Journal of Traditional Chinese Medicine ›› 2026, Vol. 46 ›› Issue (3): 594-604.DOI: 10.19852/j.cnki.jtcm.20260306.001
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Effect of moxibustion at Zusanli (ST36) on beclin 1-solute carrier family 7 member 11 gene ferroptosis-related proteins in synovial inflammation of rats with rheumatoid arthritis
HAO Feng1, SANG Jiajia2, PENG Chuanyu3, HU Jun1, WU Zijian4, HU Ling4, GU Yihuang1(
)
- 1
School of Acupuncture-Moxibustion and Tuina ,Nanjing University of Chinese Medicine Nanjing 210023, China
2Department of Tuina ,the Affiliated Hospital of Nanjing University of Traditional Chinese Medicine/Jiangsu Provincial Hospital of Traditional Chinese Medicine Nanjing 210029, China
3School of Acupuncture-Moxibustion and Tuina ,Anhui University of Chinese Medicine Hefei 230038, China
4Institute of Acupuncture and Meridian ,Anhui Academy of Traditional Chinese Medicine Hefei 230038, China
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Received:2025-04-04Accepted:2025-09-05Online:2026-06-15Published:2026-03-06 -
Contact:Dr. GU Yihuang, School of Acupuncture-Moxibustion and Tuina, Nanjing University of Chinese Medicine, Nanjing 210023, China. gyh5196411@163.com, Telephone: +86-25-85811663 -
Supported by:Natural Science Foundation-funded Project: Autophagy-dependent Ferroptosis of Fibroblast-like Synoviocytes Induced by Recombinant beclin 1-Solute Carrier Family 7 Member 11 Gene in the Mechanism of Moxibustion in Treating Rheumatoid Arthritis(82274645)
Cite this article
HAO Feng, SANG Jiajia, PENG Chuanyu, HU Jun, WU Zijian, HU Ling, GU Yihuang. Effect of moxibustion at Zusanli (ST36) on beclin 1-solute carrier family 7 member 11 gene ferroptosis-related proteins in synovial inflammation of rats with rheumatoid arthritis[J]. Journal of Traditional Chinese Medicine, 2026, 46(3): 594-604.
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Figure 1 Body weight and paw condition of rats, synovial tissue condition of rats and HE staining of synovial tissue in rats of different groups A: body weight during the experiment; B: measurement of joint swelling degree (B1) and arthritis index (B2); C: ultrasonic imaging of synovial tissue; D: HE staining of synovial tissue (× 200, scale bar = 50 μm). C1, D1: Control group; C2, D2: RA group; C3, D3: RA + Mox group. Control group: healthy rats, no intervention; RA group: RA model rats, RA model rats were established using a "disease-syndrome integration" modeling method, specifically by exposing the rats to "wind, cold, and dampness" environmental factors combined with 0.15 mL of complete Freund's adjuvant; RA + Mox group: RA rats with moxibustion treatment, moxibustion applied at bilateral Zusanli (ST36) for 20 min/d, once a day, for a continuous 15 d of treatment. HE: hematoxylin-eosin; Control: blank control group; RA: rheumatoid arthritis; Mox: moxibustion. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with RA group, aP < 0.05, dP < 0.01, fP< 0.001; compared with Control group, bP < 0.05, cP < 0.01, eP< 0.001.
Figure 1 Body weight and paw condition of rats, synovial tissue condition of rats and HE staining of synovial tissue in rats of different groups A: body weight during the experiment; B: measurement of joint swelling degree (B1) and arthritis index (B2); C: ultrasonic imaging of synovial tissue; D: HE staining of synovial tissue (× 200, scale bar = 50 μm). C1, D1: Control group; C2, D2: RA group; C3, D3: RA + Mox group. Control group: healthy rats, no intervention; RA group: RA model rats, RA model rats were established using a "disease-syndrome integration" modeling method, specifically by exposing the rats to "wind, cold, and dampness" environmental factors combined with 0.15 mL of complete Freund's adjuvant; RA + Mox group: RA rats with moxibustion treatment, moxibustion applied at bilateral Zusanli (ST36) for 20 min/d, once a day, for a continuous 15 d of treatment. HE: hematoxylin-eosin; Control: blank control group; RA: rheumatoid arthritis; Mox: moxibustion. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with RA group, aP < 0.05, dP < 0.01, fP< 0.001; compared with Control group, bP < 0.05, cP < 0.01, eP< 0.001.
