均有不同程度的抑制作用,差异具有统计学意义(P < 0.05)。
Toricellia angulata Oliv. var. intermedia (Harms) Hu is the root or flower of Toothedleaf Torricellia of Cornaceae, used as a folk medicine in Hubei province for surgical trauma,fracture,rheumatic arthritis,blood stasis and tabes, puerperal lumbago, chronic enteritis, diarrhoea and tonsillitis. Toricellia angulata Oliv. with its excellent effect is widely used to treat rheumatic arthritis in Enshi of Hubei province, and it have an extensive perspective of development.
During our investigation of the chemical constituents of this plant, thirteencompounds have been isolated from the petroleum ether, n-BuOH-soluble porti-ons in the 95% ethanol extract. On the basis of spectral analysis or direct co- chromatographies with authentic samples, twleve of them were identified as Di-(4,7,8-trihydroxy-3,5-dimethoxyl phenylpropyl )-9-ether (1, new compound), Stigmasterol (2), a-3,4,5-tetrahydroxy benzeneacetic acid (3), Stigmasta-5,11- dien-3β-ol (4), 22,23-dihydrostigmasterol (5),β-daucosterol (6),26,27-dinorchol-esta-8,14-dien-3β-ol (7), Palmitic acid (8), Octadecanoic acid(9),α-D-glucopy-ranose (10),β-D-glucopyranose (11), 4-hydroxy-3,5-dimethoxyl benzaldehyde (12). The compound 1 was a new compound, compound 3 was isolated from nature plant for the first time and compounds 1, 2, 3, 7, 12 were isolated from thisplant for the first time.
Compounds 1, 2, 5, 6, 7, 13 were studied var experiments of immunological activity related to the creation of rheumatic arthritis—Con A-induced cell proliferation of rats splenocytes. Compared with the control group, those compounds have immunological inhibiting effect obviously on Con A-induced cell proliferation of rats splenocytes(P < 0.05).
引文
[1]叶任高,陆再英.内科学[M].人民卫生出版社,第五版,879-903.
[2] G. Zhang. The progression of research on cell apoptosis molecule of rheumatoid arthritis[J]. International Journal of Immunology,2006,29(3).
[3]American college of rheumatology subcommittee on eumatoid arthritis guidelines.Guidelines for the management of rheumatoid arthritis[M],2002 update. Arthritis Rheum,2002,46(2):328-346.
[4]A1etaha D., Smolen J. S., Effectiveness profiles and dose dependent retention of traditional disease modifying antirheumatic drugs for rheumatoid arthritis[J].An observational study.J Rheumato1,2002(29):1631-1638.
[5] C. Q. Zhong, G. X. CAO. The immunosuppressant of traditional Chinese medicine[J]. Qilu Pharmaceutical Affairs,2007,126(16):359-360.
[6]尹新芹,张东升.雷公藤临床作用进展[J].医学综述,2001,7(7):438～4391.
[7]杨济秋,杨济中.贵州民间方药集[M].贵阳:贵州人民出版社,1961(56).
[8]胡文光.中国植物志(第56卷)[M].北京:科学出版社,1990.
[9] S. Wu, Y. Ma, et al. The chemical constituents from Toricellia angulata[J]. Acta Botanica Yunnanica,2000,22(2):214-218.
[10]王晶.大接骨丹的免疫活性成分研究.华中科技大学硕士毕业论文,2008.5.
[11]贾于.大接骨丹抗炎镇痛活性成分研究.华中科技大学硕士毕业论文,2008.5.
[12]Hideyuki Matsuura,Hiroyuki Miyazaki,Chikako Asakawa,et al. Isolation ofα-glusosidase inhibitors from hyssop[J]. Phytochemistry, 2004(65):91-97.
[13]Hisashi K, Noriko S, Akiko H, et al. Sterol glucosides from prunella vulgaris[J].Phytochemistry,1990,29(7):2351-2355.
[14] Goodwin, Brian L, Ruthven, Colin R. J. et al. Metabolism of phenylethanolamines and 2-oxo-2-phenylethylamines in the rat[J]. General Pharmacology, 1997,28(4):535-543.
[15]李君竹,刘海洋等.角果胡椒的化学成分研究[J].天然产物研究与开发,2007(19):620-622,630.
