OBJECTIVE:to investigate the anti-arthritic effects of lappaconitine(LA)on adjuvant-induced arthritis in Sprague-Dawley rats and its possible involvement in the regulation of M1/M2 macrophage balance through the P2X7 ...OBJECTIVE:to investigate the anti-arthritic effects of lappaconitine(LA)on adjuvant-induced arthritis in Sprague-Dawley rats and its possible involvement in the regulation of M1/M2 macrophage balance through the P2X7 receptor(P2X7r).METHODS:Rats were immunized with complete Freund's adjuvant and then intraperitoneally administered LA(2,4,or 8 mg·kg^(-1)·d^(-1))or methotrexate(0.5 mg/kg per 3 d)for 14 d.The anti-arthritic effects of LA were evaluated through arthritis index(AI)assessment,ankle diameter measurement,and histopathological staining analysis.The analgesic effect of LA on arthritis was measured using mechanical withdrawal threshold testing and gait scoring.The impacts of LA on macrophage polarization,the expression of pro-/anti-inflammatory cytokines and P2X7r were analyzed using quantitative real-time polymerase chain reaction,enzyme-linked immunosorbent assay,and Western blotting.RESULTS:LA treatment significantly reduced AI scores,paw swelling,joint destruction,and inflammatory cell infiltration,and alleviated arthritis pain.Additionally,LA promoted a balanced M1/M2 ratio by increasing the m RNA expression level of M2 marker arginase 1 and decreasing those of M1 markers inducible nitric oxide synthase and interleukin(IL)-1βin synovial tissues.Furthermore,LA lowered the levels of three M1-related cytokines,namely tumor necrosis factor-α,IL-1βand IL-18,and raised the level of the M2-related cytokine IL-10.Further research showed that treatment with LA inhibited the expression of P2X7r.CONCLUSION:Our findings indicate that the notable therapeutic and analgesic effects of LA on AIA rats are exerted through balancing the M1/M2 ratio,probably via P2X7r.展开更多
Objective:To investigate the mechanism by which moxibustion regulates the expression of inflammatory cytokines in ulcerative colitis(UC)rats through the P2X7 receptor(P2X7R)/nuclear factor-kappa B(NF-κB)pathway.Metho...Objective:To investigate the mechanism by which moxibustion regulates the expression of inflammatory cytokines in ulcerative colitis(UC)rats through the P2X7 receptor(P2X7R)/nuclear factor-kappa B(NF-κB)pathway.Methods:UC was induced using dextran sulfate sodium(DSS)in both wild-type(WT)and P2X7R knockout(KO)mice.General health conditions,pathological changes,and periodic acid-Schiff(PAS)staining of the colonic tissues were analyzed.Immunohistochemistry was used to detect NF-κB p65 protein expression in colonic tissues.Male Sprague-Dawley(SD)rats were randomly assigned to four groups:normal,model,normal+herb-partitioned moxibustion,and model+herb-partitioned moxibustion.UC was induced in rats by cyclic DSS administration.Rats in the herb-partitioned moxibustion group received moxibustion at the bilateral Tianshu(ST25)and Qihai(RN6)acupoints.The effects of herb-partitioned moxibustion were evaluated regarding general health conditions and histopathological alterations in colon tissue.The protein expression of P2X7R and NF-κB p65 in colonic tissues was determined by immunohistochemistry,whereas interleukin(IL)-10 mRNA levels were quantified using real-time quantitative polymerase chain reaction(RT-qPCR).Furthermore,enzyme-linked immunosorbent assay(ELISA)was used to measure serum concentrations of tumor necrosis factor-alpha(TNF-α)and IL-6.Results:Colonic epithelial damage and inflammatory cell infiltration were significantly reduced in P2X7R KO mice compared to WT mice,along with reduced expression of NF-κB p65 protein in colonic tissues(P<0.05).Moxibustion improves histopathological damage,goblet cell number,and intestinal mucus secretion in rats with UC.Compared to the normal group,the model group exhibited increased histopathological scores,serum TNF-α,and IL-6 levels,as well as elevated P2X7R and NF-κB p65 protein expression in colonic tissues(P<0.05).In comparison to the model group,the model+herb-partitioned moxibustion group demonstrated significantly lower histopathological scores,reduced serum TNF-αand IL-6 levels,and decreased P2X7R and NF-κB p65 protein expression(P<0.05).Conclusions:Moxibustion at“Tianshu”and“Qihai”acupoints may inhibit the levels of IL-6 and TNF-αinflammatory factors and reduce inflammation in the UC colonic mucosa by regulating the P2X7R/NF-κB p65 pathway in UC colonic tissues.展开更多