Figure 2 Lipid ROS and Fe2+ levels in synovial tissue of rats in different groups A: lipid ROS in synovial tissue were detected by immunofluorescence (× 400, scale bar = 20 μm); B: Fe2+ levels in synovial tissue were detected by immunofluorescence (× 400, scale bar = 20 μm); A1, B1: Control group; A2, B2: RA group; A3, B3: RA + Mox group; A4: the mean density value of ROS in synovial tissue; B4: the mean density value of Fe2+ in synovial tissue. Control group: healthy rats, no intervention; RA group: RA model rats, RA model rats were established using a "disease-syndrome integration" modeling method, specifically by exposing the rats to "wind, cold, and dampness" environmental factors combined with 0.15 mL of complete Freund's adjuvant; RA + Mox group: RA rats with moxibustion treatment, moxibustion applied at bilateral Zusanli (ST36) for 20 min/d, once a day, for a continuous 15 d of treatment. Control: blank control group; RA: rheumatoid arthritis; Mox: moxibustion; ROS: lipid reactive oxygen species. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with Control group, aP < 0.05; compared with RA group, bP < 0.05.
Figure 2 Lipid ROS and Fe2+ levels in synovial tissue of rats in different groups A: lipid ROS in synovial tissue were detected by immunofluorescence (× 400, scale bar = 20 μm); B: Fe2+ levels in synovial tissue were detected by immunofluorescence (× 400, scale bar = 20 μm); A1, B1: Control group; A2, B2: RA group; A3, B3: RA + Mox group; A4: the mean density value of ROS in synovial tissue; B4: the mean density value of Fe2+ in synovial tissue. Control group: healthy rats, no intervention; RA group: RA model rats, RA model rats were established using a "disease-syndrome integration" modeling method, specifically by exposing the rats to "wind, cold, and dampness" environmental factors combined with 0.15 mL of complete Freund's adjuvant; RA + Mox group: RA rats with moxibustion treatment, moxibustion applied at bilateral Zusanli (ST36) for 20 min/d, once a day, for a continuous 15 d of treatment. Control: blank control group; RA: rheumatoid arthritis; Mox: moxibustion; ROS: lipid reactive oxygen species. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with Control group, aP < 0.05; compared with RA group, bP < 0.05.
Figure 3 Expression of BECN1 and SLC7A11 in synovial tissue of rats in different groups A: expression of BECN1 in synovial tissue was detected by IHC (× 200, scale bar = 50 μm); B: expression of SLC7A11 in synovial tissue was detected by IHC (× 200, scale bar = 50 μm); A1, B1: Control group; A2, B2: RA group; A3, B3: RA + Mox group; A4: the mean density value of BECN1 in synovial tissue; B4: the mean density value of SLC7A11 in synovial tissue. Control group: healthy rats, no intervention; RA group: RA model rats, RA model rats were established using a "disease-syndrome integration" modeling method, specifically by exposing the rats to "wind, cold, and dampness" environmental factors combined with 0.15 mL of complete Freund's adjuvant; RA + Mox group: RA rats with moxibustion treatment, moxibustion applied at bilateral Zusanli (ST36) for 20 min/d, once a day, for a continuous 15 d of treatment. Control: blank control group; RA: rheumatoid arthritis; Mox: moxibustion; BECN1: beclin 1; SLC7A11: solute carrier family 7 member 11 gene; IHC: immunohistochemical. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with Control group, aP < 0.05; compared with RA group, bP < 0.05.
Figure 3 Expression of BECN1 and SLC7A11 in synovial tissue of rats in different groups A: expression of BECN1 in synovial tissue was detected by IHC (× 200, scale bar = 50 μm); B: expression of SLC7A11 in synovial tissue was detected by IHC (× 200, scale bar = 50 μm); A1, B1: Control group; A2, B2: RA group; A3, B3: RA + Mox group; A4: the mean density value of BECN1 in synovial tissue; B4: the mean density value of SLC7A11 in synovial tissue. Control group: healthy rats, no intervention; RA group: RA model rats, RA model rats were established using a "disease-syndrome integration" modeling method, specifically by exposing the rats to "wind, cold, and dampness" environmental factors combined with 0.15 mL of complete Freund's adjuvant; RA + Mox group: RA rats with moxibustion treatment, moxibustion applied at bilateral Zusanli (ST36) for 20 min/d, once a day, for a continuous 15 d of treatment. Control: blank control group; RA: rheumatoid arthritis; Mox: moxibustion; BECN1: beclin 1; SLC7A11: solute carrier family 7 member 11 gene; IHC: immunohistochemical. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with Control group, aP < 0.05; compared with RA group, bP < 0.05.
