白花丹参内生真菌HBG16的抗凝血活性研究
|
摘要
目的:考察白花丹参内生真菌菌株HBG16的体外抗凝血活性。方法:采用形态学和分子方法进行属种鉴定;通过PDB培养基对HBG16进行液体发酵,采用甲醇、乙醇、乙酸乙酯和丙酮对菌丝体代谢产物进行提取,考察体外条件下甲醇、乙醇、乙酸乙酯和丙酮四种溶剂粗提物对家兔体外凝血时间(CT)、凝血酶时间(TT)、凝血酶原时间(PT)以及活化部分凝血酶时间(APTT)的影响。结果:白花丹参内生真菌HBG16经鉴定为尖孢镰刀菌;HBG16的菌丝甲醇粗提物对于内源性、外源性凝血都具有一定的延长作用,并且内源性凝血途径最为明显;丙酮、无水乙醇粗提物只作用于内源性凝血途径。结论:白花丹参内生真菌HBG16具有很好的抗凝血作用,可为研制安全无毒的新型抗凝血类药物提供新的资源和方法。
Objective:To investigate the anti-coagulation activity of endophytic fungus HBG16 isolated from Salvia miltiorrhiza Bge.f. alba in vitro. Methods:The endophytic fungus HBG16 was identified by the morphological and molecular identification methods and was fermented in PDB culture medium. Four extracts by different solvents(methanol,ethanol,ethyl acetate and acetone)were used to crude extract the mycelium metabolites,and the effects of crude extracts on coagulation time(CT),thrombin time(TT),prothrombin time(PT)and activation of partial thrombin time(APTT)were investigated. Results:Strain HBG16 was identified as Fusarium oxysporum. The methanol crude extract of HBG16 mycelia had extended effect on intrinsic and exogenous coagulation especially on intrinsic coagulation,while acetone and ethanol crude extract only act on intrinsic coagulation. Conclusion:Endophytic fungus HBG16 isolated from Salvia miltiorrhiza Bge.f.alba had significant anti-coagulation activity. It would provid new resources and methods for safe and nontoxic new anticoagulant drugs.
Objective:To investigate the anti-coagulation activity of endophytic fungus HBG16 isolated from Salvia miltiorrhiza Bge.f. alba in vitro. Methods:The endophytic fungus HBG16 was identified by the morphological and molecular identification methods and was fermented in PDB culture medium. Four extracts by different solvents(methanol,ethanol,ethyl acetate and acetone)were used to crude extract the mycelium metabolites,and the effects of crude extracts on coagulation time(CT),thrombin time(TT),prothrombin time(PT)and activation of partial thrombin time(APTT)were investigated. Results:Strain HBG16 was identified as Fusarium oxysporum. The methanol crude extract of HBG16 mycelia had extended effect on intrinsic and exogenous coagulation especially on intrinsic coagulation,while acetone and ethanol crude extract only act on intrinsic coagulation. Conclusion:Endophytic fungus HBG16 isolated from Salvia miltiorrhiza Bge.f.alba had significant anti-coagulation activity. It would provid new resources and methods for safe and nontoxic new anticoagulant drugs.
引文
[1]Taggart M. Vascular function in health anddisease review series[J].Journal of Cellular and Molecular Medicine,2010,14:1017.
[2]唐利,向毅,杜潇. 抗血栓药物临床应用研究进展[J].心血管病学进展,2016,37(6):672-677.
[3]胡大一,孙艺红. 血栓栓塞性疾病治疗新指南和新证据[J].中国实用内科杂志,2006,26(16):1293-1294.
[4]程忠玲,邵建明. 丝素蛋白作为抗凝血材料的研究与进展[J].中国组织工程研究与临床康复,2007,11(18):3621-3624.
[5]石凉,吴海洋. 蚕蛹壳聚糖甘氨酸氧化物的制备及其抗凝血作用研究[J].蚕业科学,2010,36(4):718-722.
[6]Wu W,Wang Y. Pharmacological actions and therapeutic application of Salvia miltiorrhiza depside salt and its active conponents[J]. Acta Pharmacologica Sinica,2012,33(9):1119-1130.
[7]王冰瑶,吴晓燕,樊官伟. 丹参素保护心血管系统的药理作用机制研究进展[J]. 中草药,2014,45(17):2571-2575.
[8]庞蕾,严铸云,梁俊玉,等.内生真菌在中药中的应用前景[J]. 时珍国医国药,2005,16(3):242-243.
[9]程玉鹏,李天聪,林进华. 产与宿主相同活性成分的内生真菌的研究进展[J]. 药物生物技术,2016,23(4):363-366.
[10]周茂金,苏美英,刘昌孝. 丹参素研究进展[J]. 中草药,2003,34(10):24-25.
[11]袁恒杰. 丹参素药理作用研究新进展[J]. 中国医学院药学杂志,2006,26(5):604-606.
[12]马丙祥,董宠凯. 丹参的药理作用研究新进展[J]. 中国药房,2014,25(7):663-665.