文摘OBJECTIVE:to investigate the anti-arthritic effects of lappaconitine(LA)on adjuvant-induced arthritis in Sprague-Dawley rats and its possible involvement in the regulation of M1/M2 macrophage balance through the P2X7 receptor(P2X7r).METHODS:Rats were immunized with complete Freund's adjuvant and then intraperitoneally administered LA(2,4,or 8 mg·kg^(-1)·d^(-1))or methotrexate(0.5 mg/kg per 3 d)for 14 d.The anti-arthritic effects of LA were evaluated through arthritis index(AI)assessment,ankle diameter measurement,and histopathological staining analysis.The analgesic effect of LA on arthritis was measured using mechanical withdrawal threshold testing and gait scoring.The impacts of LA on macrophage polarization,the expression of pro-/anti-inflammatory cytokines and P2X7r were analyzed using quantitative real-time polymerase chain reaction,enzyme-linked immunosorbent assay,and Western blotting.RESULTS:LA treatment significantly reduced AI scores,paw swelling,joint destruction,and inflammatory cell infiltration,and alleviated arthritis pain.Additionally,LA promoted a balanced M1/M2 ratio by increasing the m RNA expression level of M2 marker arginase 1 and decreasing those of M1 markers inducible nitric oxide synthase and interleukin(IL)-1βin synovial tissues.Furthermore,LA lowered the levels of three M1-related cytokines,namely tumor necrosis factor-α,IL-1βand IL-18,and raised the level of the M2-related cytokine IL-10.Further research showed that treatment with LA inhibited the expression of P2X7r.CONCLUSION:Our findings indicate that the notable therapeutic and analgesic effects of LA on AIA rats are exerted through balancing the M1/M2 ratio,probably via P2X7r.
基金funded by the National Natural Science Foundation of China(82174501,82105012,82205293,82205262)Shanghai Municipal Natural Science Foundation(22ZR1458400)+2 种基金Shanghai Talent Development Fund Project(2021058)Shanghai University of Traditional Chinese Medicine Science and Technology Development Project(23KFL111)State Administration of Traditional Chinese Medicine high-level key discipline construction project(zyyzdxk-2023068)。
文摘Objective:To investigate the mechanism by which moxibustion regulates the expression of inflammatory cytokines in ulcerative colitis(UC)rats through the P2X7 receptor(P2X7R)/nuclear factor-kappa B(NF-κB)pathway.Methods:UC was induced using dextran sulfate sodium(DSS)in both wild-type(WT)and P2X7R knockout(KO)mice.General health conditions,pathological changes,and periodic acid-Schiff(PAS)staining of the colonic tissues were analyzed.Immunohistochemistry was used to detect NF-κB p65 protein expression in colonic tissues.Male Sprague-Dawley(SD)rats were randomly assigned to four groups:normal,model,normal+herb-partitioned moxibustion,and model+herb-partitioned moxibustion.UC was induced in rats by cyclic DSS administration.Rats in the herb-partitioned moxibustion group received moxibustion at the bilateral Tianshu(ST25)and Qihai(RN6)acupoints.The effects of herb-partitioned moxibustion were evaluated regarding general health conditions and histopathological alterations in colon tissue.The protein expression of P2X7R and NF-κB p65 in colonic tissues was determined by immunohistochemistry,whereas interleukin(IL)-10 mRNA levels were quantified using real-time quantitative polymerase chain reaction(RT-qPCR).Furthermore,enzyme-linked immunosorbent assay(ELISA)was used to measure serum concentrations of tumor necrosis factor-alpha(TNF-α)and IL-6.Results:Colonic epithelial damage and inflammatory cell infiltration were significantly reduced in P2X7R KO mice compared to WT mice,along with reduced expression of NF-κB p65 protein in colonic tissues(P<0.05).Moxibustion improves histopathological damage,goblet cell number,and intestinal mucus secretion in rats with UC.Compared to the normal group,the model group exhibited increased histopathological scores,serum TNF-α,and IL-6 levels,as well as elevated P2X7R and NF-κB p65 protein expression in colonic tissues(P<0.05).In comparison to the model group,the model+herb-partitioned moxibustion group demonstrated significantly lower histopathological scores,reduced serum TNF-αand IL-6 levels,and decreased P2X7R and NF-κB p65 protein expression(P<0.05).Conclusions:Moxibustion at“Tianshu”and“Qihai”acupoints may inhibit the levels of IL-6 and TNF-αinflammatory factors and reduce inflammation in the UC colonic mucosa by regulating the P2X7R/NF-κB p65 pathway in UC colonic tissues.
