Qingfei Paidu Decoction promotes differentiation of megakaryocytes and production of platelet

WAN Yuwei; WANG Peichun; GE Guangbo; LYU Chao; ZHAO Jing; ZHANG Weidong; LIU Xuan

doi:10.16306/j.1008-861x.2021.05.009

您当前的位置：

首页 >

文章列表页 >

Qingfei Paidu Decoction promotes differentiation of megakaryocytes and production of platelet

Experiment Research | 更新时间：2022-09-09

- Qingfei Paidu Decoction promotes differentiation of megakaryocytes and production of platelet
- Academic Journal of Shanghai University of Traditional Chinese Medicine Vol. 35, Issue 5, Pages: 52-60(2021)
- 作者机构：
  
  1.上海中医药大学交叉科学研究院（上海　201203）
  2.海军军医大学药学院（上海　200433）
- 作者简介：
- 基金信息：
- DOI：10.16306/j.1008-861x.2021.05.009
  CLC：
扫描看全文
WAN Yuwei, WANG Peichun, GE Guangbo, et al. Qingfei Paidu Decoction promotes differentiation of megakaryocytes and production of platelet. [J]. Academic Journal of Shanghai University of Traditional Chinese Medicine 35(5):52-60(2021)
DOI：

WAN Yuwei, WANG Peichun, GE Guangbo, et al. Qingfei Paidu Decoction promotes differentiation of megakaryocytes and production of platelet. [J]. Academic Journal of Shanghai University of Traditional Chinese Medicine 35(5):52-60(2021) DOI： 10.16306/j.1008-861x.2021.05.009.

摘要

目的：,2,考察清肺排毒汤（Qingfei Paidu Decoction，QFPDD）对巨核细胞分化和血小板生成的影响。,方法：,2,以人源巨核细胞DAMI细胞株为研究对象。制备QFPDD水煎液。①将DAMI细胞分为QFPDD不同浓度（0、25、50、75 mg/L）组，分别给予相应浓度的药物干预。药物作用48 h后，CCK-8法检测细胞增殖，流式细胞仪检测细胞DNA含量，PCR检测转录因子,aNF-E2,、,fNF-E2,、,GATA-1,和,Fli-1,的mRNA表达水平。②将细胞分为对照组和QFPDD不同浓度（25、50、75 mg/L）组，各组均给予1 μmol/L PMA和10 ng/ml TPO以诱导细胞分化，连续7 d；同时，QFPDD组给予相应浓度药液干预。干预7 d后，在无药物和诱导剂刺激条件下继续培养3 d，收集细胞和上清液。流式细胞仪检测细胞DNA含量、血小板样颗粒数目，并进行巨核细胞形态分析。,结果：,2,①不同浓度的QFPDD干预对未分化的DAMI细胞的增殖无明显影响。50、75 mg/L QFPDD干预能够诱导未分化的DAMI细胞进入G2/M期，使DNA含量为4N的细胞占比显著增加（,P,<,0.01）。②QFPDD干预可使诱导分化的DAMI细胞的DNA倍性显著增加，QFPDD 25、50、75 mg/L组中DNA倍性≥8N的细胞占比明显高于对照组（,P,<,0.05，,P,<,0.01），且各浓度组具有产生前血小板颗粒能力的细胞数量明显多于对照组（,P,<,0.01），各浓度组细胞培养基中血小板样颗粒数目均显著高于对照组（,P,<,0.01）。③QFPDD 75 mg/L组转录因子,fNF-E2,、,GATA-1,和,Fli-1,的mRNA表达量较对照组显著升高（,P,<,0.01），50 mg/L组,Fli-1, mRNA表达量亦显著升高（,P,<,0.01），但各浓度QFPDD作用对,aNF-E2, mRNA表达均无明显影响。,结论：,2,QFPDD能够促进DAMI细胞的分化和多倍体化，促进血小板样颗粒的生成，其作用机制可能与上调转录因子,fNF-E2,、,GATA-1,和,Fli-1,的表达相关。

