Renshen Yangrongtang Alleviating Myelosuppression by Reducing Neutrophil Extracellular Traps Through Regulating ROS/MPO

ZHANG Jing; LIU Rongxing; ZENG Jinhao; NIAN Qing

doi:10.13422/j.cnki.syfjx.20241722

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Renshen Yangrongtang Alleviating Myelosuppression by Reducing Neutrophil Extracellular Traps Through Regulating ROS/MPO

更新时间：2025-03-04

- Renshen Yangrongtang Alleviating Myelosuppression by Reducing Neutrophil Extracellular Traps Through Regulating ROS/MPO
  增强出版
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 31, Issue 6, Pages: 39-46(2025)
- 作者机构：
  
  1.四川省医学科学院四川省人民医院（电子科技大学附属医院），成都 610072
  2.陆军军医大学第二附属医院，重庆 400037
  3.成都中医药大学附属医院，成都 610072
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20241722
  CLC： R242;R551.3;R285
- Received：17 May 2024，
  
  Accepted：12 July 2024，
  
  Online First：29 July 2024，
  
  Published：20 March 2025
- 稿件说明：
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张靖,刘荣兴,曾进浩等.人参养荣汤通过调控ROS/MPO减少中性粒细胞胞外诱捕网缓解骨髓抑制[J].中国实验方剂学杂志,2025,31(06):39-46.

ZHANG Jing,LIU Rongxing,ZENG Jinhao,et al.Renshen Yangrongtang Alleviating Myelosuppression by Reducing Neutrophil Extracellular Traps Through Regulating ROS/MPO[J].Chinese Journal of Experimental Traditional Medical Formulae,2025,31(06):39-46.
张靖,刘荣兴,曾进浩等.人参养荣汤通过调控ROS/MPO减少中性粒细胞胞外诱捕网缓解骨髓抑制[J].中国实验方剂学杂志,2025,31(06):39-46. DOI： 10.13422/j.cnki.syfjx.20241722.

ZHANG Jing,LIU Rongxing,ZENG Jinhao,et al.Renshen Yangrongtang Alleviating Myelosuppression by Reducing Neutrophil Extracellular Traps Through Regulating ROS/MPO[J].Chinese Journal of Experimental Traditional Medical Formulae,2025,31(06):39-46. DOI： 10.13422/j.cnki.syfjx.20241722.

摘要

目的

探讨人参养荣汤调节活性氧（ROS）、髓过氧化物酶（MPO）及中性粒细胞胞外诱捕网（NETs）的表达从而缓解骨髓抑制的潜在机制。

方法

将K562细胞分为空白组，依托泊苷组（40 μmol·L

-1

），依托泊苷+人参养荣汤（冻干粉）低、中、高质量浓度组（2、4、8 g·L

-1

）。液相色谱-质谱联用技术（LC-MS）检测人参养荣汤冻干粉成分；酶联免疫吸附测定法（ELISA）检测各组ROS、MPO及NETs表达；蛋白免疫印迹法（Western blot）检测胞内MPO和中性粒细胞弹性蛋白酶（NE）表达。将20只8周龄雄性小鼠随机分为正常组，依托泊苷组（100 mg·kg

-1

），依托泊苷+人参养荣汤低、中、高剂量组（0.1、0.5、2.0 g·kg

-1

），除空白组常温磷酸盐缓冲液（PBS）灌胃，依托泊苷组腹腔注射3 d外，剩余组依托泊苷给药3 d后人参养荣汤连续灌胃14 d。血常规分析仪检测小鼠外周血象相关指标；Western blot检测小鼠骨髓细胞内MPO及NE表达改变；ELISA检测小鼠

骨髓细胞ROS，MPO，NETs改变；小鼠股骨行MPO、NE免疫组化染色；电镜扫描分析小鼠骨髓细胞中NETs结构变化。

结果

LC-MS结果显示人参养荣汤冻干粉中含有当归、黄芪、人参等完整的原药成分；K562细胞中，与依托泊苷组比较，ELISA结果表明依托泊苷+人参养荣汤高、中质量浓度组MPO、ROS及NETs水平均降低（

