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
The nod-like receptor family pyrin domain containing 3(NLRP3)inflammasome plays a crucial role in the prognosis of subarachnoid hemorrhage(SAH).WNK1 kinase negatively regulates NLRP3 in various inflammatory conditions...The nod-like receptor family pyrin domain containing 3(NLRP3)inflammasome plays a crucial role in the prognosis of subarachnoid hemorrhage(SAH).WNK1 kinase negatively regulates NLRP3 in various inflammatory conditions,but its role in early brain injury(EBI)after SAH remains unclear.In this study,we used an in vivo SAH model in rats/mice and AAV-WNK1 intraventricular injection to investigate its neuroprotective mechanisms.WNK1 expression was significantly reduced in SAH patient blood and SAH model brain tissue,correlating negatively with microglial activation.AAV-WNK1 alleviated brain edema,neuronal necrosis,behavioral deficits,and inflammation by inhibiting NLRP3 inflammasome activation.In hemin-stimulated BV-2 cells,WNK1 overexpression reduced NLRP3 activation and inflammatory cytokines.Chloride counteracted WNK1’s inhibitory effects,and WNK1 suppressed P2X7R-induced NLRP3 activation.Mechanistically,WNK1 functioned via the OXSR1/STK39 pathway.These findings highlight WNK1 as a key regulator of intracellular chloride balance and neuroinflammation,presenting a potential therapeutic target for SAH treatment.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supported by Grants from the Basic Research Program of Jiangsu(BK20240492)Lianyungang Science and Technology Plan Project(JCYJ2304)+2 种基金Lianyungang Aging Health Research Project(L202301)Doctoral Research Start-up Fund of the First Peopel’Hospital of Lianyungang City(BS202314)The funders had no role in study design,data collection and analysis,decision to publish,or preparation of the manuscript.
文摘The nod-like receptor family pyrin domain containing 3(NLRP3)inflammasome plays a crucial role in the prognosis of subarachnoid hemorrhage(SAH).WNK1 kinase negatively regulates NLRP3 in various inflammatory conditions,but its role in early brain injury(EBI)after SAH remains unclear.In this study,we used an in vivo SAH model in rats/mice and AAV-WNK1 intraventricular injection to investigate its neuroprotective mechanisms.WNK1 expression was significantly reduced in SAH patient blood and SAH model brain tissue,correlating negatively with microglial activation.AAV-WNK1 alleviated brain edema,neuronal necrosis,behavioral deficits,and inflammation by inhibiting NLRP3 inflammasome activation.In hemin-stimulated BV-2 cells,WNK1 overexpression reduced NLRP3 activation and inflammatory cytokines.Chloride counteracted WNK1’s inhibitory effects,and WNK1 suppressed P2X7R-induced NLRP3 activation.Mechanistically,WNK1 functioned via the OXSR1/STK39 pathway.These findings highlight WNK1 as a key regulator of intracellular chloride balance and neuroinflammation,presenting a potential therapeutic target for SAH treatment.
