中草药

通关藤注射液调节巨噬细胞极化协同多柔比星抗三阴性乳腺癌的作用机制

PDF下载

魏诗婷, 杨柳青, 陈小菲, 韩博洋, 郑亚娟, 张明亮, 吴娅丽, 张辉, 王晓艳, 李伟霞, 唐进法

中草药 | 药理与临床 2026,57(9): 3414-3428

收起

中草药 |药理与临床 2026 , 57 (9) : 3414 -3428

通关藤注射液调节巨噬细胞极化协同多柔比星抗三阴性乳腺癌的作用机制

全屏

魏诗婷, 杨柳青, 陈小菲, 韩博洋, 郑亚娟, 张明亮, 吴娅丽, 张辉, 王晓艳, 李伟霞, 唐进法

作者信息

通讯作者:

杨柳青

作者简介:

魏诗婷: 魏诗婷,硕士研究生,从事中药质量评价与合理用药研究。E-mail:wst17836939909@163.com

Mechanism of Tongguanteng Injection in regulating macrophage polarization and cooperating with doxorubicin in anti-triple negative breast cancer

WEI Shiting, YANG Liuqing, CHEN Xiaofei, HAN Boyang, ZHENG Yajuan, ZHANG Mingliang, WU Yali, ZHANG Hui, WANG Xiaoyan, LI Weixia, TANG Jinfa

Affiliations

doi: 10.7501/j.issn.0253-2670.2026.09.014

文章导航

摘要

收起

目的探究通关藤注射液（Tongguanteng Injection，TGT）调节巨噬细胞极化进而协同多柔比星（doxorubicin，DOX）抑制三阴性乳腺癌（triple-negative breast cancer，TNBC）生长的作用机制。方法构建4T1乳腺癌荷瘤小鼠模型，随机分为模型组、TGT组、DOX组及TGT联合DOX治疗组，观察肿瘤体积及质量，评估体内抗肿瘤疗效；采用苏木素-伊红（hematoxylin-eosin，HE）、TUNEL及免疫组化染色检测肿瘤组织病理、细胞凋亡及剪切型半胱氨酸天冬氨酸蛋白酶-3（cleaved cystein-asparate protease-3，cleaved Caspase-3）、B细胞淋巴瘤-2（B-cell lymphoma-2，Bcl-2）、Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）表达；通过免疫器官指数、血常规分析评估小鼠免疫功能；ELISA、qRT-PCR及免疫荧光染色检测肿瘤组织炎症因子水平和巨噬细胞极化标志物CD86、诱导型一氧化氮合酶（inducible nitric oxide synthase，iNOS）、CD206、精氨酸酶-1（arginase-1，Arg-1）的表达；流式细胞术及免疫组化分析肿瘤组织CD4⁺ T、CD8⁺ T细胞浸润比例；qRT-PCR检测T细胞耗竭相关因子及细胞毒性因子的mRNA表达；转录组测序及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）富集分析挖掘核心信号通路并进行验证。结果与模型组比较，TGT组、DOX组及TGT联合DOX治疗组小鼠的肿瘤体积和质量显著降低（P＜0.01、0.001），肿瘤组织出现不同程度坏死，肿瘤细胞凋亡率显著升高（P＜0.001），其中TGT联合DOX治疗组上述变化更为显著（P＜0.05、0.01、0.001）。与模型组相比，TGT组的促炎因子肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-1β（interleukin-1β，IL-1β）表达水平显著升高（P＜0.01、0.001），抗炎因子IL-4、IL-10表达水平显著降低（P＜0.05、0.01）；M1型巨噬细胞（CD86⁺/F4/80⁺）比例增加（P＜0.001），标志物CD86、iNOS mRNA表达水平显著升高（P＜0.05、0.01）；M2型巨噬细胞（CD206⁺/F4/80⁺）比例减少（P＜0.001），标志物CD206、Arg-1 mRNA表达水平显著降低（P＜0.01、0.001）；CD8⁺ T细胞比例显著升高（P＜0.05、0.001），CD4⁺ T细胞比例显著降低（P＜0.01）；T细胞耗竭指标程序性死亡受体-1（programmed cell death protein 1，PD-1）、T细胞免疫球蛋白黏蛋白分子-3（T cell immunoglobulin and mucin domain-containing protein-3，Tim-3）、淋巴细胞活化基因-3（lymphocyte activation gene-3，Lag-3）mRNA表达水平显著下调（P＜0.05、0.01），T细胞毒性因子穿孔素（perforin）、γ干扰素（interferon γ，IFN-γ）、颗粒酶B（granzyme B）mRNA表达水平显著上调（P＜0.05、0.001）；TGT联合DOX治疗组上述变化更加显著（P＜0.05、0.01、0.001）。转录组及KEGG富集分析显示，与模型组比较，联合治疗组差异基因显著富集于Janus激酶（Janus kinase，JAK）-信号转导和转录激活蛋白（signal transducer and activator of transcription，STAT）信号通路；免疫组化染色结果表明TGT联合DOX显著下调肿瘤细胞中p-JAK2和p-STAT3蛋白的表达（P＜0.001）。结论 TGT联合DOX显著抑制TNBC的生长，促进肿瘤相关巨噬细胞由M2型向M1型极化，促进CD8⁺ T细胞浸润并减轻其耗竭，其作用机制与抑制JAK-STAT信号通路的磷酸化有关。