Figure 4 Protein levels of BECN1 and SLC7A11 in all groups of MH7A cell A. protein images of BECN1 and SLC7A11 in all groups of MH7A cell; B: quantitative analysis of BECN1 and SLC7A11 protein levels in different time points. B1: quantitative analysis of BECN1 at 48 h; B2: quantitative analysis of BECN1 at 72 h; B3: quantitative analysis of SLC7A11 at 48 h; B4: quantitative analysis of SLC7A11 at 72 h. Control group: basal medium; LPS group: MH7A with LPS stimulus (10 ng/mL, 100 μL per well); LPS + 25% RA + Mox group: MH7A with LPS stimulus (10 ng/mL, 100 μL per well) and 25% serum concentration (from RA rats treated with moxibustion); LPS + 50% RA + Mox group: MH7A with LPS stimulus (10 ng/mL, 100 μL per well) and 50% serum concentration (from RA rats treated with moxibustion). Control: blank control group; RA: rheumatoid arthritis; Mox: moxibustion; BECN1: beclin 1; SLC7A11: solute carrier family 7 member 11 gene; LPS: lipopolysaccharides. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 5, replicate wells per group). Compared with Control group, aP < 0.05; compared with LPS group, bP < 0.01; compared with LPS + 25% RA + Mox group, cP < 0.01.
Figure 4 Protein levels of BECN1 and SLC7A11 in all groups of MH7A cell A. protein images of BECN1 and SLC7A11 in all groups of MH7A cell; B: quantitative analysis of BECN1 and SLC7A11 protein levels in different time points. B1: quantitative analysis of BECN1 at 48 h; B2: quantitative analysis of BECN1 at 72 h; B3: quantitative analysis of SLC7A11 at 48 h; B4: quantitative analysis of SLC7A11 at 72 h. Control group: basal medium; LPS group: MH7A with LPS stimulus (10 ng/mL, 100 μL per well); LPS + 25% RA + Mox group: MH7A with LPS stimulus (10 ng/mL, 100 μL per well) and 25% serum concentration (from RA rats treated with moxibustion); LPS + 50% RA + Mox group: MH7A with LPS stimulus (10 ng/mL, 100 μL per well) and 50% serum concentration (from RA rats treated with moxibustion). Control: blank control group; RA: rheumatoid arthritis; Mox: moxibustion; BECN1: beclin 1; SLC7A11: solute carrier family 7 member 11 gene; LPS: lipopolysaccharides. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 5, replicate wells per group). Compared with Control group, aP < 0.05; compared with LPS group, bP < 0.01; compared with LPS + 25% RA + Mox group, cP < 0.01.
| 1. |
Patel D, Shelbaya A, Cheung R, Aggarwal J, Park SH, Coindreau J. Cost-effectiveness of early treatment with originator biologics or their biosimilars after methotrexate failure in patients with established rheumatoid arthritis. Adv Ther 2019; 36: 2086-95.
DOI PMID |
| 2. |
Smolen JS, Aletaha D, McInnes IB. Rheumatoid arthritis. Lancet 2016; 388: 2023-38.
DOI PMID |
| 3. |
Tang JL, Liu BY, Ma KW. Traditional Chinese Medicine. Lancet 2008; 372: 1938-40.
DOI URL |
| 4. |
Zhang P, Li J, Han Y, Yu XW, Qin L. Traditional Chinese Medicine in the treatment of rheumatoid arthritis: a general review. Rheumatol Int 2010; 30: 713-8.
DOI PMID |
| 5. | Wang X, Wu P, Luo Y, et al. Moxibustion for rheumatoid arthritis and its effect on related negative emotions. Zhong Guo Zhen Jiu 2022; 42: 1221-5. |
| 6. | Tao SY, Tang J, Wang X, Jiang NN, Li Y, Wu P. Moxibustion as an adjunctive treatment for rheumatoid arthritis and its effects on the serum levels of SOST and β-catenin. Zhong Guo Zhen Jiu 2023; 43: 1384-9. |
| 7. |
Hao F, Wang Q, Liu L, et al. Effect of moxibustion on autophagy and the inflammatory response of synovial cells in rheumatoid arthritis model rat. J Tradit Chin Med 2022; 42: 73-82.
