Journal of Traditional Chinese Medicine ›› 2026, Vol. 46 ›› Issue (4): 844-853.DOI: 10.19852/j.cnki.jtcm.20251201.001
• Original Articles • Previous Articles Next Articles
Regulatory effects and mechanisms of the acupuncture-induced complement component 1q on the high mobility group box 1 -receptor for advanced glycation end products-nuclear factor kappa-B pathway at Zusanli (ST36)
KANG Xiaochun1, WU Sinuo1, LI Ningcen2, XU Wenyue1, XU Zhifang2, Thong Yikchih1, LI Peiyun1, TU Shiwei1, MA Ning3, LI Yanwei2, DOU Baomin1, TAN Wenjun4, GUO Jing5, GUO Yi2, PAN Xingfang2(
), LÜ Zhongxi2()
- 1
School of Acupuncture & Moxibustion and Tuina ,Tianjin University of Traditional Chinese Medicine Tianjin 301617, China
2Research Center of Experimental Acupuncture Science ,Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China; School of Acupuncture & Moxibustion and Tuina, Tianjin University of Traditional Chinese Medicine Tianjin 301617, China
3Institute of Traditional Chinese Medicine ,Suzuka University of Medical Science Suzuka 5100226, Japan
4Key Laboratory of Intelligent Computing in Medical Image ,Ministry of Education, Northeastern University Shenyang 110189, China
5School of Automation ,Guangdong University of technology Guangzhou 510006, China
-
Received:2025-05-10Accepted:2025-09-30Online:2026-08-15Published:2025-12-01 -
Contact:Associate professor LÜ Zhongxi, full professor PAN Xingfang, School of Acupuncture & Moxibustion and Tuina, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China. 1025117807@qq.com;panxingfang@163.com -
About author:KANG Xiaochun and WU Sinuo are co-first authors and contributed equally to this work -
Supported by:National Key R&D Program of China: Efficacy and Safety Verification of Intelligent Traditional Chinese Medicine Acupuncture Robot System(2022YFB4703100);National Natural Science Foundation of China: Study on the Effect and Mechanism of C1q Protein in the Ordered and Controlled Inflammatory Response in Acupuncture Area(82105023);Mechanism of electroacupuncture-driven Surface-modified Graded Cell-derived Glycosylated Exo-miR-146a targeting T Cell Immunoglobulin and Mucin Domain-containing Protein 3 to Regulate Exhausted T Cells and Alleviate Sepsis Immunosuppression(82305370);Initial Dynamic Regulation Mechanism of Acupuncture Effect via Physicochemical Coupling Network in Acupoint Microenvironment(82030125);Young Elite Scientists Sponsorship Program by China Association of Chinese Medicine: Initiation Mechanism of Acupuncture Analgesia Mediated by High Mobility Group Box 1 at Acupoints(2023-QNRC2-B04)
Cite this article
KANG Xiaochun, WU Sinuo, LI Ningcen, XU Wenyue, XU Zhifang, Thong Yikchih, LI Peiyun, TU Shiwei, MA Ning, LI Yanwei, DOU Baomin, TAN Wenjun, GUO Jing, GUO Yi, PAN Xingfang, LÜ Zhongxi. Regulatory effects and mechanisms of the acupuncture-induced complement component 1q on the high mobility group box 1 -receptor for advanced glycation end products-nuclear factor kappa-B pathway at Zusanli (ST36)[J]. Journal of Traditional Chinese Medicine, 2026, 46(4): 844-853.
