Journal of Traditional Chinese Medicine ›› 2026, Vol. 46 ›› Issue (4): 832-843.DOI: 10.19852/j.cnki.jtcm.2026.04.006
• Original Articles • Previous Articles Next Articles
Electroacupuncture enhances CD8+ T cells recruitment via modulating sympathetic nervous system to suppress tumor growth in mice bearing triple-negative breast cancer
TIAN Yehong1, ZHANG Qiaoli2, ZHANG Rui1, GAO Tao1, LI Nanzhi1, HUANG Jinchang2, JIANG Xin2(
), QIU Xiaowei3()
- 1
College of Acupuncture and Tuina ,Shaanxi University of Chinese Medicine Xianyang 712000, China
2Department of Acupuncture and Minimally Invasive Oncology ,The Third Affiliated Hospital of Beijing University of Chinese Medicine Beijing 100029, China
3Department of Encephalopathy Medicine ,BeiJing Chaoyang Intergrative Medicine Rescue and First Aid Hospital Beijing 100022, China
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Received:2025-06-16Accepted:2025-11-04Online:2026-08-15Published:2026-08-08 -
Contact:QIU Xiaowei, Department of Encephalopathy Medicine, Beijing Chaoyang Intergrative Medicine Rescue and First Aid Hospital, Beijing 100022, China. dcqiuxiaowei@sina.com;
JIANG Xin, Department of Acupuncture and Minimally Invasive Oncology, the Third Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100029, China. jxbucm@163.com,Telephone: +86-18182538201; +86-18311371983
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Supported by:National Natural Science Foundation of China: Exploring the Anti-tumor Mechanism of Peritumoral Needling Promoting CD8+ T Lymphocyte Recruitment of Breast Cancer Based on Neural Reprogramming(82305396);Scientific Research Project of Shaanxi Academy of Basic Research: Study on the Mechanism of Sympathetic Nerve Reprogramming Promoting CD8+ T Cell Recruitment in Triple-negative Breast Cancer Mice for Peritumoral Electroacupuncture Based on Histone Deacetylase 1 (HDAC1) Histone Modification(22JHQ089)
Cite this article
TIAN Yehong, ZHANG Qiaoli, ZHANG Rui, GAO Tao, LI Nanzhi, HUANG Jinchang, JIANG Xin, QIU Xiaowei. Electroacupuncture enhances CD8+ T cells recruitment via modulating sympathetic nervous system to suppress tumor growth in mice bearing triple-negative breast cancer[J]. Journal of Traditional Chinese Medicine, 2026, 46(4): 832-843.
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Figure 1 EA inhibited tumor growth and increased the recruitment of CD8+ T cells in mice with TNBC A: tumor growth curve; B: bioluminescence imaging; C: total fluorescence flux of the tumor burden; D: comparison of tumor weight; E: FCM analysis of CD8+ T cells in the blood; F: proportions of CD8+ T cells in the blood; G: FCM analysis of CD8+ T cells in the spleen; H: proportions of CD8+ T cells in the spleen; I: FCM analysis of CD8+ T cells in the tumor; J: proportions of CD8+ T cells in the tumor; K: representative IF images of CD8 (red) and CCL5 (green) in the tumor tissue (× 200); L: proportions of CD8+ T cells in the tumor by IF staining; M: relative expression of CCL5 in the tumor by IF staining; N: tumorous levels of the CCL5 by ELISA; O: serum CCL5 by ELISA; B1, E1, G1, I1, K1: model group; B2, E2, G2, I2, K2: EA group. EA group: underwent peritumoral EA with 2/30 Hz alternating frequencies under isoflurane anesthesia; Model group: only anesthetized with the same concentration of isoflurane. EA: electroacupuncture; TNBC: triple-negative breast cancer; 4T1-luc: luciferase-labeled 4T1 cell line; FCM: flow cytometry; IF: immunofluorescence; CCL5: C-C motif chemokine ligand 5; ELISA: enzyme-linked immunosorbent assay; DAPI: 4',6-diamidino-2-phenylindole. All quantitative data are expressed as mean ± standard error of the mean (n = 6); intergroup comparisons were performed based on Shapiro-Wilk normality and Levene’s homogeneity tests, with Student’s t-test for eligible data and Welch’s t-test or Wilcoxon rank-sum test otherwise. aP < 0.01 and bP < 0.001, compared with the model group.