Abstract

Objective:,2,To investigate the effects of Qingfei Paidu Decoction(QFPDD) on megakaryocyte differentiation and platelet production.,Methods:,2,Human megakaryocyte DAMI cell line was taken as the research object.The water decoction of QFPDD was prepared.①DAMI cells were divided into the QFPDD groups with different concentrations(0, 25, 50, 75 mg/L), and were treated with the decoction at corresponding concentration.After treatment for 48 hours, the cell proliferation was detected by CCK-8 assay, the DNA content was detected by flow cytometry, and the mRNA expression levels of transcription factors ,aNF-E,2,fNF-E2,GATA-1, and ,Fli-1, were detected by PCR.②The cells were divided into the control group and QFPDD groups with different concentrations(25, 50, 75 mg/L) . All groups were treated with 1 μmol/L PMA and 10 ng/ml TPO for 7 days, to induce cell differentiation; meanwhile, QFPDD intervention groups were treated with the decoction at corresponding concentration.After intervention for 7 days, the cells were cultured for 3 days without drug and inducer stimulation, and then the cells and supernatant were collected.The DNA content and the number of platelet-like particles were detected by flow cytometry, and the morphology of megakaryocytes was analyzed.,Results:,2,①QFPDD at different concentrations showed no significant effect on the proliferation of undifferentiated DAMI cells.QFPDD at concentration of 50 mg/L and 75 mg/L could induce the undifferentiated DAMI cells into G2/M phase, which significantly increased the proportion of cells with DNA content of 4N(,P,<,0.01) .②QFPDD treatment could significantly increase the DNA ploidy of differentiated DAMI cells.The proportion of cells with DNA ploidy≥8N in the QFPDD groups with different concentrations(25, 50 and 75 mg/L) was significantly higher than that in the control group(,P,<,0.05,P,<,0.01) .Moreover, the number of cells with the ability to produce proplatelet particles in all QFPDD treatment groups was significantly higher than that in the control group(,P,<,0.01), and the number of platelet-like particles in cell culture medium of all QFPDD treatment groups was significantly higher than that of the control group(,P,<,0.01) .③The mRNA expression levels of transcription factors ,fNF-E2,GATA-1, and ,Fli-1, in the QFPDD 75 mg/L group were significantly higher than those in the control group(,P,<,0.01), and the expression level of ,Fli-1, mRNA in the QFPDD 50 mg/L group was also significantly higher than that in the control group(,P,<,0.01), but QFPDD at different concentrations showed no significant effect on the expression of ,aNF-E2, mRNA.,Conclusion:,2,QFPDD can promote the differentiation and polyploidization of DAMI cells, as well as the production of platelet-like particles.Its mechanism may be related to up-regulating the expressions of transcription factors ,fNF-E2,GATA-1, and ,Fli-1,.

关键词

清肺排毒汤巨核细胞分化血小板

Keywords

Qingfei Paidu Decoctionmegakaryocytedifferentiationplatelet

references

GU S X，TYAGI T，JAIN K，et al. Thrombocytopathy and endotheliopathy： crucial contributors to COVID-19 thromboinflammation[J]. Nat Rev Cardiol，2021，18(3)： 194-209.

KHURRAM R，NAIDU V，BUTT M F，et al. Superior ophthalmic vein thrombosis secondary to COVID-19： an index case[J]. Radiol Case Rep，2021，16(5)： 1138-1143.

GOSHUA G，PINE A B，MEIZLISH M L，et al. Endotheliopathy in COVID-19-associated coagulopathy： evidence from a single-centre，cross-sectional study [J]. Lancet Haematol，2020，7(8)： e575-e582.

CÜRE E，CÜRE M C. COVID-19 May Predispose to Thrombosis by Affecting Both Vascular Endothelium and Platelets[J]. Clin Appl Thromb Hemost，2020，26： 1076029620933945. doi：10.1177/1076029620933945http://doi.org/ 10.1177/1076029620933945.

NICOLAI L，LEUNIG A，BRAMBS S，et al. Immunothrombotic Dysregulation in COVID-19 Pneumonia Is Associated With Respiratory Failure and Coagulopathy[J]. Circulation，2020，142(12)： 1176-1189.

AID M，BUSMAN-SAHAY K，VIDAL S J，et al. Vascular Disease and Thrombosis in SARS-CoV-2-Infected Rhesus Macaques [J]. Cell，2020，183(5)： 1354-1366.

YANG X，YANG Q，WANG Y，et al. Thrombocytopenia and its association with mortality in patients with COVID-19[J]. J Thromb Haemost，2020，18(6)： 1469-1472.

BOMHOF G，MUTSAERS P G，LEEBEEK F W，et al. COVID-19-associated immune thrombocytopenia[J]. Br J Haematol，2020，190(2)： e61-e64.

GREINACHER A，THIELE T，WARKENTIN T E，et al. Thrombotic Thrombocytopenia after ChAdOx1 nCov-19 Vaccination [J]. N Engl J Med，2021，384(22)： 2092-2101.

SCHULTZ N H，SØRVOLL I H，MICHELSEN A E，et al. Thrombosis and Thrombocytopenia after ChAdOx1 nCoV-19 Vaccination[J]. N Engl J Med，2021，384(22)： 2124-2130.

RAADSEN M，DU TOIT J，LANGERAK T，et al. Thrombocytopenia in Virus Infections[J]. J Clin Med，2021，10(4)： 877.

国家中医药管理局. 关于推荐在中西医结合救治新型冠状病毒感染的肺炎中使用“清肺排毒汤”的通知[EB/OL]. (2020-02-06). http://yzs.satcm.gov.cn/zhengcewenjian/2020-02-07/12876.htmlhttp://yzs.satcm.gov.cn/zhengcewenjian/2020-02-07/12876.html.