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Figure 1 Effect of acupuncture on HMGB1, IL-10, and C1q proteins at Zusanli (ST36) A: concentration of HMGB1 proteins; B: concentration of IL-10 proteins; C: concentration of C1q proteins; A1, B1, C1: D1 after acupuncture; A2, B2, C2: D3 after acupuncture; A3, B3, C3: D5 after acupuncture; A4, B4, C4: D7 after acupuncture; D-F: the temporal changes of HMGB1, IL-10 and C1q proteins after acupuncture (D3 vs D1, D5 vs D3, D7 vs D5). D: HMGB1 proteins; E: IL-10 proteins; F: C1q proteins. WT group: wild-type mice without acupuncture intervention, ACU group: wild-type mice receiving bilateral Zusanli (ST36) acupuncture stimulation. Acupuncture was performed at Zusanli (ST36) using sterilized stainless-steel filiform needles in 8-min daily sessions for 1, 3, 5, or 7 d. HMGB1: high mobility group box 1; IL-10: interleukin-10; C1q: complement component 1q; D: day. Data are presented as mean ± standard deviation (n = 5-7 per group). The t-test was used for data analysis between two groups. aP < 0.05, bP < 0.01, cP < 0.001 versus the WT group.
Figure 1 Effect of acupuncture on HMGB1, IL-10, and C1q proteins at Zusanli (ST36) A: concentration of HMGB1 proteins; B: concentration of IL-10 proteins; C: concentration of C1q proteins; A1, B1, C1: D1 after acupuncture; A2, B2, C2: D3 after acupuncture; A3, B3, C3: D5 after acupuncture; A4, B4, C4: D7 after acupuncture; D-F: the temporal changes of HMGB1, IL-10 and C1q proteins after acupuncture (D3 vs D1, D5 vs D3, D7 vs D5). D: HMGB1 proteins; E: IL-10 proteins; F: C1q proteins. WT group: wild-type mice without acupuncture intervention, ACU group: wild-type mice receiving bilateral Zusanli (ST36) acupuncture stimulation. Acupuncture was performed at Zusanli (ST36) using sterilized stainless-steel filiform needles in 8-min daily sessions for 1, 3, 5, or 7 d. HMGB1: high mobility group box 1; IL-10: interleukin-10; C1q: complement component 1q; D: day. Data are presented as mean ± standard deviation (n = 5-7 per group). The t-test was used for data analysis between two groups. aP < 0.05, bP < 0.01, cP < 0.001 versus the WT group.
Figure 2 Acupuncture-induced C1q involved in regulating the inflammatory responses at Zusanli (ST36) A: representative hematoxylin-eosin staining images of Zusanli (ST36) tissues (Nuclei: blue, cytoplasm: red, Scale bar = 50 µm, n = 3, magnification × 40); A1-A3: the epidermis, dermis layers of the WT, ACU, and C1q-/-+ACU groups; A1:WT groups; A2: ACU groups; A3: C1q-/-+ACU groups; A4-A6: the muscle layers of the WT, ACU, and C1q-/-+ACU groups; A4: WT groups; A5: ACU groups; A6: C1q-/-+ACU groups; B: single-label immunofluorescence of p-p65 (p-p65: green, 1:100, DAPI: blue, Scale bar = 10 µm, n = 3, magnification × 40). B1-B3: representative merged images showing p-p65 with DAPI; B1: WT groups; B2: ACU groups; B3: C1q-/-+ACU groups; C: quantification of p-p65-positive cells by immunofluorescence (n = 3); D: protein concentration of HMGB1 at Zusanli (ST36) measured by ELISA (n = 5-6); E: protein concentration of IL-10 at Zusanli (ST36) measured by ELISA (n = 5-6). WT group: wild-type mice without acupuncture intervention, ACU group: wild-type mice receiving bilateral Zusanli (ST36) acupuncture stimulation, C1q-/-+ACU group: C1q knockout mice receiving acupuncture. Acupuncture was performed at Zusanli (ST36) using sterilized stainless-steel filiform needles in 8-min daily sessions for 7 d. HMGB1: high mobility group box 1; IL-10: interleukin-10; ELISA: enzyme-linked immunosorbent assay; p-p65: phosphorylated p65. Data are presented as mean ± standard deviation. One-way analysis of variance was used for multiple groups comparisons. If the data followed a normal distribution, the least significant difference test was used; otherwise, Dunnett’s T3 test was used. aP < 0.05, cP < 0.01 versus the WT group; bP < 0.05 versus the ACU group.
