黄芪甲苷通过诱导M1型巨噬细胞极化发挥抗肿瘤作用的机制

王莉新; 吴文斌; 胥孜杭; 焦肖宁; 苏琳; 朱杨壮壮; 陈晓; 邹纯朴; 朱诗国

doi:10.13422/j.cnki.syfjx.20190822

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黄芪甲苷通过诱导M1型巨噬细胞极化发挥抗肿瘤作用的机制

中医药复方及其有效成分抗肺癌作用研究专题 | 更新时间：2021-02-09

- 黄芪甲苷通过诱导M1型巨噬细胞极化发挥抗肿瘤作用的机制
- Anti-tumor Effect of Astragaloside by Inducing M1 Macrophage Polarization
- 中国实验方剂学杂志 2019年25卷第14期页码：19-24
- 作者机构：
  
  上海中医药大学，上海　 201203
- 作者简介：
  
  王莉新，博士，助理研究员，从事中西医结合抗肿瘤研究，E-mail：wlx831208@126.com
  朱诗国，博士，研究员，从事中西医结合抗肿瘤研究， E-mail：jusco105@163.com
- 基金信息：
  
  国家自然科学基金项目(81803933;81473237);上海市卫生和计划生育委员会科研课题项目(20154Y0167)
- DOI：10.13422/j.cnki.syfjx.20190822
  中图分类号： R22; R242; R2-031; R285.5;R392.5; R273
- 收稿日期：2018-10-08，
  
  网络出版日期：2019-01-04，
  
  纸质出版日期：2019-07-20
- 稿件说明：
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王莉新, 吴文斌, 胥孜杭, 等. 黄芪甲苷通过诱导M1型巨噬细胞极化发挥抗肿瘤作用的机制[J]. 中国实验方剂学杂志, 2019,25(14):19-24.

Li-xin WANG, Wen-bin WU, Zi-hang XU, et al. Anti-tumor Effect of Astragaloside by Inducing M1 Macrophage Polarization[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(14): 19-24.
王莉新, 吴文斌, 胥孜杭, 等. 黄芪甲苷通过诱导M1型巨噬细胞极化发挥抗肿瘤作用的机制[J]. 中国实验方剂学杂志, 2019,25(14):19-24. DOI： 10.13422/j.cnki.syfjx.20190822.

Li-xin WANG, Wen-bin WU, Zi-hang XU, et al. Anti-tumor Effect of Astragaloside by Inducing M1 Macrophage Polarization[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(14): 19-24. DOI： 10.13422/j.cnki.syfjx.20190822.

摘要

目的：

探讨黄芪甲苷对巨噬细胞极化的影响及其可能的抗肿瘤免疫机制。

方法：

通过噻唑兰(methylthiazolyldiphenyl-tetrazolium bromide，MTT)比色法检测不同浓度黄芪甲苷作用不同时间，对巨噬细胞的细胞毒作用，以选取合适的黄芪甲苷作用浓度；将巨噬细胞与肿瘤细胞以1∶1进行共孵育，并用0.1 mg·L

－1

黄芪甲苷作用24 h，通过虫荧光素酶(luciferase，Luc)杀伤实验检测巨噬细胞对肿瘤细胞的杀伤效率；以0.1 mg·L

－1

黄芪甲苷作用巨噬细胞24 h，通过流式细胞术检测巨噬细胞CD16/32，CD206表达，通过实时荧光定量聚合酶链式反应(Real-time PCR)检测巨噬细胞诱导型一氧化氮合酶(inducible nitric oxide synthase，iNOS)，精氨酸-1(Arginine-1，Arg-1)及细胞因子白细胞介素-1

(interleukin-1

，IL-1

)，肿瘤坏死因子-

(tumor necrosis factor-

，TNF-

)，白细胞介素-12(interleukin-12，IL-12)，白细胞介素-10(interleukin-10，IL-10)，转化生长因子-

(transforming growth factor-

，TGF-

)mRNA表达，通过蛋白免疫印迹法(Western blot)检测巨噬细胞信号传导及转录激活因子1(Signal transducers and activators of transcription 1

