黄芪和当归的主要活性成分配伍对骨髓抑制小鼠造血功能的影响
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摘要
目的探讨黄芪和当归的主要活性成分配伍对骨髓抑制小鼠模型的促造血作用。方法对芪归1∶1配伍提取物测定5种主要活性成分阿魏酸、芒柄花素、黄芪甲苷、毛蕊异黄酮、毛蕊异黄酮苷含量,以确定配伍剂量。小鼠随机分为空白对照组、模型组和各给药组。空白对照组给予溶剂灌胃7 d;模型组同上灌胃,于d 3腹腔注射环磷酰胺,连续3 d;各药物组灌胃给药,同上造模。检测外周血象、血清造血生长因子(HGF)含量、造血祖细胞集落数。结果芪归配伍提取物和活性成分配伍均可明显增加外周血白细胞(WBC)、红细胞(RBC)、血小板数(Pt)、血红蛋白(HGB)含量和骨髓造血组织面积,明显增加血清粒-巨噬细胞集落刺激因子(GM-CSF)、促血小板生成素(TPO)、促红细胞生成素(EPO)含量和骨髓粒-单核细胞集落形成单位(CFU-GM)、巨核细胞集落形成单位(CFU-MK)、红系集落形成单位(CFU-E)和爆式红系集落形成单位(BFU-E)数,两者作用相近。各成分中,阿魏酸可促进WBC和骨髓造血组织面积的恢复;5种成分均可促进RBC和HGB的恢复;阿魏酸和黄芪甲苷可促进Pt的恢复。5个成分均可升高TPO含量;阿魏酸和毛蕊异黄酮可升高GM-CSF含量;阿魏酸、毛蕊异黄酮和毛蕊异黄酮苷可升高EPO含量。除黄芪甲苷外,其它4种成分均可使CFU-GM增加;5种活性成分均可增加CFU-MK、CFU-E;仅阿魏酸可增加BFU-E。结论黄芪当归配伍发挥补血作用的主要药效物质是以上5种活性成分,这些活性成分配伍对促进造血可发挥增效作用。
Aim To explore the effect of the main active components combination of Astragalus promoting hematopoiesis on mouse model of bone marrow hematopoietic suppression. Methods The contents of five main active components such as ferulic acid, astragaloside Ⅳ, formononetin, calycosin and calycosin glycoside were determined after the extract of Astragalus combined with Angelica at 1 ∶1 ratio was prepared by water extraction, thus ascertaining the combination dosage. Mice were randomly divided into blank control, model, active component alone, five active components? combination and Astragalus-Angelica combination groups. The blank control group was given solvent for 7 days; the model group was given the intragastric administration as above and received intraperitoneal injection of cyclophosphamide on day 3 for 3 days; each drug group was treated intragastrically and established the model as above. The peripheral blood, the content of serum hematopoietic growth factor and the colony forming of hematopoietic progenitor cells were measured. Results The active components? combination and Astragalus-Angelica combination could significantly increase the numbers of peripheral blood leukocytes, erythrocytes, platelets, the content of hemoglobin and the area of bone marrow hematopoietic tissue(P<0.05 or 0.01); five active components could promote the recovery of erythrocytes and hemoglobin; ferulic acid could promote the recovery of leukocyte number and bone marrow hematopoietic tissue area; ferulic acid and astragaloside IV could promote the recovery of platelet. The contents of granulocyte-macrophage colony-stimulating factor(GM-CSF), thrombopoietin(TPO), erythropoietin(EPO) in serum significantly increased in Astragalus-Angelica combination and the active components combination groups(P<0.05 or 0.01); five components could increase TPO content, ferulic acid and calycosin could increase the contents of GM-CSF and EPO, calycosin glycoside could increase EPO.Astragalus-Angelica combination and the active components combination could significantly increase the colony numbers of granulocyte-monocyte colony foming unit(CFU-GM),megakaryocyte colony forming unit(CFU-MK), erythroid colony forming unit(CFU-E), burst forming unit-erythroid(BFU-E);except for astragaloside Ⅳ,the others could increase the number of CFU-GM, five active components could all increase the numbers of CFU-MK,CFU-E,but only ferulic acid could increase the number of BFU-E.Conclusions The main active substances that the combination of Astragalus and Angelica enriches the blood are the above five active components, which exert the synergistic effect through acting on different links of hematopoiesis such as increasing the content of hematopoietic growth factor in blood, promoting the proliferation of hematopoietic progenitor cells,and so on.