国家中医药管理局. 中医药有效方剂筛选研究取得阶段性进展试点省份临床观察显示：清肺排毒汤治疗总有效率可达90%以上[EB/OL]. (2020-02-06). http://bgs.satcm.gov.cn/gongzuodongtai/2020-02-06/12866.htmlhttp://bgs.satcm.gov.cn/gongzuodongtai/2020-02-06/12866.html.

SHI N，LIU B，LIANG N，et al. Association between early treatment with Qingfei Paidu decoction and favorable clinical outcomes in patients with COVID-19： A retrospective multicenter cohort study[J]. Pharmacol Res，2020，161： 105290.

LUO E，ZHANG D，LUO H，et al. Treatment efficacy analysis of traditional Chinese medicine for novel coronavirus pneumonia（COVID-19）： an empirical study from Wuhan，Hubei Province，China[J]. Chin Med，2020，15： 34.

ZHAO J，TIAN S，LU D，et al. Systems pharmacological study illustrates the immune regulation，anti-infection，anti-inflammation，and multi-organ protection mechanism of Qing-Fei-Pai-Du decoction in the treatment of COVID-19 [J]. Phytomedicine，2021，85： 153315.

LIU W，HUANG J，ZHANG F，et al. Comprehensive profiling and characterization of the absorbed components and metabolites in mice serum and tissues following oral administration of Qing-Fei-Pai-Du decoction by UHPLC-Q-Exactive-Orbitrap HRMS[J]. Chin J Nat Med，2021，19(4)： 305-320.

WAN Y W，LIU W，FENG M T，et al. LXRβ is involved in the control of platelet production from megakaryocytes[J]. Blood Cells Mol Dis，2021，89： 102568.

PATEL S R，HARTWIG J H，ITALIANO J E. The biogenesis of platelets from megakaryocyte proplatelets[J]. J Clin Invest，2005，115(12)： 3348-3354.

MACHLUS K R，ITALIANO J E. The incredible journey： From megakaryocyte development to platelet formation[J]. J Cell Biol，2013，201(6)： 785-796.

VAINCHENKER W，RASLOVA H. Megakaryocyte polyploidization： role in platelet production[J]. Platelets，2020，31(6)： 707-716.

LEV P R，GOETTE N P，GLEMBOTSKY A C，et al. Production of functional platelet-like particles by the megakaryoblastic DAMI cell line provides a model for platelet biogenesis[J]. Platelets，2011，22(1)： 28-38.

陈子兴. 巨核细胞发育和血小板生成的调控机理及巨核细胞和血小板的临床应用[J]. 中国病理生理杂志，2019，35(10)： 1901-1908.

GARCÍA P，BERLANGA O，VEGIOPOULOS A，et al. c-Myb and GATA-1 alternate dominant roles during megakaryocyte differentiation[J]. J Thromb Haemost，2011，9(8)： 1572-1581.

GASIOREK J J，BLANK V. Regulation and function of the NFE2 transcription factor in hematopoietic and non-hematopoietic cells [J]. Cell Mol Life Sci，2015，72(12)： 2323-2335.

PISCHEDDA C，COCCO S，MELIS A，et al. Isolation of a differentially regulated splicing isoform of human NF-E2[J]. Proc Natl Acad Sci USA，1995，92(8)： 3511-3515.

TOKI T，ARAI K，TERUI K，et al. Functional characterization of the two alternative promoters of human p45 NF-E2 gene[J]. Exp Hematol，2000，28(10)： 1113-1119.

EISBACHER M，HOLMES M L，NEWTON A，et al. Protein-protein interaction between Fli-1 and GATA-1 mediates synergistic expression of megakaryocyte-specific genes through cooperative DNA binding[J]. Mol Cell Biol，2003，23(10)： 3427-3441.

ZANG C，LUYTEN A，CHEN J，et al. NF-E2，FLI1 and RUNX1 collaborate at areas of dynamic chromatin to activate transcription in mature mouse megakaryocytes[J]. Sci Rep，2016，6： 30255.

FOCK E L，YAN F，PAN S，et al. NF-E2-mediated enhancement of megakaryocytic differentiation and platelet production in vitro and in vivo[J]. Exp Hematol，2008，36(1)： 78-92.

SUN X，ZHAO Y N，QIAN S，et al. Ginseng-Derived Panaxadiol Saponins Promote Hematopoiesis Recovery in Cyclophosphamide-Induced Myelosuppressive Mice： Potential Novel Treatment of Chemotherapy-Induced Cytopenias[J]. Chin J Integr Med，2018，24(3)： 200-206.

LEVAY K，SLEPAK V Z. Tescalcin is an essential factor in megakaryocytic differentiation associated with Ets family gene expression[J]. J Clin Invest，2007，117(9)： 2672-2683.

Views

488

下载量

CSCD

CNKI被引量

Alert me when the article has been cited

提交

Tools

Publicity Resources

Coagulopathy in patients with COVID⁃19 and its possible mechanisms as well as treatment strategies

Related Author

No data

Related Institution

Institute of Interdisciplinary Integrative Medicine Research， Shanghai University of Traditional Chinese Medicine

⁰