STAT1)，磷酸化信号传导及转录激活因子1(phosphorylation signal transducers and activators of transcription 1

p-STAT1)表达。

结果：

当黄芪甲苷质量浓度为0.1 mg·L

－1

时，对巨噬细胞无细胞毒作用。与空白组比较，Luc杀伤实验结果显示黄芪甲苷可以显著促进巨噬细胞介导的肿瘤杀伤；Real-time PCR结果显示黄芪甲苷可以诱导巨噬细胞表面M1型巨噬细胞标志CD16/32表达增高，促进M1型巨噬细胞特异性指标iNOS mRNA表达，同时诱导M1型巨噬细胞相关细胞因子IL-1

，TNF-

，IL-12 mRNA表达增高；Western blot结果显示黄芪甲苷可以促进巨噬细胞STAT1发生磷酸化。

结论：

黄芪甲苷可以通过促进巨噬细胞内STAT1发生磷酸化，进而诱导巨噬细胞向M1型巨噬细胞发生极化，并启动巨噬细胞相关的抗肿瘤免疫应答。

Abstract

Objective:

To investigate the effect of astragaloside on the macrophage polarization and the possible anti-tumor immunity mechanism of astragaloside.

Method:

The cytotoxic effect of different concentrations of astragaloside at different time points on macrophage was measured by methylthiazolyldiphenyl-tetrazolium bromide (MTT)

in order to choose the suitable concentration of astragaloside

macrophages were co-cultured with tumor cells at the ratio 1∶1

and the effect of astragaloside on macrophage-mediated lysis of tumor cells was performed by biophotonic cytotoxicity assay after the mixed cells were effected with 0.1 mg·L

-1

astragaloside for 24 h. Macrophages were dealt with 0.1 mg·L

-1

astragaloside for 24h

the expressions of CD16/32 and CD206 in macrophages were performed by flow cytometry

the mRNA expressions of macrophage inducible nitric oxide synthase (iNOS)

Arginine-1 (Arg-1)

interleukin-1

(IL-1

)

tumor necrosis factor-

(TNF-

)

interleukin-12 (IL-12)

interleukin-10 (IL-10) and transforming growth factor-

(TGF-

) were measured by Real-time PCR

the protein expressions of macrophage signal transducers and activators of transcription 1 (STAT1) and phosphorylation signal transducers and activators of transcription 1 (p-STAT1) were determined by Western blot.

Result:

Astragaloside had no effect on the viability of macrophages with 0.1 mg·L

-1

. Compared with control group

astragaloside obviously enhanced the macrophage-mediated lysis of tumor cells according to the biophotonic cytotoxicity assay

induced the M1 macrophage marker CD16/32 expression according to flow cytometry

increased the mRNA expressions of iNOS

IL-1

TNF-

and IL-12 according to the Real-time PCR

and promoted the phosphorylation of STAT1 in macrophages on the basis of Western blot.

Conclusion:

Astragaloside could induce M1 macrophage polarization by increasing the phosphorylation of STAT1

and initiate macrophage-related anti-tumor immunity response.

关键词

Keywords

references

CHEN W , ZHENG R , Baade P D , et al . Cancer statistics in China, 2015 [J]. CA Cancer J Clin , 2016 , 66 ( 2 ): 115 - 132 .

Siegel R , Naishadham D , Jemal A . Cancer statistics, 2013 [J]. CA Cancer J Clin , 2013 , 63 ( 1 ): 11 - 30 .

Albini A , Magnani E , Noonan D M . The tumor microenvironment: biology of a complex cellular and tissue society [J]. Q J Nucl Med Mol Imaging , 2010 , 54 ( 3 ): 244 - 248 .

Schreiber R D , Old L J , Smyth M J . Cancer immunoediting: integrating immunity's roles in cancer suppression and promotion [J]. Science , 2011 , 331 ( 6024 ): 1565 - 1570 .