Figure 2 Acupuncture-induced C1q involved in regulating the inflammatory responses at Zusanli (ST36) A: representative hematoxylin-eosin staining images of Zusanli (ST36) tissues (Nuclei: blue, cytoplasm: red, Scale bar = 50 µm, n = 3, magnification × 40); A1-A3: the epidermis, dermis layers of the WT, ACU, and C1q-/-+ACU groups; A1:WT groups; A2: ACU groups; A3: C1q-/-+ACU groups; A4-A6: the muscle layers of the WT, ACU, and C1q-/-+ACU groups; A4: WT groups; A5: ACU groups; A6: C1q-/-+ACU groups; B: single-label immunofluorescence of p-p65 (p-p65: green, 1:100, DAPI: blue, Scale bar = 10 µm, n = 3, magnification × 40). B1-B3: representative merged images showing p-p65 with DAPI; B1: WT groups; B2: ACU groups; B3: C1q-/-+ACU groups; C: quantification of p-p65-positive cells by immunofluorescence (n = 3); D: protein concentration of HMGB1 at Zusanli (ST36) measured by ELISA (n = 5-6); E: protein concentration of IL-10 at Zusanli (ST36) measured by ELISA (n = 5-6). WT group: wild-type mice without acupuncture intervention, ACU group: wild-type mice receiving bilateral Zusanli (ST36) acupuncture stimulation, C1q-/-+ACU group: C1q knockout mice receiving acupuncture. Acupuncture was performed at Zusanli (ST36) using sterilized stainless-steel filiform needles in 8-min daily sessions for 7 d. HMGB1: high mobility group box 1; IL-10: interleukin-10; ELISA: enzyme-linked immunosorbent assay; p-p65: phosphorylated p65. Data are presented as mean ± standard deviation. One-way analysis of variance was used for multiple groups comparisons. If the data followed a normal distribution, the least significant difference test was used; otherwise, Dunnett’s T3 test was used. aP < 0.05, cP < 0.01 versus the WT group; bP < 0.05 versus the ACU group.
Figure 3 Mechanism of C1q regulation of the inflammatory response at Zusanli (ST36) A: representative Western blot bands showing protein expression of RAGE, LAIR-1, and SHP-1 at Zusanli (ST36) (n = 5-6); B: quantification of LAIR-1 normalized to GAPDH; C: quantification of RAGE normalized to GAPDH; D: quantification of SHP-1normalized to GAPDH; E: double-label immunofluorescence of M1 macrophages (M0: F4/80, red, 1:50; M1: CD86, green, 1:100; Merged: F4/80, CD86, and DAPI; Scale bar = 100 µm; magnification ×40, n = 3); E1: F4/80-label of WT groups; E2: F4/80-label of ACU groups; E3: F4/80-label of C1q-/-+ACU groups, E4: CD86-label of WT groups; E5: CD86-label of ACU groups; E6: CD86-label of C1q-/-+ACU groups; E7: merged of WT groups; E8: merged of ACU groups; E9: merged of C1q-/-+ACU groups, F: double-label immunofluorescence of M2 macrophages (M0: F4/80, red, 1:50; M2: CD206, green, 1:50, Merged: F4/80, CD206, and DAPI; Scale bar = 100 µm; magnification ×40, n = 3); F1: F4/80-label of WT groups; F2: F4/80-label of ACU groups; F3: F4/80-label of C1q-/-+ACU groups, F4: CD206-label of WT groups; F5: CD206-label of ACU groups; F6: CD206-label of C1q-/-+ACU groups; F7: merged of WT groups; F8: merged of ACU groups; F9: merged of C1q-/-+ACU groups; G : quantitative immunofluorescence analysis of M1 macrophages at Zusanli (ST36) (n = 3); H: quantitative immunofluorescence analysis of M2 macrophages at Zusanli (ST36) (n = 3). WT group: wild-type mice without acupuncture intervention, ACU group: wild-type mice receiving bilateral Zusanli (ST36) acupuncture stimulation, C1q-/-+ACU group: C1q knockout mice receiving acupuncture. Acupuncture was performed at Zusanli (ST36) using sterilized stainless-steel filiform needles in 8-min daily sessions for 7 d. RAGE: the receptor for advanced glycation end-products; LAIR-1: leukocyte-associated immunoglobulin-like receptor 1; SHP-1: src homology 2 domain-containing protein tyrosine phosphatase 1; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; M0: Naïve Macrophage; M1: classical activated macrophages; M2: alternatively activated macrophages. Data are presented as mean ± standard deviation. One-way analysis of variance was used for multiple groups comparisons. If the data followed a normal distribution, the least significant difference test was used; otherwise, Dunnett’s T3 test was used. cP < 0.01, aP < 0.001 versus the WT group; bP < 0.05, dP < 0.001 versus the ACU group.
