三七总皂苷对斑马鱼胚胎初级和次级造血的抑制作用
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摘要
目的探讨三七总皂苷(Panax notoginsenosides,PNSs)对斑马鱼胚胎造血的作用,为三七的药理学应用提供实验依据。方法通过乙醇提取得到PNSs,用50、100μg/mL的PNSs从75%外包时期开始处理斑马鱼胚胎。收集发育至不同时期的胚胎,检测药物处理后,斑马鱼初级造血和次级造血的分子标记的变化。结果 PNSs处理后,初级造血的分子标记gata1、hbbe3明显下降,生成的红细胞显著减少;PNSs处理还可以抑制造血干细胞(hematopoietic stem cell, HSC)发育。次级造血的分子标记runx1,cmyb在PNSs处理下表达下调,由HSC分化生成的T淋巴细胞分子标记rag1表达也显著降低。有意思的是,PNSs对斑马鱼初级和次级造血的抑制作用均呈剂量依赖效应。结论孕期可能需要慎重使用三七总皂苷药物。
Objective To investigate the effects of Panax notoginsenosides(PNSs) on primitive and definitive hematopoiesis of zebrafish embryos, and provide new insights into pharmacological application of PNSs. Methods PNSs were obtained by ethanol extraction from panax notoginseng powder. Zebrafish embryos were treated with 50 or 100 μg/mL PNSs from the 75% epiboly stage until harvest to detect hematopoietic markers by in situ hybridization. Results Compared with the control larvae, the embryos treated with PNSs exhibited obviously reduced expression of primitive hematopoietic markers such as gata1 and hbbe3. Consistently, the formation of erythrocytes was also impaired in the PNSs-treated embryos at later stages. Furthermore, PNSs treatments disrupted the definitive hematopoiesis as indicated by decreased expression of hematopoietic stem cell markers runx1 and cmyb and the T lymphocyte marker rag1. Interestingly, the inhibitory effects of PNSs on the primitive and definitive hematopoiesis were concentration-dependent. Conclusions Our results indicate that PNSs impair primitive and definitive hematopoiesis during zebrafish embryonic development, implying that PNSs should be used with caution during pregnancy.
Objective To investigate the effects of Panax notoginsenosides(PNSs) on primitive and definitive hematopoiesis of zebrafish embryos, and provide new insights into pharmacological application of PNSs. Methods PNSs were obtained by ethanol extraction from panax notoginseng powder. Zebrafish embryos were treated with 50 or 100 μg/mL PNSs from the 75% epiboly stage until harvest to detect hematopoietic markers by in situ hybridization. Results Compared with the control larvae, the embryos treated with PNSs exhibited obviously reduced expression of primitive hematopoietic markers such as gata1 and hbbe3. Consistently, the formation of erythrocytes was also impaired in the PNSs-treated embryos at later stages. Furthermore, PNSs treatments disrupted the definitive hematopoiesis as indicated by decreased expression of hematopoietic stem cell markers runx1 and cmyb and the T lymphocyte marker rag1. Interestingly, the inhibitory effects of PNSs on the primitive and definitive hematopoiesis were concentration-dependent. Conclusions Our results indicate that PNSs impair primitive and definitive hematopoiesis during zebrafish embryonic development, implying that PNSs should be used with caution during pregnancy.
引文
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[9] 吴韶,张瑜,杨春红,等.三七不同部位中止血活血成分三七皂苷 Fc 的HPLC测定 [J].中草药,2016,47(18):3289-3292.Wu S,Zhang Y,Yang CH,et al.HPLC determination of notoginsenoside Fc,a hemostatic and activating blood component,in different parts of Panax notoginseng [J].Chin Tradit Herbal Drug,2016,47(18):3289-3292.
[10] Ning G,Liu X,Dai M,et al.MicroRNA-92a upholds Bmp signaling by targeting noggin3 during pharyngeal cartilage formation [J].Dev Cell,2013,24(3):283-295.
[11] 李响.Mta3/NuRD复合体在斑马鱼初级造血过程中的功能研究 [D].清华大学,2010:56-57.Li X.Functional analysis of Mta3/NuRD complex in zebrafish primitive hematopoiesis [D].Tsinghua University,2010:56-57.
[12] 陈磊,刘怡.模式生物斑马鱼在中药研究中的应用 [J].药学学报,2012,47(4):434-439.Chen L,Liu Y.Application of zebrafish model organism in the research of Chinese materia medica [J].Acta Pharm Sin,2012,47(4):434-439.
