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Войти в исследования Zhen ci yan jiu = Acupuncture research · 2025
[Effects of heat-reinforcing acupuncture on Drp1/Mff pathway activity and mitochondrial quality in rabbits with cold-dampness rheumatoid arthritis].
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Jing Wei-Yao, Xu Cong-Hui, Liu Cui, Liu Li-Mei, Yuan Bo, Zhang Xing-Hua, Zhang Feng-Fan, Chen Ping, DU Xiao-Zheng
Аннотация
OBJECTIVES: To observe the efficacy of heat-reinforcing acupuncture at "Zusanli" (ST36) in rabbits with cold-dampness rheumatoid arthritis (RA) and its impact on mitochondrial quality and the dynamin-related protein 1 (Drp1)/mitochondrial fission factor (Mff) pathway in knee synovial tissue.
METHODS: Twenty-four New Zealand white rabbits were randomly divided into a normal group, a model group, a heat-reinforcing acupuncture group, and an inhibitor group, with 6 rabbits in each group. A cold-dampness RA rabbit model was established using complete Freund's adjuvant-ovalbumin mixture combined with an artificial climate chamber. Rabbits of the heat-reinforcing acupuncture group received heat-reinforcing acupuncture at ST36 once daily for 14 d. Rabbits of the inhibitor group was intraperitoneally injected with the mitochondrial division inhibitor Mdivi-1 (2.5 mg/kg) once every 2 d for a total of 7 times. Rabbits of the normal and model groups were handled and restrained in the same manner but received no other interventions. Knee arthritis scores, mechanical pain thresholds, and joint circumference were assessed after modeling and intervention. Hematoxylin-eosin (HE) staining was used to observe pathological changes in knee synovial tissue. Transmission electron microscopy was used to observe the ultrastructure of mitochondria in synovial cells. Luciferase method was used to measure ATP content in knee synovial tissue. Immunofluorescence staining was used to detect mitochondrial reactive oxygen species (mtROS) expression and Drp1/Mff co-localization in synovial tissue, and Western blot was used to measure the relative expression levels of phosphorylated(p)-Drp1 s616/Drp1 and Mff protein.
RESULTS: Compared with the normal group, the model group showed increased arthritis scores, joint circumference, mtROS expression, p-Drp1 s616/Drp1 and Mff protein expression, and Drp1/Mff co-localization in knee synovial tissue(P<0.01), while mechanical pain thresholds and ATP content in synovial tissue were significantly decreased (P<0.01). Following acupuncture intervention, compared with the model group, the heat-reinforcing acupuncture group and the inhibitor group showed significantly reduced arthritis scores, joint circumference, mtROS expression , p-Drp1 s616/Drp1 and Mff protein expression, and Drp1/Mff co-localization in synovial tissue(P<0.01), while mechanical pain thresholds and ATP content in synovial tissue were significantly increased (P<0.01). In the model group, synovial tissue showed significant synovial cell proliferation, with visible inflammatory cell infiltration and synovial matrix proliferation in the synovium;synovial cells had indistinct mitochondrial cristae, mitochondrial swelling/rupture, and mitochondrial fragmentation into small bodies. These changes in synovial tissue were alleviated in both the heat-reinforcing acupuncture group and the inhibitor group.
CONCLUSIONS: Heat-reinforcing acupuncture can significantly improve symptoms in RA rabbits with cold-dampness and enhance mitochondrial quality in synovial cells, which may be related to the inhibition of Drp1/Mff pathway activity.
METHODS: Twenty-four New Zealand white rabbits were randomly divided into a normal group, a model group, a heat-reinforcing acupuncture group, and an inhibitor group, with 6 rabbits in each group. A cold-dampness RA rabbit model was established using complete Freund's adjuvant-ovalbumin mixture combined with an artificial climate chamber. Rabbits of the heat-reinforcing acupuncture group received heat-reinforcing acupuncture at ST36 once daily for 14 d. Rabbits of the inhibitor group was intraperitoneally injected with the mitochondrial division inhibitor Mdivi-1 (2.5 mg/kg) once every 2 d for a total of 7 times. Rabbits of the normal and model groups were handled and restrained in the same manner but received no other interventions. Knee arthritis scores, mechanical pain thresholds, and joint circumference were assessed after modeling and intervention. Hematoxylin-eosin (HE) staining was used to observe pathological changes in knee synovial tissue. Transmission electron microscopy was used to observe the ultrastructure of mitochondria in synovial cells. Luciferase method was used to measure ATP content in knee synovial tissue. Immunofluorescence staining was used to detect mitochondrial reactive oxygen species (mtROS) expression and Drp1/Mff co-localization in synovial tissue, and Western blot was used to measure the relative expression levels of phosphorylated(p)-Drp1 s616/Drp1 and Mff protein.
RESULTS: Compared with the normal group, the model group showed increased arthritis scores, joint circumference, mtROS expression, p-Drp1 s616/Drp1 and Mff protein expression, and Drp1/Mff co-localization in knee synovial tissue(P<0.01), while mechanical pain thresholds and ATP content in synovial tissue were significantly decreased (P<0.01). Following acupuncture intervention, compared with the model group, the heat-reinforcing acupuncture group and the inhibitor group showed significantly reduced arthritis scores, joint circumference, mtROS expression , p-Drp1 s616/Drp1 and Mff protein expression, and Drp1/Mff co-localization in synovial tissue(P<0.01), while mechanical pain thresholds and ATP content in synovial tissue were significantly increased (P<0.01). In the model group, synovial tissue showed significant synovial cell proliferation, with visible inflammatory cell infiltration and synovial matrix proliferation in the synovium;synovial cells had indistinct mitochondrial cristae, mitochondrial swelling/rupture, and mitochondrial fragmentation into small bodies. These changes in synovial tissue were alleviated in both the heat-reinforcing acupuncture group and the inhibitor group.
CONCLUSIONS: Heat-reinforcing acupuncture can significantly improve symptoms in RA rabbits with cold-dampness and enhance mitochondrial quality in synovial cells, which may be related to the inhibition of Drp1/Mff pathway activity.