Objective:To observe the effects of moxibustion at Shenshu(BL23)and Zusanli(ST36)on Toll-like receptor 4(TLR4)-myeloid differentiation factor 88(Myd88)signaling pathway-mediated inflammatory factors in the synovial ti...Objective:To observe the effects of moxibustion at Shenshu(BL23)and Zusanli(ST36)on Toll-like receptor 4(TLR4)-myeloid differentiation factor 88(Myd88)signaling pathway-mediated inflammatory factors in the synovial tissue of ankle joints of rats with rheumatoid arthritis(RA),and to explore the molecular and biological mechanisms underlying the anti-inflammatory and analgesic effects.Methods:A total of 24 male Sprague-Dawley(SD)rats were randomly divided into a normal group,a model group,and a moxibustion group,with 8 rats in each group.The RA model was established with exposure to wind,cold,and damp environmental factors,along with Freund’s complete adjuvant.After three days of modeling,mild moxibustion was applied to bilateral Shenshu(BL23)and Zusanli(ST36)in the moxibustion group using moxa sticks of 0.9 cm in diameter for 30 min each time,once a day for 14 d.Structural changes in the synovial tissue and cells were then observed using hematoxylin-eosin staining and transmission electron microscopy,while immunohistochemistry analysis was used to detect tumor necrosis factor(TNF)-α,interleukin(IL)-17,IL-1β,and IL-6 levels.Moreover,the protein expression levels of Myd88,TLR4,and transient potential receptor vanilloid type 1(TRPV1)in the synovial tissue were detected using Western blotting,while their mRNA expression levels were detected using reverse transcription-polymerase chain reaction.Finally,the levels of IL-1β,IL-2,IL-6,IL-17A,and TNF-αin rat serum were detected using enzyme-linked immunosorbent assay.Results:Compared to the normal group,the model group exhibited notable pathological synovial tissue damage,along with significantly higher IL-1β,IL-6,and TNF-αlevels(P<0.01)and a slightly higher IL-17 content(P>0.05).Furthermore,the Myd88,TLR4,and TRPV1 protein and mRNA expression levels and serum IL-1β,IL-2,IL-6,IL-17A,and TNF-αlevels were all significantly higher in the model group than in the normal group(P<0.01).Compared to the model group,the moxibustion group exhibited a lower degree of synovial tissue pathological damage,along with significantly lower IL-1β,IL-6,and TNF-αlevels(P<0.05 or P<0.01)and a lower IL-17 content without statistical significance(P>0.05).Moreover,the Myd88,TLR4,and TRPV1 protein and mRNA expression levels,and serum IL-1β,IL-2,IL-6,IL-17A,and TNF-αlevels were all significantly lower in the moxibustion group than in the model group(P<0.01 or P<0.05).Conclusion:Mild moxibustion at Shenshu(BL23)and Zusanli(ST36)can effectively inhibit TLR4-MyD88 signaling pathway-mediated inflammatory factor expression in the synovial tissue of ankle joints of RA rats.Furthermore,the effect of moxibustion on synovial tissue inflammation in RA rats may be attributed to TRPV1 channel activation.展开更多
Objective:To investigate the protective effect of moxibustion on joints and its influence on the expression levels of S100 calcium binding protein A8(S100A8),S100 calcium binding protein A9(S100A9),serum amyloid A1(SA...Objective:To investigate the protective effect of moxibustion on joints and its influence on the expression levels of S100 calcium binding protein A8(S100A8),S100 calcium binding protein A9(S100A9),serum amyloid A1(SAA1),and related inflammatory factors in rats with adjuvant arthritis(AA).Methods:Forty Wistar rats were randomly divided into a normal group,a model group,a moxibustion group,and a medication group,with 10 rats in each group.Except for the normal group,AA models were established in the other three groups by exposing rats to wind-cold-dampness environmental conditions combined with complete Freund’s adjuvant.After successful modeling,the moxibustion group received moxibustion intervention,while the medication group was administered