Journal of traditional Chinese medicine = Chung i tsa chih ying wen pan · 2026
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.
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Feng Hao, Jiajia Sang, Chuanyu Peng, Jun H U, Zijian W U, Ling H U, Yihuang G U
Аннотация
OBJECTIVE: To investigate the effects of moxibustion at Zusanli (ST36) on ferroptosis-related proteins in synovial inflammation of rats with rheumatoid arthritis (RA) and verify the mechanism by cell experiments.
METHODS: A RA rat model was established using a "disease-syndrome integration" modeling method. The rats were randomly divided into a blank control group (Control group) and a model replication group. Rats with successfully replicated RA models were further divided into a model control group (RA group) and a moxibustion treatment group (RA + Mox group). After intervention, the rats' ankle swelling degree and small animal ultrasound examination were conducted and synovial tissue was examined for pathology, lipid reactive oxygen species (ROS) detection, ferrous ion Fe2+ double staining, and immunohistochemistry analysis. Serum from three groups of rats were extracted and used for cellular assays, cell counting kit-8 and Western blot validation.
RESULTS: (a) Compared to Control group rats, RA rats showed significantly increased footpad swelling (P < 0.01) and joint inflammation levels (P < 0.001); compared to RA group, the RA + Mox rats showed significantly decreased footpad swelling (P < 0.05) and joint inflammation levels (P < 0.01). (b) There was no difference in lipid ROS levels in the synovial tissue of RA rats compared to Control group rats (P > 0.05), but the lipid ROS levels in the synovial tissue of RA + Mox rats were significantly increased (P < 0.01). Compared to Control group rats, RA rats had significantly lower Fe2+ levels (P < 0.05); compared to RA rats, the Fe2+ levels in RA+Mox rats were significantly increased (P < 0.01). There was no significant difference in beclin 1(BECN1) expression in the synovial tissue of RA rats compared to Control group rats (P >0.05), but BECN1 expression in the synovial tissue of RA + Mox rats was significantly increased (P < 0.01) compared to RA rats. Compared to Control group rats, RA rats had significantly higher Solute Carrier Family 7 Member 11 Gene (SLC7A11) levels in synovial tissue (P < 0.05); compared to RA rats, the SLC7A11 levels in RA + Mox rats were significantly decreased (P < 0.05). (c) In the lipopolysaccharides (10 ng/mL) model, 50% concentration of RA rats serum promoted human RA fibroblast-like synovial cell line (MH7A) proliferation (48 h) (P < 0.05) and 50% concentration of RA + Mox rat serum inhibited MH7A proliferation (48, 72 h) (P < 0.001). Both 25% and 50% concentrations of RA + Mox rat serum upregulated BECN1 and downregulated SLC7A11 expression in MH7A.
CONCLUSION: Moxibustion at Zusanli (ST36) suggests a potential to relieve footpad swelling and synovial inflammation in RA rats, alleviate synovial tissue hyperplasia and thickening. It may induce lipid ROS accumulation in synovial tissue, upregulating Fe2+ levels, upregulating BECN1 expression, and downregulating SLC7A11 expression in RA rats. Moreover, 50% concentration of rat serum after moxibustion inhibited the proliferation of synovial fibroblasts and regulated the expression of ferroptosis-related proteins.
METHODS: A RA rat model was established using a "disease-syndrome integration" modeling method. The rats were randomly divided into a blank control group (Control group) and a model replication group. Rats with successfully replicated RA models were further divided into a model control group (RA group) and a moxibustion treatment group (RA + Mox group). After intervention, the rats' ankle swelling degree and small animal ultrasound examination were conducted and synovial tissue was examined for pathology, lipid reactive oxygen species (ROS) detection, ferrous ion Fe2+ double staining, and immunohistochemistry analysis. Serum from three groups of rats were extracted and used for cellular assays, cell counting kit-8 and Western blot validation.
RESULTS: (a) Compared to Control group rats, RA rats showed significantly increased footpad swelling (P < 0.01) and joint inflammation levels (P < 0.001); compared to RA group, the RA + Mox rats showed significantly decreased footpad swelling (P < 0.05) and joint inflammation levels (P < 0.01). (b) There was no difference in lipid ROS levels in the synovial tissue of RA rats compared to Control group rats (P > 0.05), but the lipid ROS levels in the synovial tissue of RA + Mox rats were significantly increased (P < 0.01). Compared to Control group rats, RA rats had significantly lower Fe2+ levels (P < 0.05); compared to RA rats, the Fe2+ levels in RA+Mox rats were significantly increased (P < 0.01). There was no significant difference in beclin 1(BECN1) expression in the synovial tissue of RA rats compared to Control group rats (P >0.05), but BECN1 expression in the synovial tissue of RA + Mox rats was significantly increased (P < 0.01) compared to RA rats. Compared to Control group rats, RA rats had significantly higher Solute Carrier Family 7 Member 11 Gene (SLC7A11) levels in synovial tissue (P < 0.05); compared to RA rats, the SLC7A11 levels in RA + Mox rats were significantly decreased (P < 0.05). (c) In the lipopolysaccharides (10 ng/mL) model, 50% concentration of RA rats serum promoted human RA fibroblast-like synovial cell line (MH7A) proliferation (48 h) (P < 0.05) and 50% concentration of RA + Mox rat serum inhibited MH7A proliferation (48, 72 h) (P < 0.001). Both 25% and 50% concentrations of RA + Mox rat serum upregulated BECN1 and downregulated SLC7A11 expression in MH7A.
CONCLUSION: Moxibustion at Zusanli (ST36) suggests a potential to relieve footpad swelling and synovial inflammation in RA rats, alleviate synovial tissue hyperplasia and thickening. It may induce lipid ROS accumulation in synovial tissue, upregulating Fe2+ levels, upregulating BECN1 expression, and downregulating SLC7A11 expression in RA rats. Moreover, 50% concentration of rat serum after moxibustion inhibited the proliferation of synovial fibroblasts and regulated the expression of ferroptosis-related proteins.