Kerkar S P , Restifo N P . Cellular constituents of immune escape within the tumor microenvironment [J]. Cancer Res , 2012 , 72 ( 13 ): 3125 - 3130 .

YUAN R , LI S , GENG H , et al . Reversing the polarization of tumor-associated macrophages inhibits tumor matastasis [J]. Int Immunopharmacol , 2017 , 49 : 30 - 37 .

LUO Y , WU J , ZHU X , et al . NK cell-dependent growth inhibition of Lewis lung cancer by Yu-Ping-Feng, an ancient Chinese herbal formula [J]. Med Inflamm , 2016 , doi: 10.1155/2016/3541283 http://doi.org/10.1155/2016/3541283 .

陈卓，于卫江．注射用黄芪多糖对非小细胞肺癌放疗的作用及对机体免疫的影响 [J]．中国实验方剂学杂志， 2013 ， 19 ( 6 )： 309 - 313 ．

刘苓霜．刘嘉湘辨治肺癌经验 [J]．中医文献杂志， 2006 ， 24 ( 2 )： 38 - 40 ．

刘嘉湘，陈树森，郁仁存，等．肺癌证治 [J]．中医杂志， 1986 ， 27 ( 3 )： 4 - 7 ．

桑国优，韦世秀，刘成军．黄芪抗肿瘤作用机制和临床应用研究进展 [J]．时珍国医国药， 2008 ， 19 ( 12 )： 3032 - 3034 ．

杨苏钰，唐德才，曹子丰，等．黄芪甲苷配伍姜黄素对人卵巢癌HO-8910原位移植瘤转移的抑瘤作用 [J]．中国实验方剂学杂志， 2017 ， 23 ( 6 )： 155 - 160 ．

Sica A , Mantovani A . Macrophage plasticity and polarization: in vivo veritas [J]. J Clin Invest , 2012 , 122 ( 3 ): 787 - 795 .

Mosser D M , Edwards J P . Exploring the full spectrum of macrophage activation [J]. Nat Rev Immunol , 2008 , 8 ( 12 ): 958 - 969 .

Lewis C E , Pollard J W . Distinct role of macrophages in different tumor microenvironments [J]. Cancer Res , 2006 , 66 ( 2 ): 605 - 612 .

Mantovani A , Sozzani S , Locati M , et al . Macrophage polarization: tumor-associated macrophages as a paradigm for polarized M2 mononuclear phagocytes [J]. Trends Immunol , 2002 , 23 ( 11 ): 549 - 555 .

Goerdt S , Orfanos C E . Other functions, other genes: alternative activation of antigen-presenting cells [J]. Immunity , 1999 , 10 ( 2 ): 137 - 142 .

Mantovani A , Allavena P . The interaction of anticancer therapies with tumor-associated macrophage [J]. J Exp Med , 2015 , 212 ( 4 ): 435 - 445 .

Mantovani A , Marchesi F , Malesci A , et al . Tumor-associated macrophages as treatment targets in oncology [J]. Nat Rev Clin Oncol , 2017 , 14 ( 7 ): 399 - 416 .

Biswas S K , Allavena P , Mantovani A . Tumor-associated macrophage: functional diversity, clinical significance, and open questions [J]. Semin Immunopathol , 2013 , 35 ( 5 ): 585 - 600 .

Hagemann T , Lawrence T , McNeish I , et al . Re-educating tumor-associated macrophages by targeting NF-kappaB [J]. J Exp Med , 2008 , 205 ( 6 ): 1261 - 1268 .

KE S , Ziemiecki A , Wilks A F , et al . Polypeptide signaling to the nucleus through tyrosine phosphorylation of Jak and Stat proteins [J]. Nature , 1993 , 366 ( 6455 ): 580 - 583 .

Darnell J E J , Kerr I M , Stark G R . Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins [J]. Science , 1994 , 264 ( 5164 ): 1415 - 1421 .

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