[13] 高瑞兰,徐卫红,林筱洁,等.三七皂甙对造血细胞GATA-1 和GATA-2 转录调控蛋白的诱导作用 [J].中华血液学杂志,2004,25(5):281-284.Gao RL,Xu WH,Lin XJ,et al.Up-regulation of transcription factors GATA-1 and GATA-2 induced by Panax notoginosides in hematopoietic cells [J].Chin J Hematol,2004,25(5):281-284.
[2] Orkin SH,Zon LI.Hematopoiesis:an evolving paradigm for stem cell biology [J].Cell,2008,132(4):631-644.
[3] 王本祥.现代中药药理与临床 [M].天津:天津科技翻译出版公司,2004:79Wang BX.Modern Pharmacology and Clinic of Chinese Traditional Medicine [M].Tianjin:Tianjin Science & Technology Translation & Publishing Company,2004:79.
[4] 古丽丽,王慧敏.三七总皂苷对血液及造血系统的药理作用研究进展 [J].首都医药,2012 (10):37-38.Gu LL,Wang HM.Research progress on pharmacological action of blood and hematopoietic system induced by Panax notoginosides [J].Cap Med,2012 (10):37-38.
[5] 郭爽,张强,周伟龙,等.三七及其制剂联合化学药物防治心血管疾病的研究进展 [J].中华中医药杂志,2018,33(12):5518-5522.Guo S,Zhang Q,Zhou WL,et al.Research progress of Radix Notoginseng and its preparation combined with chemical drugs in prevention and treatment of cardiovascular diseases [J].Chin J Tradit Chin Med Pharm,2018,33(12):5518-5522.
[6] 刘咏梅,孙秀玲,刘瑞华,等.三七总皂苷调控miRNA-466b抑制急性心梗大鼠心肌细胞凋亡的研究 [J].中国实验方剂学杂志,2015,21(22):6-10.Liu YM,Sun XL,Liu RH,et al.Panax notoginseng regulation of miRNA-466b to inhibit cardiomyocyte apoptosis in rats with acute myocardial infarction [J].Chin J Exp Tradit Med Formulae,2015,21(22):6-10.
[7] 郑茵红,高瑞兰,朱大元,等.三七总皂苷及其单体对人骨髓造血祖细胞增殖作用的研究 [J].中国中西医结合急救杂志,2003,10(3):135-137.Zheng YH,Gao RL,Zhu DY,et al.Proliferation of hematopoietic progenitor cells from human bone marrow induced by panax notoginosides [J].Chin J TCM WM Crit Care,2003,10(3):135-137.
[8] 杨军,任婷,李学东,等.三七总皂甙促进骨折愈合作用的实验研究 [J].医学研究杂志,2010,39(6):69-71.Yang J,Ren T,Li XD,et al.The experiment study of total panax notoginseng on promoting fracture healing [J].J Med Res,2010,39(6):69-71.
[9] 吴韶,张瑜,杨春红,等.三七不同部位中止血活血成分三七皂苷 Fc 的HPLC测定 [J].中草药,2016,47(18):3289-3292.Wu S,Zhang Y,Yang CH,et al.HPLC determination of notoginsenoside Fc,a hemostatic and activating blood component,in different parts of Panax notoginseng [J].Chin Tradit Herbal Drug,2016,47(18):3289-3292.
[10] Ning G,Liu X,Dai M,et al.MicroRNA-92a upholds Bmp signaling by targeting noggin3 during pharyngeal cartilage formation [J].Dev Cell,2013,24(3):283-295.
[11] 李响.Mta3/NuRD复合体在斑马鱼初级造血过程中的功能研究 [D].清华大学,2010:56-57.Li X.Functional analysis of Mta3/NuRD complex in zebrafish primitive hematopoiesis [D].Tsinghua University,2010:56-57.
[12] 陈磊,刘怡.模式生物斑马鱼在中药研究中的应用 [J].药学学报,2012,47(4):434-439.Chen L,Liu Y.Application of zebrafish model organism in the research of Chinese materia medica [J].Acta Pharm Sin,2012,47(4):434-439.
[13] 高瑞兰,徐卫红,林筱洁,等.三七皂甙对造血细胞GATA-1 和GATA-2 转录调控蛋白的诱导作用 [J].中华血液学杂志,2004,25(5):281-284.Gao RL,Xu WH,Lin XJ,et al.Up-regulation of transcription factors GATA-1 and GATA-2 induced by Panax notoginosides in hematopoietic cells [J].Chin J Hematol,2004,25(5):281-284.