关键词

通关藤注射液 / 多柔比星 / 三阴性乳腺癌 / 巨噬细胞极化 / 免疫调节 / JAK-STAT信号通路

Abstract

收起

Objective To investigate the mechanism by which Tongguanteng Injection (通关藤注射液, TGT) modulates macrophage polarization to synergize with doxorubicin (DOX) in inhibiting triple-negative breast cancer (TNBC) growth. Methods A 4T1 TNBC- bearing mouse model was constructed, randomly divided into model group, TGT group, DOX group and TGT combined with DOX treatment group. Tumor volume and weight were observed to evaluate in vivo antitumor efficacy. Hematoxylin-eosin (HE), TUNEL and immunohistochemical staining were employed to examine histopathological morphological changes in tumor tissues, tumor cell apoptosis, and protein expressions of cleaved cystein-asparate protease-3 (Caspase-3), B-cell lymphoma-2 (Bcl-2), and Bcl-2 associated X protein (Bax). Immune organ indices and routine blood analysis were conducted to assess immune function in mice. ELISA, qRT-PCR and immunofluorescence staining were performed to measure inflammatory cytokine levels and expressions of macrophage polarization markers CD86, inducible nitric oxide synthase (iNOS), CD206, arginase-1 (Arg-1) in tumor tissues. Flow cytometry and immunohistochemical analysis were used to determine the infiltration ratios of CD4⁺ T and CD8⁺ T cells, while qRT-PCR was employed to detect mRNA expression levels of T cell exhaustion-related factors and cytotoxic factors. Transcriptome sequencing and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis were conducted to identify core signaling pathways and perform validation. Results Compared with model group, the tumor volume and weight were significantly reduced in TGT group, DOX group and TGT combined with DOX treatment group (P < 0.01, 0.001), varying degrees of necrosis were observed in tumor tissues, the apoptosis rate of tumor cells was significantly increased (P < 0.001), these changes were more pronounced in TGT + DOX combination therapy group (P < 0.05, 0.01, 0.001). Compared with model group, the expression levels of pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were significantly increased in TGT group (P < 0.01, 0.001), while the expression levels of anti-inflammatory cytokines IL-4 and IL-10 were significantly decreased (P < 0.05, 0.01). The proportion of M1-type macrophages (CD86⁺/F4/80⁺) was increased (P < 0.001), with significantly elevated mRNA expression levels of markers CD86 and iNOS (P < 0.05, 0.01). The proportion of M2-type macrophages (CD206⁺/F4/80⁺) was decreased (P < 0.001), with significantly reduced mRNA expression levels of markers CD206 and Arg-1 (P < 0.01, 0.001). The proportion of CD8⁺ T cells was significantly increased (P < 0.05, 0.001), while the proportion of CD4⁺ T cells was significantly decreased (P < 0.01). The mRNA expression levels of T cell exhaustion markers programmed cell death protein-1 (PD-1), T cell immunoglobulin and mucin domain-containing protein-3 (Tim-3) and lymphocyte activation gene-3 (Lag-3) were significantly downregulated (P < 0.05, 0.01), whereas the mRNA expression levels of T cell cytotoxic factors perforin, interferon-γ (IFN-γ) and granzyme B were significantly upregulated (P < 0.05, 0.001). These changes were more pronounced in TGT combined with DOX treatment group (P < 0.05, 0.01, 0.001). Transcriptome and KEGG enrichment analyses revealed that, compared with model group, differentially expressed genes in the combination therapy group were significantly enriched in Janus kinase (JAK)-signal transducer and activator of transcription (STAT) signaling pathway. Immunohistochemical staining results indicated that TGT combined with DOX significantly downregulated the expressions of p-JAK2 and p-STAT3 proteins in tumor cells (P < 0.001). Conclusion TGT combined with DOX significantly inhibits the growth of TNBC, promotes the polarization of tumor associated macrophages from M2 to M1, promotes CD8⁺ T cell infiltration and reduces their exhaustion. Its mechanism is related to the inhibition of phosphorylation of JAK-STAT signaling pathway.