[13]刘艾林,李明源,王一涛,等. 丹参药理学活性物质基础研究现状[J]. 中国药学杂志,2007,42(9):641-646.
[14]李艳玲,史仁玖,郝岗平,等. 泰山产丹参内生真菌的分离鉴定和多样性分析[J]. 时珍国医国药,2012,23(1):114-117.
[15]王玉婷,李存能,石鹏亮,等. 丹参内生镰刀菌HBG16 胞外多糖发酵条件优化及其抗氧化能力分析[J]. 食品工业科技,2018(5):124-130.
[16]Li Y L,Xin X M,Chang Z Y,et al. The endophytic fungi of Salvia miltiorrhiza Bge.f alba are a potential source of natural antioxidants[J]. Botanical Studies,2015,56:1-7.
[17]魏景超. 真菌鉴定手册[M].北京:北京科学出版社,1979:196-197.
[18]吴海歌,姚子昂. 卡拉胶寡糖的抗凝血作用[J]. 中国食品添加剂试验究,2014,5:84-88.
[19]李德海,史锦硕,周聪,等. 黑木耳多糖的制备及其抗凝血功能的研究[J].安徽农业科学,2015,43(2):283-285.
[20]刘湘莲,张玉祥. 凝血PT APTT FIB TT的检验意义及其测定的有关事宜[J].内蒙古医学杂志,2004,36(5):369.
[21]Alban S,Schawerte A,Franz G. Anticoagulant sulfated polysaccha-rids:PartⅠ. Synthesis and structure-activity relationships of new pullulan sulfates[J].Carbohydrate Polymers,2002,47(2):267-276.
[2]唐利,向毅,杜潇. 抗血栓药物临床应用研究进展[J].心血管病学进展,2016,37(6):672-677.
[3]胡大一,孙艺红. 血栓栓塞性疾病治疗新指南和新证据[J].中国实用内科杂志,2006,26(16):1293-1294.
[4]程忠玲,邵建明. 丝素蛋白作为抗凝血材料的研究与进展[J].中国组织工程研究与临床康复,2007,11(18):3621-3624.
[5]石凉,吴海洋. 蚕蛹壳聚糖甘氨酸氧化物的制备及其抗凝血作用研究[J].蚕业科学,2010,36(4):718-722.
[6]Wu W,Wang Y. Pharmacological actions and therapeutic application of Salvia miltiorrhiza depside salt and its active conponents[J]. Acta Pharmacologica Sinica,2012,33(9):1119-1130.
[7]王冰瑶,吴晓燕,樊官伟. 丹参素保护心血管系统的药理作用机制研究进展[J]. 中草药,2014,45(17):2571-2575.
[8]庞蕾,严铸云,梁俊玉,等.内生真菌在中药中的应用前景[J]. 时珍国医国药,2005,16(3):242-243.
[9]程玉鹏,李天聪,林进华. 产与宿主相同活性成分的内生真菌的研究进展[J]. 药物生物技术,2016,23(4):363-366.
[10]周茂金,苏美英,刘昌孝. 丹参素研究进展[J]. 中草药,2003,34(10):24-25.
[11]袁恒杰. 丹参素药理作用研究新进展[J]. 中国医学院药学杂志,2006,26(5):604-606.
[12]马丙祥,董宠凯. 丹参的药理作用研究新进展[J]. 中国药房,2014,25(7):663-665.
[13]刘艾林,李明源,王一涛,等. 丹参药理学活性物质基础研究现状[J]. 中国药学杂志,2007,42(9):641-646.
[14]李艳玲,史仁玖,郝岗平,等. 泰山产丹参内生真菌的分离鉴定和多样性分析[J]. 时珍国医国药,2012,23(1):114-117.
[15]王玉婷,李存能,石鹏亮,等. 丹参内生镰刀菌HBG16 胞外多糖发酵条件优化及其抗氧化能力分析[J]. 食品工业科技,2018(5):124-130.
[16]Li Y L,Xin X M,Chang Z Y,et al. The endophytic fungi of Salvia miltiorrhiza Bge.f alba are a potential source of natural antioxidants[J]. Botanical Studies,2015,56:1-7.
[17]魏景超. 真菌鉴定手册[M].北京:北京科学出版社,1979:196-197.
[18]吴海歌,姚子昂. 卡拉胶寡糖的抗凝血作用[J]. 中国食品添加剂试验究,2014,5:84-88.
[19]李德海,史锦硕,周聪,等. 黑木耳多糖的制备及其抗凝血功能的研究[J].安徽农业科学,2015,43(2):283-285.
[20]刘湘莲,张玉祥. 凝血PT APTT FIB TT的检验意义及其测定的有关事宜[J].内蒙古医学杂志,2004,36(5):369.
[21]Alban S,Schawerte A,Franz G. Anticoagulant sulfated polysaccha-rids:PartⅠ. Synthesis and structure-activity relationships of new pullulan sulfates[J].Carbohydrate Polymers,2002,47(2):267-276.