Figure 3 Mechanism of C1q regulation of the inflammatory response at Zusanli (ST36) A: representative Western blot bands showing protein expression of RAGE, LAIR-1, and SHP-1 at Zusanli (ST36) (n = 5-6); B: quantification of LAIR-1 normalized to GAPDH; C: quantification of RAGE normalized to GAPDH; D: quantification of SHP-1normalized to GAPDH; E: double-label immunofluorescence of M1 macrophages (M0: F4/80, red, 1:50; M1: CD86, green, 1:100; Merged: F4/80, CD86, and DAPI; Scale bar = 100 µm; magnification ×40, n = 3); E1: F4/80-label of WT groups; E2: F4/80-label of ACU groups; E3: F4/80-label of C1q-/-+ACU groups, E4: CD86-label of WT groups; E5: CD86-label of ACU groups; E6: CD86-label of C1q-/-+ACU groups; E7: merged of WT groups; E8: merged of ACU groups; E9: merged of C1q-/-+ACU groups, F: double-label immunofluorescence of M2 macrophages (M0: F4/80, red, 1:50; M2: CD206, green, 1:50, Merged: F4/80, CD206, and DAPI; Scale bar = 100 µm; magnification ×40, n = 3); F1: F4/80-label of WT groups; F2: F4/80-label of ACU groups; F3: F4/80-label of C1q-/-+ACU groups, F4: CD206-label of WT groups; F5: CD206-label of ACU groups; F6: CD206-label of C1q-/-+ACU groups; F7: merged of WT groups; F8: merged of ACU groups; F9: merged of C1q-/-+ACU groups; G : quantitative immunofluorescence analysis of M1 macrophages at Zusanli (ST36) (n = 3); H: quantitative immunofluorescence analysis of M2 macrophages at Zusanli (ST36) (n = 3). WT group: wild-type mice without acupuncture intervention, ACU group: wild-type mice receiving bilateral Zusanli (ST36) acupuncture stimulation, C1q-/-+ACU group: C1q knockout mice receiving acupuncture. Acupuncture was performed at Zusanli (ST36) using sterilized stainless-steel filiform needles in 8-min daily sessions for 7 d. RAGE: the receptor for advanced glycation end-products; LAIR-1: leukocyte-associated immunoglobulin-like receptor 1; SHP-1: src homology 2 domain-containing protein tyrosine phosphatase 1; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; M0: Naïve Macrophage; M1: classical activated macrophages; M2: alternatively activated macrophages. Data are presented as mean ± standard deviation. One-way analysis of variance was used for multiple groups comparisons. If the data followed a normal distribution, the least significant difference test was used; otherwise, Dunnett’s T3 test was used. cP < 0.01, aP < 0.001 versus the WT group; bP < 0.05, dP < 0.001 versus the ACU group.
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