Aim To explore the effect of the main active components combination of Astragalus promoting hematopoiesis on mouse model of bone marrow hematopoietic suppression. Methods The contents of five main active components such as ferulic acid, astragaloside Ⅳ, formononetin, calycosin and calycosin glycoside were determined after the extract of Astragalus combined with Angelica at 1 ∶1 ratio was prepared by water extraction, thus ascertaining the combination dosage. Mice were randomly divided into blank control, model, active component alone, five active components? combination and Astragalus-Angelica combination groups. The blank control group was given solvent for 7 days; the model group was given the intragastric administration as above and received intraperitoneal injection of cyclophosphamide on day 3 for 3 days; each drug group was treated intragastrically and established the model as above. The peripheral blood, the content of serum hematopoietic growth factor and the colony forming of hematopoietic progenitor cells were measured. Results The active components? combination and Astragalus-Angelica combination could significantly increase the numbers of peripheral blood leukocytes, erythrocytes, platelets, the content of hemoglobin and the area of bone marrow hematopoietic tissue(P<0.05 or 0.01); five active components could promote the recovery of erythrocytes and hemoglobin; ferulic acid could promote the recovery of leukocyte number and bone marrow hematopoietic tissue area; ferulic acid and astragaloside IV could promote the recovery of platelet. The contents of granulocyte-macrophage colony-stimulating factor(GM-CSF), thrombopoietin(TPO), erythropoietin(EPO) in serum significantly increased in Astragalus-Angelica combination and the active components combination groups(P<0.05 or 0.01); five components could increase TPO content, ferulic acid and calycosin could increase the contents of GM-CSF and EPO, calycosin glycoside could increase EPO.Astragalus-Angelica combination and the active components combination could significantly increase the colony numbers of granulocyte-monocyte colony foming unit(CFU-GM),megakaryocyte colony forming unit(CFU-MK), erythroid colony forming unit(CFU-E), burst forming unit-erythroid(BFU-E);except for astragaloside Ⅳ,the others could increase the number of CFU-GM, five active components could all increase the numbers of CFU-MK,CFU-E,but only ferulic acid could increase the number of BFU-E.Conclusions The main active substances that the combination of Astragalus and Angelica enriches the blood are the above five active components, which exert the synergistic effect through acting on different links of hematopoiesis such as increasing the content of hematopoietic growth factor in blood, promoting the proliferation of hematopoietic progenitor cells,and so on.
引文
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[13] Gong A G,Lau K M,Xu M L,et al.The estrogenic properties of Danggui Buxue Tang,a Chinese herbal decoction,are triggered predominantly by calycosin in MCF-7 cells[J].J Ethnopharmacol,2016,189:81-9.
[14] Yang M,Chan G C,Deng R,et al.An herbal decoction of Radix astragali and Radix angelicae sinensis promotes hematopoiesis and thrombopoiesis[J].J Ethnopharmacol,2009,124(1):87-97.
[15] 尹利明,赵燕娜,杜文喜,等.人参总皂苷增强成骨分化的间充质干细胞促造血作用研究[J].中国药理学通报,2015,31(1):45-9.[15] Yin L M,Zhao Y N,Du W X,et al.Total saponins of panax ginseng enhances the effect of osteoblast differentiation from mesenchymal stem cell on promoting hematopoiesis[J].Chin Pharmacol Bull,2015,31(1):45-9.
[16] Gallicchio V S,Gasale G P,Watts T.Inhibition of human bone marrow-derived stem cell colony formation (CFU-E,BFU-E,and CFU-GM) following in vitro exposure to organophosphates[J].Exp Hematol,1987,15(11):1099-102.