Key words

Tongguanteng Injection / doxorubicin / triple-negative breast cancer / macrophage polarization / immunomodulation / JAK-STAT signaling pathway

引用本文

魏诗婷, 杨柳青, 陈小菲, 韩博洋, 郑亚娟, 张明亮, 吴娅丽, 张辉, 王晓艳, 李伟霞, 唐进法. 通关藤注射液调节巨噬细胞极化协同多柔比星抗三阴性乳腺癌的作用机制. 中草药, 2026 , 57 (9) : 3414 -3428 . DOI: 10.7501/j.issn.0253-2670.2026.09.014

WEI Shiting, YANG Liuqing, CHEN Xiaofei, HAN Boyang, ZHENG Yajuan, ZHANG Mingliang, WU Yali, ZHANG Hui, WANG Xiaoyan, LI Weixia, TANG Jinfa. Mechanism of Tongguanteng Injection in regulating macrophage polarization and cooperating with doxorubicin in anti-triple negative breast cancer[J]. Chinese Traditional and Herbal Drugs, 2026 , 57 (9) : 3414 -3428 . DOI: 10.7501/j.issn.0253-2670.2026.09.014

基金

收起

广东省中医药局科研项目(20241084);广东省基础与应用基础研究基金项目(2021A1515110800)

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

张启越,燕德悦,陆潇曼,等.影像组学预测乳腺癌肿瘤浸润淋巴细胞的研究进展[J].磁共振成像, 2025,16(12):205-211.
马琴,王玉.放射治疗联合免疫治疗在三阴性乳腺癌中的研究进展[J].中国医学前沿杂志:电子版, 2025,17(5):136.
吴进敏,戚宇航,方静怡,等. FTO通过Wnt/β-catenin信号通路调节三阴性乳腺癌对阿霉素的耐药性[J].中国药理学通报, 2025, 41(12):2334-2341.
陈欣,郑琦,席玉棚,等.从五脏气探讨调气解毒法联合免疫检查点抑制剂在晚期非小细胞肺癌中的应用[J].中国医药导报, 2025, 22(12):136-140.
Zhou H R, Zhang M X, Cao H H, et al. Research progress on the synergistic anti-tumor effect of natural anti-tumor components of Chinese herbal medicine combined with chemotherapy drugs[J]. Pharmaceuticals, 2023, 16(12):1734.
邹蔓姝,雷昌,黄丹,等.用于肿瘤治疗的智能调控/响应肿瘤微环境的纳米递药系统研究进展[J].中国新药杂志, 2024, 33(15):1564-1570.
Waibl Polania J, Hoyt-Miggelbrink A, Tomaszewski W H,et al. Antigen presentation by tumor-associated macrophages drives T cells from a progenitor exhaustion state to terminal exhaustion[J]. Immunity, 2025, 58(1):232-246.
Wang K C, Liu W, Xu Q F, et al. Tenacissoside G synergistically potentiates inhibitory effects of 5-fluorouracil to human colorectal cancer[J].Phytomedicine, 2021, 86:153553.
Ma X Z, Xiao L L, Liu L T, et al. CD36-mediated ferroptosis dampens intratumoral CD8+T cell effector function and impairs their antitumor ability[J]. Cell Metab, 2021, 33(5):1001-1012.
吴子薇,李晓波,麻文谦,等.通关藤注射液研究进展[J].上海中医药大学学报, 2023, 37(4):71-82.
盛佳钰,张馨月,李琼,等.基于肿瘤微环境中TLR4/NF-κB通路的变化探讨乳癌术后方对乳腺癌小鼠肺转移的影响[J].中华中医药学刊, 2016, 34(12):2864-2867.
张葳,陈君君,杨姣,等.通关藤注射液通过调控ATF3表达对紫杉醇抗乳腺癌细胞的增效作用[J].中成药, 2022, 44(6):1774-1782.
张小帅,杨柳青,王晓艳,等.通关藤注射液逆转乳腺癌多柔比星耐药的作用机制[J].中草药, 2024, 55(15):5112-5124.
Vyas D, Deshpande K, Chaturvedi L, et al. Rapid extensive recurrence of triple negative breast cancer:Are both therapy and cancer biology the culprit?[J]. J Clin Med Res,2016, 8(2):162-167.
Wu H, Wei G L, Luo L X, et al. Ginsenoside Rg3nanoparticles with permeation enhancing based chitosan derivatives were encapsulated with doxorubicin by thermosensitive hydrogel and anti-cancer evaluation of peritumoral hydrogel injection combined with PD-L1antibody[J]. Biomater Res, 2022, 26(1):77.
Wu L Q, Ding W S, Wang X P, et al. Interference KRT17reverses doxorubicin resistance in triple-negative breast cancer cells by Wnt/β-catenin signaling pathway[J].Genes Genomics, 2023, 45(10):1329-1338.
牛涛,周逢海.炎症与肿瘤微环境[J].中南大学学报:医学版, 2023, 48(12):1899-1913.