0.05，

0.01），同时Western blot显示高质量浓度组明显降低K562细胞内MPO、NE蛋白表达（

0.05，

0.01）。小鼠体内，与依托泊苷组比较，依托泊苷+人参养荣汤高质量浓度组红细胞（RBC）、白细胞（WBC）及血小板（PLT）数量上升（

0.05）；ELISA结果提示依托泊苷+人参养荣汤低、中、高质量浓度组小鼠体内ROS、MPO、NETs浓度降低（

0.05，

0.01）；Western blot结果说明依托泊苷+人参养荣汤低、中、高质量浓度组小鼠骨髓细胞内MPO、NE表达较依托泊苷组降低（

0.05，

0.01）；扫描电镜观察人参养荣汤可减少依托泊苷作用后小鼠骨髓细胞内NETs结构生成。

结论

人参养荣汤通过抑制ROS/MPO，减少细胞内NETs形成，从而缓解依托泊苷诱导的骨髓抑制。

Abstract

Objective

To investigate the potential mechanism of Renshen Yangrongtang in alleviating myelosuppression by regulating the expression of reactive oxygen species （ROS）， myeloperoxidase （MPO）， and neutrophil extracellular traps （NETs）.

Methods

K562 cells were divided into blank group， etoposide group （40 μmol·L

-1

）， and etoposide+Renshen Yangrongtang freeze-dried powder groups with low-， medium-， and high-dose （2， 4， 8 g·L

-1

）. Liquid chromatography-mass spectrometry （LC-MS） was used to determine the freeze-dried powder of Renshen Yangrongtang. Enzyme-Linked Immunosorbent Assay （ELISA） was used to detect ROS， MPO， and NETs expression in each group. Western blot analysis was performed to assess intracellular MPO and NE expressions. Twenty 8-week-old male mice were randomly divided into blank group， etoposide group （100 mg·kg

-1

）， and etoposide + Renshen Yangrongtang groups with low-， medium-， and high-dose （0.1， 0.5， 2.0 g·kg

-1

）. Except for the blank group that received PBS via gavage at room temperature， and the etoposide group that received an intraperitoneal injection for 3 days， the re

maining groups received gavage of Renshen Yangrongtang for 14 consecutive days after 3 days of etoposide administration. The peripheral blood related indicators were detected through an automated hematology analyzer； Western blot analysis was performed to assess MPO and neutrophil elastase （NE） expression changes in the marrow cells of mice. Enzyme-linked immunosorbent assay （ELISA） was used to detect ROS， MPO， and NETs changes in the marrow cells of mice. MPO and NE on femur bones were stained through immunohistochemistry. Scanning electron microscopy was used to analyze the structural changes of NETs in the marrow cells of mice after drug administration.

Results

LC-MS results showed that the freeze-dried powder of Renshen Yangrongtang contained complete technical materials such as Chinese angelica，

Astragalus mongholicus

， and ginseng. In K562 cells， compared with the etoposide group， ELISA results indicated that the concentrations of MPO， ROS， and NETs in the etoposide + Renshen Yangrongtang medium and high-dose groups were decreased （

0.05，

0.01）， and Western blot data showed that the etoposide high-dose group significantly reduced the expression of MPO and NE protein in K562 cells （

0.05，

0.01）.

In vivo

， compared with the etoposide group， the number of RBC， WBC， and PLT in the etoposide+Renshen Yangrongtang high-dose group increased significantly （

0.05）. ELISA results suggested that in the etoposide+Renshen Yangrongtang low-， medium-， and high-dose groups， the concentration of mice ROS， MPO， and NETs significantly decreased （

0.05，

0.01）. Western blot results revealed that compared with the etoposide group， the expressions of MPO and NE in the marrow cells of mice in the etoposide + Renshen Yangrongtang low-， medium- and high-dose groups were significantly decreased （

0.05，

0.01）. Scanning electron microscopy observations revealed that Renshen Yangrongtang reduced the NETs structure generation in the marrow cells of mice after the influence of etoposide.

Conclusion

Renshen Yangrongtang can alleviate etoposide-induced myelosuppression by inhibiting ROS/MPO and reducing the formation of intracellular NETs.

关键词

Keywords

references

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