tripterygium glycosides tablets via oral gavage.The normal and model groups underwent similar handling and fixation without additional interventions.After 15 d of intervention,hematoxylin-eosin staining was used to assess pathological changes in the knee joint synovial membrane.Western blotting was performed to detect the protein expression of S100A8,S100A9,and SAA1 in the synovial tissue.Enzyme-linked immunosorbent assay was used to measure the serum levels of interferon(IFN)-γ,interleukin(IL)-6,and IL-23.Results:Compared to the normal group,the model group exhibited significantly increased protein expression of S100A8,S100A9,and SAA1 in the knee joint synovial tissue,as well as elevated serum levels of IFN-γ,IL-6,and IL-23(P<0.01).Histopathological analysis revealed marked synovial hyperplasia and extensive infiltration of inflammatory cells in the model group.Compared to the model group,both the moxibustion and medication groups showed significant reductions in the protein expression of S100A8,S100A9,and SAA1 in the synovial tissue,as well as decreased serum levels of IFN-γ,IL-6,and IL-23(P<0.01).Additionally,synovial tissue in these two groups displayed minimal hyperplasia and only mild inflammatory cell infiltration.Notably,compared to the moxibustion group,the medication group exhibited significantly higher protein expression of S100A9 in the synovial tissue(P<0.05),while no significant differences were observed in the expression of S100A8,SAA1,or serum levels of IFN-γ,IL-6,and IL-23(P>0.05).Both intervention groups showed comparable degrees of synovial inflammation,clear tissue structure,and no obvious hyperplasia.Conclusion:Moxibustion can alleviate joint swelling and reduce inflammatory responses in AA rats.Its mechanism may involve regulating the protein expression of S100A8,S100A9,and SAA1 in the knee joint synovial tissue.展开更多
Objective:To observe the effect of moxibustion on the expression of N-methyl-D-aspartic acid(NMDA)receptor subtype 2B(NR2B)in the hippocampus of rheumatoid arthritis(RA)rats,and to explore the analgesic mechanisms of ...Objective:To observe the effect of moxibustion on the expression of N-methyl-D-aspartic acid(NMDA)receptor subtype 2B(NR2B)in the hippocampus of rheumatoid arthritis(RA)rats,and to explore the analgesic mechanisms of moxibustion in RA treatment.Methods:Sixty male Sprague-Dawley rats were randomly divided into a normal group,a model group,a moxibustion group,a moxibustion+NMDA receptor antagonist(AP-5)group,and a moxibustion+NMDA receptor agonist(NMDA)group,with 12 rats in each group.Except for the normal group,rats in the other four groups were treated with complete Freund's adjuvant in a windy,cold,and damp environment to replicate RA models.Rats in the moxibustion group received suspended moxibustion with moxa sticks at Shenshu(BL23)and Zusanli(ST36),and the two points were used alternately.After intraperitoneal injection of AP-5 or NMDA,rats in the moxibustion+AP-5 group and the moxibustion+NMDA group received the same moxibustion intervention as in the moxibustion group,once a day for 15 d.The thermal withdrawal latency(TWL)of rats in each group was detected before and after modeling and after the 15-day intervention.After the 15-day intervention,hematoxylin-eosin staining was performed to observe the pathological changes in knee joints.The real-time fluorescence quantitative polymerase chain reaction method was used to detect the mRNA expression of NR2B in the hippocampus;Western blotting assay was used to detect the protein and the phosphorylated protein expression of hippocampal NR2B.Results:The synovial tissue was proliferated,the synovial lining was significantly thickened,the pannus was formed,and the cartilage and bone tissues were significantly damaged in the model group.After intervention,the pathological morphology of the knee joints in the moxibustion group,the moxibustion+AP-5 group,and the moxibustion+NMDA group was