清肺排毒汤促进巨核细胞分化和血小板生成的作用研究

万雨薇; 王沛纯; 葛广波; 吕超; 赵静; 张卫东; 刘璇

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

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清肺排毒汤促进巨核细胞分化和血小板生成的作用研究

实验研究 | 更新时间：2022-09-09

- 清肺排毒汤促进巨核细胞分化和血小板生成的作用研究
- Qingfei Paidu Decoction promotes differentiation of megakaryocytes and production of platelet
- 上海中医药大学学报 2021年35卷第5期页码：52-60
- 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：
  
  万雨薇，女，在读硕士生，主要从事中药药理学研究。王沛纯，女，在读硕士生，主要从事中药药理学研究（本文贡献与第一作者等同）
  张卫东，教授，博士生导师；E-mail：wdzhangy@hotmail.com。
  刘璇，研究员，博士生导师；E-mail：xuanliu@shutcm.edu.cn
- 基金信息：
  
  国家重点研发计划应对新冠肺炎疫情国际合作项目(2021YFE0200900);国家重点研发计划项目(2020YFC0845400);上海市科技创新行动计划生物医药科技支撑专项项目(2020XGKY10)
- DOI：10.16306/j.1008-861x.2021.05.009
  中图分类号：
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万雨薇, 王沛纯, 葛广波, 等. 清肺排毒汤促进巨核细胞分化和血小板生成的作用研究[J]. 上海中医药大学学报, 2021,35(5):52-60.

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, 2021,35(5):52-60.
万雨薇, 王沛纯, 葛广波, 等. 清肺排毒汤促进巨核细胞分化和血小板生成的作用研究[J]. 上海中医药大学学报, 2021,35(5):52-60. DOI： 10.16306/j.1008-861x.2021.05.009.

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, 2021,35(5):52-60. 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

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