[16]Institute of Medical Plant Development Chinese Academy of Medical Scie-nce and Peking Union Medical College Record of Chinese Materia Medical[M]. Beijing: People’s Medical publishing House,1960:290-292.
[17]崔红花,高幼衡.川佛手化学成分研究(Ⅰ).中草药,2007,38(9).
[18] X. W. Dong, L. C. Zhong. A Dimeric Sinapaldehyde Glucoside from Ilex Rotunda[J]. Phytochemistry,1996,41(2):657-659.
[19]黄岚,冯树芳等.雷公藤内酯醇对体外培养的成骨细胞增殖的影响[J].中华皮肤科杂志,2000(33).
[20]H. Gerhard Vogel, Wolfgang H. Vogel. Drug Discovery Evaluation-Phamaco- logical Assays[M]. 1997
[21]徐叔云,卞如濂等.药理实验方法学[M].第三版.人民卫生出版社,2002, 1421-1422.
[22] F. H. Zhang, S. J. Li, et al. Effect of CCK-8 on B7-1 and B7-2 expressions and costimulatory activity of LPS-activated murine macrophages[J]. Chinese Pharmacological Bulletin,2007,23 (10):1271-1275.
[23]H. Y. Hu, Z. Y. Song et al. Regulation of 3'-meisoindigo on proliferation,apoptosis and biological function of splenocytes from diferent germline mice[J]. Journal of Clinical Rehabilitative Tissue Engineering Research ,2008,12(34):6709-6713.
[1]胡文光.中国植物志(第56卷)[M].北京:科学出版社,1990.
[2]何雨祥.珍稀木本油料植物—光皮树[J].福建农业科技,1991(1).
[3]候家麟.三十三种常用补益类中药材微量元素测定[J].甘肃药学,l991(1):1.
[4]陈训,巫华美.川鄂山茱萸染色体研究[J].广西植物,1992(4).
[5]潘扬,王天山.植物山茱萸化学成分的研究概况[J].南京中医药大学学报, 1998(14):61-62.
[6]祝美云,黄忠民等.山茱萸保健饮料的研制[J].郑州轻工业学院学报,1998,13(2):66-70.
[7]H. Zhu. A revision of the Genus Mastixia (Cornaceae) of China[J]. Acta Botanica Yunnanica,1993,15(2):13l-133.
[8]王洪.中国特有野生植物—长尾单室茱萸[J].中国野生植物资源,1986(6):21-22.
[9]H. Zhu. A New Plant Species from Vietnam[J]. Bulletin of Botanical Res-earch,2000,20(2):129-130.
[10]W. P. Fang. Comaceae[M].In:Flora Reipublicae Popularis Sinicae.Beijing:Science Press,1990,56:38-l08.
[11]李峰.特色观叶花卉品种及特性[J].云南科技报,2001,7(2).
[12]李泽相.怎样培植特色观叶花卉[J].四川农业科技,2002(5).
[13]钟园.认识抗污染植物[J].浙江林业—园林绿化,2005(2):29-31.
[14]S. M. Gao, P. J. Cao. Study on cold acclimation and freezing-tolerance mechanism of Aucuba japonica CV.Variegata[J]. Aeta Bat. Borea1. OccidentSm.,2003,23(12):211-213.
[15]W. L. Gao, S. M. Gao, The initial report of the study of the photosynth-esis and protein in Acuba jap variegte under low temperature[J]. Acta Agricu-hurae Universitatis Jiangxiensis,24(3):376-379.
[16]彭创业,林静等.桃叶珊瑚的生药学研究[J].时珍国医国药,2008,l9(l0).
[17]陈西仓,姚德生.奇异花木[J].特种经济动植物,2002(4).
[18]陈西仓,张振纲.四照花的利用和繁育技术[J].林业科技开发:2003,l7(3): 50-51.
[19]李昌珠,赵江红.生物油料能源树种—光皮树[J].湖南林业,2007(2):48.
[20]梁仰贞.光皮树栽培技术[J].特种经济动植物,2007(3):38-39.
[21]R. C. Hosie. Native trees of Canada[M]. Supply&Seniecs:Canada Publishi-ng Center,1979.
[22]王雅琴,毕艳春.长春市园林绿化的新品种—偃伏梾木[J]. 2007(5):145-146.
[23]杨娟,娄方明等.中华青荚叶化学成分研究[J].中国中药杂志,2007,32(14):1416-1418.