Figure 1 EA inhibited tumor growth and increased the recruitment of CD8+ T cells in mice with TNBC A: tumor growth curve; B: bioluminescence imaging; C: total fluorescence flux of the tumor burden; D: comparison of tumor weight; E: FCM analysis of CD8+ T cells in the blood; F: proportions of CD8+ T cells in the blood; G: FCM analysis of CD8+ T cells in the spleen; H: proportions of CD8+ T cells in the spleen; I: FCM analysis of CD8+ T cells in the tumor; J: proportions of CD8+ T cells in the tumor; K: representative IF images of CD8 (red) and CCL5 (green) in the tumor tissue (× 200); L: proportions of CD8+ T cells in the tumor by IF staining; M: relative expression of CCL5 in the tumor by IF staining; N: tumorous levels of the CCL5 by ELISA; O: serum CCL5 by ELISA; B1, E1, G1, I1, K1: model group; B2, E2, G2, I2, K2: EA group. EA group: underwent peritumoral EA with 2/30 Hz alternating frequencies under isoflurane anesthesia; Model group: only anesthetized with the same concentration of isoflurane. EA: electroacupuncture; TNBC: triple-negative breast cancer; 4T1-luc: luciferase-labeled 4T1 cell line; FCM: flow cytometry; IF: immunofluorescence; CCL5: C-C motif chemokine ligand 5; ELISA: enzyme-linked immunosorbent assay; DAPI: 4',6-diamidino-2-phenylindole. All quantitative data are expressed as mean ± standard error of the mean (n = 6); intergroup comparisons were performed based on Shapiro-Wilk normality and Levene’s homogeneity tests, with Student’s t-test for eligible data and Welch’s t-test or Wilcoxon rank-sum test otherwise. aP < 0.01 and bP < 0.001, compared with the model group.
Figure 2 EA decreased intratumoral sympathetic innervation in tumor-bearing mice A: HE-stained images of peritumoral connective tissue (× 200); B: HE-stained images of tumor stroma (× 200); C: IF-stained images of TH (green) and DAPI (blue) in the tumor tissue (× 200); D: number of TH-marked sympathetic nerve; E: TH+ expression area per microscope field; F: representative IF images of β2-AR (red) and DAPI (blue) in the tumor tissue (× 200); G: relative expression of β2-AR in the mouse tumor by IF; H: tumorous levels of NE by ELISA; A1, B1, C1, F1: model group; A2, B2, C2, F2: EA group. EA group: underwent peritumoral EA with 2/30 Hz alternating frequencies under isoflurane anesthesia; Model group: only anesthetized with the same concentration of isoflurane. EA: electroacupuncture; HE: hematoxylin-eosin; TH: tyrosine hydrolase; DAPI: 4',6-diamidino-2-phenylindole; β2-AR: beta-2 adrenergic receptor; IF: immunofluorescence; NE: norepinephrine; ELISA: enzyme-linked immunosorbent assay. All quantitative data are presented as mean ± standard error of the mean (n = 6); intergroup comparisons were carried out based on Shapiro-Wilk normality and Levene’s homogeneity tests, with Student’s t-test for eligible data and Welch’s t-test or Wilcoxon rank-sum test otherwise. aP < 0.001 and bP < 0.01, compared with the model group.
Figure 2 EA decreased intratumoral sympathetic innervation in tumor-bearing mice A: HE-stained images of peritumoral connective tissue (× 200); B: HE-stained images of tumor stroma (× 200); C: IF-stained images of TH (green) and DAPI (blue) in the tumor tissue (× 200); D: number of TH-marked sympathetic nerve; E: TH+ expression area per microscope field; F: representative IF images of β2-AR (red) and DAPI (blue) in the tumor tissue (× 200); G: relative expression of β2-AR in the mouse tumor by IF; H: tumorous levels of NE by ELISA; A1, B1, C1, F1: model group; A2, B2, C2, F2: EA group. EA group: underwent peritumoral EA with 2/30 Hz alternating frequencies under isoflurane anesthesia; Model group: only anesthetized with the same concentration of isoflurane. EA: electroacupuncture; HE: hematoxylin-eosin; TH: tyrosine hydrolase; DAPI: 4',6-diamidino-2-phenylindole; β2-AR: beta-2 adrenergic receptor; IF: immunofluorescence; NE: norepinephrine; ELISA: enzyme-linked immunosorbent assay. All quantitative data are presented as mean ± standard error of the mean (n = 6); intergroup comparisons were carried out based on Shapiro-Wilk normality and Levene’s homogeneity tests, with Student’s t-test for eligible data and Welch’s t-test or Wilcoxon rank-sum test otherwise. aP < 0.001 and bP < 0.01, compared with the model group.