PMID |
| 8. | Hao F, Wu LB, Hu J, et al. Effect of moxibustion on PI3K/Akt/mTOR signaling pathway in foot-pad synovium in rats with rheumatoid arthritis. Zhong Guo Zhen Jiu 2020; 40: 1211-6. |
| 9. | Hao F, Liu L, Sang JJ, et al. Effect of moxibustion on expression of autophagy related molecules in synovial tissues of rheumatoid arthritis rats. Zhen Ci Yan Jiu 2020; 45: 793-8. |
| 10. |
Wang T, Peng C, Gao D, Zhang C, Hao F, He L. Moxibustion's impact on ferroptosis regulation: a key to relieving inflammatory injury in rheumatoid arthritis. Rejuvenation Res 2025; 28: 217-25.
DOI PMID |
| 11. |
Zhao T, Yang Q, Xi Y, et al. Ferroptosis in rheumatoid arthritis: a potential therapeutic strategy. Front Immunol 2022; 13: 779585.
DOI URL |
| 12. |
Xiang J, Chen H, Lin Z, Chen J, Luo L. Identification and experimental validation of ferroptosis-related gene SLC2A3 is involved in rheumatoid arthritis. Eur J Pharmacol 2023; 943: 175568.
DOI URL |
| 13. | Wang H, Zhang M, Hu Y, et al. Deciphering the role of ferroptosis in rheumatoid arthritis: synovial transcriptome analysis and immune infiltration correlation. Heliyon 2024; 10: e33648. |
| 14. | Shang J, Fan W, Dou Z, Wu L, Lu B, Qian J. The efficacy and safety of warming acupuncture and moxibustion on rheumatoid arthritis: a protocol for a systematic review and Meta-analysis. Medicine (Baltimore) 2020; 99: e21857. |
| 15. |
Chen X, Kang R, Kroemer G, Tang D. Organelle-specific regulation of ferroptosis. Cell Death Differ 2021; 28: 2843-56.
DOI PMID |
| 16. |
Koppula P, Zhuang L, Gan B. Cystine transporter SLC7A11/xCT in cancer: ferroptosis, nutrient dependency, and cancer therapy. Protein Cell 2021; 12: 599-620.
DOI |
| 17. |
Wu J, Feng Z, Chen L, et al. TNF antagonist sensitizes synovial fibroblasts to ferroptotic cell death in collagen-induced arthritis mouse models. Nat Commun 2022; 13: 676.
DOI PMID |
| 18. |
Han Z, Gao X, Wang Y, et al. Ultrasmall iron-quercetin metal natural product nanocomplex with antioxidant and macrophage regulation in rheumatoid arthritis. Acta Pharm Sin B 2023; 13: 1726-39.
DOI PMID |
| 19. |
Lee ES, Sul JH, Shin JM, et al. Reactive oxygen species-responsive dendritic cell-derived exosomes for rheumatoid arthritis. Acta Biomater 2021; 128: 462-73.
DOI URL |
| 20. |
Li Y, Liang Q, Zhou L, et al. An ROS-responsive artesunate prodrug nanosystem co-delivers dexamethasone for rheumatoid arthritis treatment through the HIF-1α/NF-κB cascade regulation of ROS scavenging and macrophage repolarization. Acta Biomater 2022; 152: 406-24.
DOI PMID |
| 21. |
Mizushima N, Levine B. Autophagy in human diseases. N Engl J Med 2020; 383: 1564-76.
DOI URL |
| 22. |
Matsuzawa-Ishimoto Y, Hwang S, Cadwell K. Autophagy and inflammation. Annu Rev Immunol 2018; 36: 73-101.
DOI PMID |
| 23. |
Liu J, Kuang F, Kroemer G, Klionsky DJ, Kang R, Tang D. Autophagy-dependent ferroptosis: machinery and regulation. Cell Chem Biol 2020; 27: 420-35.
DOI PMID |
| 24. |
Zhang PQ, Feng Q, Huang WY, Ou S. Protective effect of lappaconitine on Freund's complete adjuvant-induced arthritis exerted through P2X7 receptor-mediated regulation of M1/M2 balance in rats. J Tradit Chin Med 2025; 45: 39-48.
DOI |
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