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
Microglia are involved in the inflammatory response and retinal ganglion cell damage in glaucoma.Here,we investigated how microglia proliferate and migrate in a mouse model of chronic ocular hypertension(COH).In COH r...Microglia are involved in the inflammatory response and retinal ganglion cell damage in glaucoma.Here,we investigated how microglia proliferate and migrate in a mouse model of chronic ocular hypertension(COH).In COH retinas,the microglial proliferation that occurred was inhibited by the P2X7 receptor(P2X7R)blocker BBG or P2X7R knockout,but not by the P2X4R blocker 5-BDBD.Treatment of primary cultured microglia with BzATP,a P2X7R agonist,mimicked the effects of cell proliferation and migration in COH retinas through the intracellular MEK/ERK signaling pathway.Transwell migration assays showed that the P2X4R agonist CTP induced microglial migration,which was completely blocked by 5-BDBD.In vivo and in vitro experiments demonstrated that ATP,released from activated Müller cells through connexin43 hemichannels,acted on P2X7R to induce microglial proliferation,and acted on P2X4R/P2X7R(mainly P2X4R)to induce microglial migration.Our results suggest that inhibiting the interaction of Müller cells and microglia may attenuate microglial proliferation and migration in glaucoma.展开更多
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.
基金This work was supported by grants from the National Natural Science Foundation of China(81790642 and 31872765)the Shanghai Municipal Science and Technology Major Project(2018SHZDZX01)ZJ Lab,and the Shanghai Center for Brain Science and Brain-Inspired Technology.
文摘Microglia are involved in the inflammatory response and retinal ganglion cell damage in glaucoma.Here,we investigated how microglia proliferate and migrate in a mouse model of chronic ocular hypertension(COH).In COH retinas,the microglial proliferation that occurred was inhibited by the P2X7 receptor(P2X7R)blocker BBG or P2X7R knockout,but not by the P2X4R blocker 5-BDBD.Treatment of primary cultured microglia with BzATP,a P2X7R agonist,mimicked the effects of cell proliferation and migration in COH retinas through the intracellular MEK/ERK signaling pathway.Transwell migration assays showed that the P2X4R agonist CTP induced microglial migration,which was completely blocked by 5-BDBD.In vivo and in vitro experiments demonstrated that ATP,released from activated Müller cells through connexin43 hemichannels,acted on P2X7R to induce microglial proliferation,and acted on P2X4R/P2X7R(mainly P2X4R)to induce microglial migration.Our results suggest that inhibiting the interaction of Müller cells and microglia may attenuate microglial proliferation and migration in glaucoma.