significantly improved,and the improvement in the moxibustion+AP-5 group was more notable than that in the moxibustion+NMDA group.Compared with the normal group,the TWL was significantly decreased(P<0.01),and the mRNA,protein,and phosphorylated protein expression levels of hippocampal NR2B were significantly increased in the model group(P<0.01).Compared with the model group,the TWL of each intervention group was significantly increased(P<0.01 or P<0.05),and the mRNA,protein,and phosphorylated protein expression levels of hippocampal NR2B were significantly decreased(P<0.01).Compared with the moxibustion group,the TWL was significantly increased(P<0.01),and the mRNA,protein,and phosphorylated protein expression levels of hippocampal NR2B were significantly decreased in the moxibustion+AP-5 group(P<0.01);the TWL was significantly decreased(P<0.01),and the mRNA,protein,and phosphorylated protein expression levels of hippocampal NR2B were significantly increased in the moxibustion+NMDA group(P<0.01).Conclusion:Moxibustion reduces hyperalgesia in RA inflammatory rats.The analgesic effect may be related to the decrease in the expression and phosphorylation levels of NR2B in the hippocampus.展开更多
文摘Objective:To observe the effects of moxibustion at Shenshu(BL23)and Zusanli(ST36)on Toll-like receptor 4(TLR4)-myeloid differentiation factor 88(Myd88)signaling pathway-mediated inflammatory factors in the synovial tissue of ankle joints of rats with rheumatoid arthritis(RA),and to explore the molecular and biological mechanisms underlying the anti-inflammatory and analgesic effects.Methods:A total of 24 male Sprague-Dawley(SD)rats were randomly divided into a normal group,a model group,and a moxibustion group,with 8 rats in each group.The RA model was established with exposure to wind,cold,and damp environmental factors,along with Freund’s complete adjuvant.After three days of modeling,mild moxibustion was applied to bilateral Shenshu(BL23)and Zusanli(ST36)in the moxibustion group using moxa sticks of 0.9 cm in diameter for 30 min each time,once a day for 14 d.Structural changes in the synovial tissue and cells were then observed using hematoxylin-eosin staining and transmission electron microscopy,while immunohistochemistry analysis was used to detect tumor necrosis factor(TNF)-α,interleukin(IL)-17,IL-1β,and IL-6 levels.Moreover,the protein expression levels of Myd88,TLR4,and transient potential receptor vanilloid type 1(TRPV1)in the synovial tissue were detected using Western blotting,while their mRNA expression levels were detected using reverse transcription-polymerase chain reaction.Finally,the levels of IL-1β,IL-2,IL-6,IL-17A,and TNF-αin rat serum were detected using enzyme-linked immunosorbent assay.Results:Compared to the normal group,the model group exhibited notable pathological synovial tissue damage,along with significantly higher IL-1β,IL-6,and TNF-αlevels(P<0.01)and a slightly higher IL-17 content(P>0.05).Furthermore,the Myd88,TLR4,and TRPV1 protein and mRNA expression levels and serum IL-1β,IL-2,IL-6,IL-17A,and TNF-αlevels were all significantly higher in the model group than in the normal group(P<0.01).Compared to the model group,the moxibustion group exhibited a lower degree of synovial tissue pathological damage,along with significantly lower IL-1β,IL-6,and TNF-αlevels(P<0.05 or P<0.01)and a lower IL-17 content without statistical significance(P>0.05).Moreover,the Myd88,TLR4,and TRPV1 protein and mRNA expression levels,and serum IL-1β,IL-2,IL-6,IL-17A,and TNF-αlevels were all significantly lower in the moxibustion group than in the model group(P<0.01 or P<0.05).Conclusion:Mild moxibustion at Shenshu(BL23)and Zusanli(ST36)can effectively inhibit TLR4-MyD88 signaling pathway-mediated inflammatory factor expression in the synovial tissue of ankle joints of RA rats.Furthermore,the effect of moxibustion on synovial tissue inflammation in RA rats may be attributed to TRPV1 channel activation.