[2] Yan S,Xie Y,Zhu B,et al.Effect comparison of different formulation of Dang-Gui-Bu-Xue-Tang on myelosuppression mouse[J].Asian Pac J Trop Med,2011,4(7):556-9.
[3] 史旭芹,尚尔鑫,唐于平,等.基于响应曲面分析法对当归-黄芪配伍养血补血功效相互作用研究[J].药学学报,2012,47(10):1375-83.[3] Shi X Q,Shang E X,Tang Y P,et al.Interaction of nourishing and tonifying blood effects of the combination of Angelicae sinensis Radix and Astragali Radix studied by response surface method[J].Acta Pharm Sin,2012,47(10):1375-83.
[4] Li F,Tang R,Chen L B,et al.Effects of Astragalus combined with Angelica on bone marrow hematopoiesis suppression induced by cyclophosphamide in mice[J].Biol Pharm Bull,2017,40(5):598-609.
[5] 唐蓉,张依人,陈叶童,等.不同剂量配伍对黄芪-当归中5种活性成分的影响[J].中国实验方剂学杂志,2016,22(23):1-5.[5] Tang R,Zhang Y R,Chen Y T,et al.Effect of different compatibility on five chemical components in Astragali radix-Angelicae sinensis radix[J].Chin J Exp Tradit Med Formul,2016,22(23):1-5.
[6] 齐丽娟,宋雁,王伟,等.用环磷酰胺建立小鼠免疫抑制动物模型[J].卫生研究,2010,39(3):313-5.[6] Qi L J,Song Y,Wang W,et al.Comparison of immunosuppression induced by different doses of cyclophosphamide in normal mice[J].J Hyg Res,2010,39(3):313-5.
[7] Shi X,Tang Y,Zhu H,et al.Comparative tissue distribution profiles of five major bioactive components in normal and blood deficiency rats after oral administration of Danggui Buxue decoction by UPLC-TQ/MS[J].J Pharm Biomed Anal,2014,88:207-15.
[8] Tabbara I A.Erythropoietin.Biology and clinical applications[J].Arch Inter Med,1993,153(3):298-304.
[9] Metcalf D.Hematopoietic regulators:redundancy or subtlety[J]?Blood,1993,82(12):3515-23.
[10] Kaushansky K.Thrombopoiesis[J].Semin Hematol,2015,52(1):4-11.
[11] Gao Q T,Cheung J K,Choi R C,et al.A Chinese herbal decoction prepared from Radix Astragali and Radix Angelicae Sinensis induces the expression of erythropoietin in cultured Hep3B cells[J].Planta Med,2008 ,74(4):392-5.
[12] Zheng K Y,Choi R C,Xie H Q,et al.The expression of erythropoietin triggered by Danggui Buxue Tang,a Chinese herbal decoction prepared from radix Astragali and radix Angelicae Sinensis,is mediated by the hypoxia-inducible factor in cultured HEK293T cells[J].J Ethnopharmacol,2010,132(1):259-67.
[13] Gong A G,Lau K M,Xu M L,et al.The estrogenic properties of Danggui Buxue Tang,a Chinese herbal decoction,are triggered predominantly by calycosin in MCF-7 cells[J].J Ethnopharmacol,2016,189:81-9.
[14] Yang M,Chan G C,Deng R,et al.An herbal decoction of Radix astragali and Radix angelicae sinensis promotes hematopoiesis and thrombopoiesis[J].J Ethnopharmacol,2009,124(1):87-97.
[15] 尹利明,赵燕娜,杜文喜,等.人参总皂苷增强成骨分化的间充质干细胞促造血作用研究[J].中国药理学通报,2015,31(1):45-9.[15] Yin L M,Zhao Y N,Du W X,et al.Total saponins of panax ginseng enhances the effect of osteoblast differentiation from mesenchymal stem cell on promoting hematopoiesis[J].Chin Pharmacol Bull,2015,31(1):45-9.
[16] Gallicchio V S,Gasale G P,Watts T.Inhibition of human bone marrow-derived stem cell colony formation (CFU-E,BFU-E,and CFU-GM) following in vitro exposure to organophosphates[J].Exp Hematol,1987,15(11):1099-102.