[24]G. F. Li, P. J. Zhu, et al. A New Fructofuranoside from Helwingia chi-nensis(Cornaceae)[J]. Aeta Botaniea Yunnaniea,2008,30(1):115-120.
[25]G. F. Li, Y. F. Wang, et al. A New Secoiridoid Glycoside from Helwingia chinensis(Cornaceae)[J]. Aeta Botaniea Yunnaniea,2006,28(6):676-678.
[26]汪建云,孔治有等.西域青荚叶实生苗驯化的初步研究[J].中国野生植物资源,2005,24(4):59-62.
[27]杨济秋,杨济中.贵州民间方药集[M].贵阳:贵州人民出版社,1961(56).
[28]石福臣,尹淑兰.小兴安岭的草茱萸[J].国土与自然资源研究,1991(1):56-57.
[29]Y. B. Chen. A little known plant of China[J]. Bull. Bot. Lab. North-East. Forest. Inst.,1979(4):47-49.
[2] G. Zhang. The progression of research on cell apoptosis molecule of rheumatoid arthritis[J]. International Journal of Immunology,2006,29(3).
[3]American college of rheumatology subcommittee on eumatoid arthritis guidelines.Guidelines for the management of rheumatoid arthritis[M],2002 update. Arthritis Rheum,2002,46(2):328-346.
[4]A1etaha D., Smolen J. S., Effectiveness profiles and dose dependent retention of traditional disease modifying antirheumatic drugs for rheumatoid arthritis[J].An observational study.J Rheumato1,2002(29):1631-1638.
[5] C. Q. Zhong, G. X. CAO. The immunosuppressant of traditional Chinese medicine[J]. Qilu Pharmaceutical Affairs,2007,126(16):359-360.
[6]尹新芹,张东升.雷公藤临床作用进展[J].医学综述,2001,7(7):438～4391.
[7]杨济秋,杨济中.贵州民间方药集[M].贵阳:贵州人民出版社,1961(56).
[8]胡文光.中国植物志(第56卷)[M].北京:科学出版社,1990.
[9] S. Wu, Y. Ma, et al. The chemical constituents from Toricellia angulata[J]. Acta Botanica Yunnanica,2000,22(2):214-218.
[10]王晶.大接骨丹的免疫活性成分研究.华中科技大学硕士毕业论文,2008.5.
[11]贾于.大接骨丹抗炎镇痛活性成分研究.华中科技大学硕士毕业论文,2008.5.
[12]Hideyuki Matsuura,Hiroyuki Miyazaki,Chikako Asakawa,et al. Isolation ofα-glusosidase inhibitors from hyssop[J]. Phytochemistry, 2004(65):91-97.
[13]Hisashi K, Noriko S, Akiko H, et al. Sterol glucosides from prunella vulgaris[J].Phytochemistry,1990,29(7):2351-2355.
[14] Goodwin, Brian L, Ruthven, Colin R. J. et al. Metabolism of phenylethanolamines and 2-oxo-2-phenylethylamines in the rat[J]. General Pharmacology, 1997,28(4):535-543.
[15]李君竹,刘海洋等.角果胡椒的化学成分研究[J].天然产物研究与开发,2007(19):620-622,630.
[16]Institute of Medical Plant Development Chinese Academy of Medical Scie-nce and Peking Union Medical College Record of Chinese Materia Medical[M]. Beijing: People’s Medical publishing House,1960:290-292.
[17]崔红花,高幼衡.川佛手化学成分研究(Ⅰ).中草药,2007,38(9).
[18] X. W. Dong, L. C. Zhong. A Dimeric Sinapaldehyde Glucoside from Ilex Rotunda[J]. Phytochemistry,1996,41(2):657-659.
[19]黄岚,冯树芳等.雷公藤内酯醇对体外培养的成骨细胞增殖的影响[J].中华皮肤科杂志,2000(33).
[20]H. Gerhard Vogel, Wolfgang H. Vogel. Drug Discovery Evaluation-Phamaco- logical Assays[M]. 1997
[21]徐叔云,卞如濂等.药理实验方法学[M].第三版.人民卫生出版社,2002, 1421-1422.