文摘Objective:To investigate the protective effect of moxibustion on joints and its influence on the expression levels of S100 calcium binding protein A8(S100A8),S100 calcium binding protein A9(S100A9),serum amyloid A1(SAA1),and related inflammatory factors in rats with adjuvant arthritis(AA).Methods:Forty Wistar rats were randomly divided into a normal group,a model group,a moxibustion group,and a medication group,with 10 rats in each group.Except for the normal group,AA models were established in the other three groups by exposing rats to wind-cold-dampness environmental conditions combined with complete Freund’s adjuvant.After successful modeling,the moxibustion group received moxibustion intervention,while the medication group was administered tripterygium glycosides tablets via oral gavage.The normal and model groups underwent similar handling and fixation without additional interventions.After 15 d of intervention,hematoxylin-eosin staining was used to assess pathological changes in the knee joint synovial membrane.Western blotting was performed to detect the protein expression of S100A8,S100A9,and SAA1 in the synovial tissue.Enzyme-linked immunosorbent assay was used to measure the serum levels of interferon(IFN)-γ,interleukin(IL)-6,and IL-23.Results:Compared to the normal group,the model group exhibited significantly increased protein expression of S100A8,S100A9,and SAA1 in the knee joint synovial tissue,as well as elevated serum levels of IFN-γ,IL-6,and IL-23(P<0.01).Histopathological analysis revealed marked synovial hyperplasia and extensive infiltration of inflammatory cells in the model group.Compared to the model group,both the moxibustion and medication groups showed significant reductions in the protein expression of S100A8,S100A9,and SAA1 in the synovial tissue,as well as decreased serum levels of IFN-γ,IL-6,and IL-23(P<0.01).Additionally,synovial tissue in these two groups displayed minimal hyperplasia and only mild inflammatory cell infiltration.Notably,compared to the moxibustion group,the medication group exhibited significantly higher protein expression of S100A9 in the synovial tissue(P<0.05),while no significant differences were observed in the expression of S100A8,SAA1,or serum levels of IFN-γ,IL-6,and IL-23(P>0.05).Both intervention groups showed comparable degrees of synovial inflammation,clear tissue structure,and no obvious hyperplasia.Conclusion:Moxibustion can alleviate joint swelling and reduce inflammatory responses in AA rats.Its mechanism may involve regulating the protein expression of S100A8,S100A9,and SAA1 in the knee joint synovial tissue.
文摘Objective:To observe the effect of moxibustion on the expression of N-methyl-D-aspartic acid(NMDA)receptor subtype 2B(NR2B)in the hippocampus of rheumatoid arthritis(RA)rats,and to explore the analgesic mechanisms of moxibustion in RA treatment.Methods:Sixty male Sprague-Dawley rats were randomly divided into a normal group,a model group,a moxibustion group,a moxibustion+NMDA receptor antagonist(AP-5)group,and a moxibustion+NMDA receptor agonist(NMDA)group,with 12 rats in each group.Except for the normal group,rats in the other four groups were treated with complete Freund's adjuvant in a windy,cold,and damp environment to replicate RA models.Rats in the moxibustion group received suspended moxibustion with moxa sticks at Shenshu(BL23)and Zusanli(ST36),and the two points were used alternately.After intraperitoneal injection of AP-5 or NMDA,rats in the moxibustion+AP-5 group and the moxibustion+NMDA group received the same moxibustion intervention as in the moxibustion group,once a day for 15 d.The thermal withdrawal latency(TWL)of rats in each group was detected before and after modeling and after the 15-day intervention.After the 15-day intervention,hematoxylin-eosin staining was performed to observe the pathological changes in knee joints.The real-time fluorescence quantitative polymerase chain reaction method was used to detect the mRNA expression of NR2B in the hippocampus;Western blotting assay was used to detect the protein and the phosphorylated protein expression of hippocampal NR2B.Results:The synovial tissue was proliferated,the synovial lining was significantly thickened,the pannus was formed,and the cartilage and bone tissues were significantly damaged in the model group.After intervention,the pathological morphology of the knee joints in the moxibustion group,the moxibustion+AP-5 group,and the moxibustion+NMDA group was significantly improved,and the improvement in the moxibustion+AP-5 group was more notable than that in the moxibustion+NMDA group.Compared with the normal group,the TWL was significantly decreased(P<0.01),and the mRNA,protein,and phosphorylated protein expression levels of hippocampal NR2B were significantly increased in the model group(P<0.01).Compared with the model group,the TWL of each intervention group was significantly increased(P<0.01 or P<0.05),and the mRNA,protein,and phosphorylated protein expression levels of hippocampal NR2B were significantly decreased(P<0.01).Compared with the moxibustion group,the TWL was significantly increased(P<0.01),and the mRNA,protein,and phosphorylated protein expression levels of hippocampal NR2B were significantly decreased in the moxibustion+AP-5 group(P<0.01);the TWL was significantly decreased(P<0.01),and the mRNA,protein,and phosphorylated protein expression levels of hippocampal NR2B were significantly increased in the moxibustion+NMDA group(P<0.01).Conclusion:Moxibustion reduces hyperalgesia in RA inflammatory rats.The analgesic effect may be related to the decrease in the expression and phosphorylation levels of NR2B in the hippocampus.