Figure 3 EA recruited CD8+ T cells to alleviate tumor progression from chronic stress-induced sympathetic activation, while sympathetic denervation abolished the antitumor effect A: tumor growth curve under exposure of CRS; B: bioluminescence imaging of the tumor; C: total fluorescence flux of the tumor; D: comparison of tumor weight; E: FCM analysis of tumorous CD8+ T cells; F: proportions of tumorous CD8+ T cells; G: IF images of CD8 (red) and CCL5 (green) in the tumor tissue (× 200); H: proportions of tumorous CD8+ T cells by IF; I: relative expression of tumorous CCL5 by IF; J: tumorous levels of the CCL5 by ELISA; K: serum CCL5 by ELISA; L: tumor growth curve following the sympathetic denervation; M: bioluminescence imaging; N: total fluorescence intensity of the tumor; O: comparison of tumor weight; B1, E1, G1: SE group; B2, E2, G2: CRS group; B3, E3, G3: CRS + EA group; M1: vehi group; M2: 6-OHDA group; M3: vehi + EA group; M4: 6-OHDA + EA group. SE group: raised in the standard environment; CRS group: underwent 2-h daily restraint stress from 1 week before 4T1-luc cell inoculation until experiment end; CRS + EA group: received peritumoral EA stimulation based on the CRS group; vehi group: received 0.2 mL saline with 0.01% ascorbate; 6-OHDA group: received 100 mg/kg 6-OHDA to ablate sympathetic innervation; vehi + EA group: received peritumoral EA based on the vehi group; 6-OHDA + EA: received peritumoral EA based on the 6-OHDA group. EA: electroacupuncture; CRS: chronic restraint stress; FCM: flow cytometry; IF: immunofluorescence; CCL5: C-C motif chemokine ligand 5; ELISA: enzyme-linked immunosorbent assay; 6-OHDA: 6-hydroxydopamine; Vehi: vehicle; SE: standard environment. All quantitative data are presented as mean ± standard error of the mean (n = 6); multiple groups were compared using one-way analysis of variance (ANOVA), followed by Welch's ANOVA, Tukey's honestly significant difference, or the Kruskal-Wallis test as needed; the Games-Howell test was used for post hoc analysis after ANOVA to assess group mean differences. aP < 0.05, bP < 0.01 and cP < 0.001, compared with the CRS + EA group; dP < 0.001, compared with the SE group; eP < 0.001, compared with the CRS group; fP < 0.01, compared with the SE group; gP < 0.05 and hP < 0.001, compared with the vehi group; iP < 0.001, compared with the vehi + EA group.
Figure 3 EA recruited CD8+ T cells to alleviate tumor progression from chronic stress-induced sympathetic activation, while sympathetic denervation abolished the antitumor effect A: tumor growth curve under exposure of CRS; B: bioluminescence imaging of the tumor; C: total fluorescence flux of the tumor; D: comparison of tumor weight; E: FCM analysis of tumorous CD8+ T cells; F: proportions of tumorous CD8+ T cells; G: IF images of CD8 (red) and CCL5 (green) in the tumor tissue (× 200); H: proportions of tumorous CD8+ T cells by IF; I: relative expression of tumorous CCL5 by IF; J: tumorous levels of the CCL5 by ELISA; K: serum CCL5 by ELISA; L: tumor growth curve following the sympathetic denervation; M: bioluminescence imaging; N: total fluorescence intensity of the tumor; O: comparison of tumor weight; B1, E1, G1: SE group; B2, E2, G2: CRS group; B3, E3, G3: CRS + EA group; M1: vehi group; M2: 6-OHDA group; M3: vehi + EA group; M4: 6-OHDA + EA group. SE group: raised in the standard environment; CRS group: underwent 2-h daily restraint stress from 1 week before 4T1-luc cell inoculation until experiment end; CRS + EA group: received peritumoral EA stimulation based on the CRS group; vehi group: received 0.2 mL saline with 0.01% ascorbate; 6-OHDA group: received 100 mg/kg 6-OHDA to ablate sympathetic innervation; vehi + EA group: received peritumoral EA based on the vehi group; 6-OHDA + EA: received peritumoral EA based on the 6-OHDA group. EA: electroacupuncture; CRS: chronic restraint stress; FCM: flow cytometry; IF: immunofluorescence; CCL5: C-C motif chemokine ligand 5; ELISA: enzyme-linked immunosorbent assay; 6-OHDA: 6-hydroxydopamine; Vehi: vehicle; SE: standard environment. All quantitative data are presented as mean ± standard error of the mean (n = 6); multiple groups were compared using one-way analysis of variance (ANOVA), followed by Welch's ANOVA, Tukey's honestly significant difference, or the Kruskal-Wallis test as needed; the Games-Howell test was used for post hoc analysis after ANOVA to assess group mean differences. aP < 0.05, bP < 0.01 and cP < 0.001, compared with the CRS + EA group; dP < 0.001, compared with the SE group; eP < 0.001, compared with the CRS group; fP < 0.01, compared with the SE group; gP < 0.05 and hP < 0.001, compared with the vehi group; iP < 0.001, compared with the vehi + EA group.