Wang S R, Wang Z Y, Li Z Y, et al. A catalytic immune activator based on magnetic nanoparticles to reprogram the immunoecology of breast cancer from“cold” to“hot” state[J]. Adv Healthc Mater, 2022, 11(21):2201240.
Pang Y H, Shi R Z, Chan L J, et al. The combination of the HDAC1 inhibitor SAHA and doxorubicin has synergic efficacy in triple negative breast cancer in vivo[J].Pharmacol Res, 2023, 196:106926.
Bai H T, Feng L, Schmid F. Macrophage-based cancer immunotherapy:Challenges and opportunities[J]. Exp Cell Res, 2024, 442(1):114198.
McWhorter R, Bonavida B. The role of TAMs in the regulation of tumor cell resistance to chemotherapy[J].Crit Rev Oncog, 2024, 29(4):97-125.
Heintzman D R, Fisher E L, Rathmell J C.Microenvironmental influences on T cell immunity in cancer and inflammation[J]. Cell Mol Immunol, 2022,19(3):316-326.
Miyazaki T, Suzuki G, Yamamura K I. The role of macrophages in antigen presentation and T cell tolerance[J]. Int Immunol, 1993, 5(9):1023-1033.
Rawlings J S, Rosler K M, Harrison D A. The JAK/STAT signaling pathway[J]. J Cell Sci, 2004, 117(8):1281-1283.
Xiong N T, Wu H M, Yu Z K. Advancements and challenges in triple-negative breast cancer:A comprehensive review of therapeutic and diagnostic strategies[J]. Front Oncol, 2024, 14:1405491.
Jin W. Role of JAK/STAT3 signaling in the regulation of metastasis, the transition of cancer stem cells, and chemoresistance of cancer by epithelial-mesenchymal transition[J]. Cells, 2020, 9(1):217.
Manore S G, Doheny D L, Wong G L, et al. IL-6/JAK/STAT3 signaling in breast cancer metastasis:Biology and treatment[J]. Front Oncol, 2022, 12:866014.
Jiang H, Pang J J, Li T Y, et al. PD-1 regulates the antitumor immune function of macrophages through JAK2-STAT3 signaling pathway in colorectal cancer tumor microenvironment[J]. J Transl Med, 2025, 23(1):502.
NíGabhann J, Hams E, Smith S, et al. Btk regulates macrophage polarization in response to lipopolysaccharide[J]. PLoS One, 2014, 9(1):e85834.
Martinet V, Tonon S, Torres D, et al. Type I interferons regulate eomesodermin expression and the development of unconventional memory CD8+T cells[J]. Nat Commun,2015(6):7089.
Meng X W, Cheng Z L, Lu Z Y, et al. MX2:Identification and systematic mechanistic analysis of a novel immunerelated biomarker for systemic lupus erythematosus[J].Front Immunol, 2022, 13:978851.
Le S T, Marusina A I, Merleev A A, et al. SPP1hi macrophages, NKG7 T cells, CCL5hi fibroblasts, and IgM plasma cells are dominant features of necrobiosis[J]. JCI Insight, 2025, 10(4):e178766.
Yang Y L, Zhang Y, Li D D, et al. RNF144A functions as a tumor suppressor in breast cancer through ubiquitin ligase activity-dependent regulation of stability and oncogenic functions of HSPA2[J]. Cell Death Differ,2020, 27(3):1105-1118.

2026年第57卷第9期

PDF下载

引用本文

BibTeX

文章信息

doi: 10.7501/j.issn.0253-2670.2026.09.014

接收时间：2026-01-05
首发时间：2026-09-09

补充材料

相关文章

文章信息

作者

出版历史

收稿日期：2026-01-05

基金

作者信息

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/zcy/CN/10.7501/j.issn.0253-2670.2026.09.014

分享至

全文二维码

扫描看全文

引用本文

BibTeX

本文的引用情况

2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

关闭全屏

BibTeX
EndNote
RefWorks
TxT