[22] F. H. Zhang, S. J. Li, et al. Effect of CCK-8 on B7-1 and B7-2 expressions and costimulatory activity of LPS-activated murine macrophages[J]. Chinese Pharmacological Bulletin,2007,23 (10):1271-1275.
[23]H. Y. Hu, Z. Y. Song et al. Regulation of 3'-meisoindigo on proliferation,apoptosis and biological function of splenocytes from diferent germline mice[J]. Journal of Clinical Rehabilitative Tissue Engineering Research ,2008,12(34):6709-6713.
[1]胡文光.中国植物志(第56卷)[M].北京:科学出版社,1990.
[2]何雨祥.珍稀木本油料植物—光皮树[J].福建农业科技,1991(1).
[3]候家麟.三十三种常用补益类中药材微量元素测定[J].甘肃药学,l991(1):1.
[4]陈训,巫华美.川鄂山茱萸染色体研究[J].广西植物,1992(4).
[5]潘扬,王天山.植物山茱萸化学成分的研究概况[J].南京中医药大学学报, 1998(14):61-62.
[6]祝美云,黄忠民等.山茱萸保健饮料的研制[J].郑州轻工业学院学报,1998,13(2):66-70.
[7]H. Zhu. A revision of the Genus Mastixia (Cornaceae) of China[J]. Acta Botanica Yunnanica,1993,15(2):13l-133.
[8]王洪.中国特有野生植物—长尾单室茱萸[J].中国野生植物资源,1986(6):21-22.
[9]H. Zhu. A New Plant Species from Vietnam[J]. Bulletin of Botanical Res-earch,2000,20(2):129-130.
[10]W. P. Fang. Comaceae[M].In:Flora Reipublicae Popularis Sinicae.Beijing:Science Press,1990,56:38-l08.
[11]李峰.特色观叶花卉品种及特性[J].云南科技报,2001,7(2).
[12]李泽相.怎样培植特色观叶花卉[J].四川农业科技,2002(5).
[13]钟园.认识抗污染植物[J].浙江林业—园林绿化,2005(2):29-31.
[14]S. M. Gao, P. J. Cao. Study on cold acclimation and freezing-tolerance mechanism of Aucuba japonica CV.Variegata[J]. Aeta Bat. Borea1. OccidentSm.,2003,23(12):211-213.
[15]W. L. Gao, S. M. Gao, The initial report of the study of the photosynth-esis and protein in Acuba jap variegte under low temperature[J]. Acta Agricu-hurae Universitatis Jiangxiensis,24(3):376-379.
[16]彭创业,林静等.桃叶珊瑚的生药学研究[J].时珍国医国药,2008,l9(l0).
[17]陈西仓,姚德生.奇异花木[J].特种经济动植物,2002(4).
[18]陈西仓,张振纲.四照花的利用和繁育技术[J].林业科技开发:2003,l7(3): 50-51.
[19]李昌珠,赵江红.生物油料能源树种—光皮树[J].湖南林业,2007(2):48.
[20]梁仰贞.光皮树栽培技术[J].特种经济动植物,2007(3):38-39.
[21]R. C. Hosie. Native trees of Canada[M]. Supply&Seniecs:Canada Publishi-ng Center,1979.
[22]王雅琴,毕艳春.长春市园林绿化的新品种—偃伏梾木[J]. 2007(5):145-146.
[23]杨娟,娄方明等.中华青荚叶化学成分研究[J].中国中药杂志,2007,32(14):1416-1418.
[24]G. F. Li, P. J. Zhu, et al. A New Fructofuranoside from Helwingia chi-nensis(Cornaceae)[J]. Aeta Botaniea Yunnaniea,2008,30(1):115-120.
[25]G. F. Li, Y. F. Wang, et al. A New Secoiridoid Glycoside from Helwingia chinensis(Cornaceae)[J]. Aeta Botaniea Yunnaniea,2006,28(6):676-678.
[26]汪建云,孔治有等.西域青荚叶实生苗驯化的初步研究[J].中国野生植物资源,2005,24(4):59-62.
[27]杨济秋,杨济中.贵州民间方药集[M].贵阳:贵州人民出版社,1961(56).
[28]石福臣,尹淑兰.小兴安岭的草茱萸[J].国土与自然资源研究,1991(1):56-57.
[29]Y. B. Chen. A little known plant of China[J]. Bull. Bot. Lab. North-East. Forest. Inst.,1979(4):47-49.