Figure 4 Chemical sympathetic ablation hindered CD8+ T cells recruitment of EA A: FCM analysis of CD8+ T cells in spleen; B: proportions of CD8+ T cells in the spleen; C: FCM analysis of CD8+ T cells in the tumor; D: proportions of CD8+ T cells in the tumor; E: representative IF images of CD8 (red), CCL5 (green) and DAPI (blue) in the tumor tissue (× 200); F: proportions of CD8+ T cells in the tumor of mice detected by IF staining; G: relative expression of CCL5 in the tumor of mice detected by IF; H: tumorous levels of the CCL5 detected by ELISA; I: serum CCL5 detected by ELISA; A1, C1, E1: vehi group; A2, C2, E2: 6-OHDA group; A3, C3, E3: vehi + EA group; A4, C4, E4: 6-OHDA+EA group. vehi group: received 0.2 mL saline with 0.01% ascorbate; 6-OHDA group: received 100 mg/kg 6-OHDA to ablate sympathetic innervation; vehi + EA group: received peritumoral EA based on the vehi group; 6-OHDA + EA: received peritumoral EA based on the 6-OHDA group. EA: electroacupuncture; FCM: flow cytometry; IF: immunofluorescence; CCL5: C-C motif chemokine ligand 5; DAPI: 4',6-diamidino-2-phenylindole; 6-OHDA: 6-hydroxydopamine; ELISA: enzyme-linked immunosorbent assay; Vehi: vehicle. All quantitative data are presented as mean ± standard error of the mean (n = 6); multiple groups were compared using one-way analysis of variance, followed by Tukey's honestly significant difference. aP < 0.05 and bP < 0.001, compared with the vehi group; cP < 0.001, compared with the vehi + EA group; dP < 0.001, compared with the 6-OHDA group; eP < 0.01, compared with the vehi group.
Figure 4 Chemical sympathetic ablation hindered CD8+ T cells recruitment of EA A: FCM analysis of CD8+ T cells in spleen; B: proportions of CD8+ T cells in the spleen; C: FCM analysis of CD8+ T cells in the tumor; D: proportions of CD8+ T cells in the tumor; E: representative IF images of CD8 (red), CCL5 (green) and DAPI (blue) in the tumor tissue (× 200); F: proportions of CD8+ T cells in the tumor of mice detected by IF staining; G: relative expression of CCL5 in the tumor of mice detected by IF; H: tumorous levels of the CCL5 detected by ELISA; I: serum CCL5 detected by ELISA; A1, C1, E1: vehi group; A2, C2, E2: 6-OHDA group; A3, C3, E3: vehi + EA group; A4, C4, E4: 6-OHDA+EA group. vehi group: received 0.2 mL saline with 0.01% ascorbate; 6-OHDA group: received 100 mg/kg 6-OHDA to ablate sympathetic innervation; vehi + EA group: received peritumoral EA based on the vehi group; 6-OHDA + EA: received peritumoral EA based on the 6-OHDA group. EA: electroacupuncture; FCM: flow cytometry; IF: immunofluorescence; CCL5: C-C motif chemokine ligand 5; DAPI: 4',6-diamidino-2-phenylindole; 6-OHDA: 6-hydroxydopamine; ELISA: enzyme-linked immunosorbent assay; Vehi: vehicle. All quantitative data are presented as mean ± standard error of the mean (n = 6); multiple groups were compared using one-way analysis of variance, followed by Tukey's honestly significant difference. aP < 0.05 and bP < 0.001, compared with the vehi group; cP < 0.001, compared with the vehi + EA group; dP < 0.001, compared with the 6-OHDA group; eP < 0.01, compared with the vehi group.
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