Neutrophils,macrophages,CD3^(+),CD4^(+),and CD8^(+)T lymphocytes expressμ-,δ-,andκ-opioid receptors(ORs)with varying affinities for opioids.Mast cells express the atypical OR Mas-related G-protein-coupled receptor ...Neutrophils,macrophages,CD3^(+),CD4^(+),and CD8^(+)T lymphocytes expressμ-,δ-,andκ-opioid receptors(ORs)with varying affinities for opioids.Mast cells express the atypical OR Mas-related G-protein-coupled receptor X2(MRGPRX2),which has a low affinity for morphine.Neutrophils and macrophages can synthesize and release endogenous opioid peptides.Activation of ORs enhances the synthesis of proinflammatory cytokines and the production of reactive oxygen species(ROS)in unstimulated leukocytes.Conversely,OR activation reduces proinflammatory cytokine synthesis in stimulated neutrophils and macrophages.Morphine inhibits Toll-like receptor 4(TLR4)expression in macrophages,thereby attenuating inflammation,whereas methadone induces ROS production in mast cells through TLR4 activation.Stimulation of TLR4 triggersβ-endorphin synthesis in macrophages.The production of proinflammatory cytokines and ROS contributes to cardiac reperfusion injury.Importantly,activation ofκ1-andμ-ORs suppresses proinflammatory cytokine production by leukocytes,thereby mitigating inflammatory injury to the heart and other organs.展开更多
目的基于TLR4/NF-κB信号通路探讨枳葛保肝降脂方含药血清对乙醇诱导的人肝LO2细胞炎症损伤的作用及机制。方法将人肝LO2细胞分为正常组(10%正常大鼠血清)、诱导组(10%正常大鼠血清+2.5%无水乙醇)、美他多辛组(10%美他多辛含药血清+2.5...目的基于TLR4/NF-κB信号通路探讨枳葛保肝降脂方含药血清对乙醇诱导的人肝LO2细胞炎症损伤的作用及机制。方法将人肝LO2细胞分为正常组(10%正常大鼠血清)、诱导组(10%正常大鼠血清+2.5%无水乙醇)、美他多辛组(10%美他多辛含药血清+2.5%无水乙醇)和低、中、高浓度枳葛保肝降脂方含药血清组(简称“低、中、高浓度组”)(10.0%、5.0%、2.5%枳葛保肝降脂方含药血清+2.5%无水乙醇)。采用酶联免疫吸附试验法检测各组细胞裂解液中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-18含量;Western blot法检测各组Toll样受体4(TLR4)、髓样分化因子88(MyD88)、NLR家族含pyrin结构域蛋白3(NLRP3)、Caspase-1、磷酸化核因子κB p65亚基/核因子κB p65亚基(p-NF-κBp65/NF-κBp65)蛋白表达水平;RT-qPCR检测各组TLR4、MYD88、NLRP3、Caspase-1 m RNA表达情况。结果与正常组比较,诱导组中TNF-α、IL-1β、IL-18含量,TLR4、My D88、NLRP3、Caspase-1、p-NF-κBp65/NF-κBp65蛋白表达水平,TLR4、MYD88、NLRP3、Caspase-1 m RNA表达量均升高(P<0.01)。与诱导组比较,中、高浓度组TNF-α、IL-1β、IL-18含量,TLR4、My D88、NLRP3、Caspase-1、p-NF-κBp65/NF-κBp65蛋白表达水平,TLR4、MYD88、NLRP3、Caspase-1、p-NF-κBp65/NF-κBp65mRNA表达量均降低(P<0.01)。结论枳葛保肝降脂方含药血清对无水乙醇诱导的人肝LO2细胞炎症损伤具有保护作用,其作用可能通过TLR4/NF-κB信号通路产生。展开更多
基金supported by the Russian Science Foundation(Grant No.23-65-10017 to B.K.K.and M.K.)The Ministry of Science and Higher Education of the Russian Federation(Grant No.122020300042-4 to L.N.M.)supported the preparation of the minichapter titled"Opioids reduce inflammatory injury of the heart".
文摘Neutrophils,macrophages,CD3^(+),CD4^(+),and CD8^(+)T lymphocytes expressμ-,δ-,andκ-opioid receptors(ORs)with varying affinities for opioids.Mast cells express the atypical OR Mas-related G-protein-coupled receptor X2(MRGPRX2),which has a low affinity for morphine.Neutrophils and macrophages can synthesize and release endogenous opioid peptides.Activation of ORs enhances the synthesis of proinflammatory cytokines and the production of reactive oxygen species(ROS)in unstimulated leukocytes.Conversely,OR activation reduces proinflammatory cytokine synthesis in stimulated neutrophils and macrophages.Morphine inhibits Toll-like receptor 4(TLR4)expression in macrophages,thereby attenuating inflammation,whereas methadone induces ROS production in mast cells through TLR4 activation.Stimulation of TLR4 triggersβ-endorphin synthesis in macrophages.The production of proinflammatory cytokines and ROS contributes to cardiac reperfusion injury.Importantly,activation ofκ1-andμ-ORs suppresses proinflammatory cytokine production by leukocytes,thereby mitigating inflammatory injury to the heart and other organs.
基金supported by the grants for Anhui Provincial University Scientific Research Innovation Team(No.2022AH010083)Anhui Provincial Major Science and Technology Project(No.2022AH040215)the Scientific Research and Innovation Team of Bengbu Medical University,China(No.Byycx24018).
文摘本研究旨在探讨脂质运载蛋白2(lipocalin-2,LCN2)基因敲除(LCN2KO)对盲肠结扎与穿孔(cecal ligation and perforation,CLP)诱导的脓毒症小鼠肺损伤和铁死亡的作用,进一步明确Toll样受体4(Toll-like receptor 4,TLR4)/核因子-κB(nuclear factorκB,NF-κB)通路在其中的可能作用。用LCN2KO小鼠建立CLP模型,用ELISA检测小鼠血清中白介素6(interleukin-6,IL-6)和IL-1β含量,用试剂盒检测肺组织丙二醛(malondialdehyde,MDA)和谷胱甘肽(glutathione,GSH)含量,用DHE荧光探针检测肺组织中活性氧(reactive oxygen species,ROS)水平,用Western blot检测肺组织中铁死亡相关蛋白的表达水平。结果显示,相对野生型(wild type,WT)-Sham组,WT-CLP组小鼠肺组织明显受损,血清IL-6和IL-1β含量升高,肺组织MDA含量增加,GSH含量降低,ROS水平显著升高,LCN2和TLR4蛋白表达水平显著上调,p-P65/P65比值显著升高,铁死亡抑制蛋白1(ferroptosis suppressor protein 1,FSP1)和谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)蛋白表达水平显著下调。LCN2KO可以逆转脓毒症小鼠的上述变化。TLR4抑制剂TAK-242和NF-κB抑制剂PDTC均可下调脓毒症小鼠肺组织中LCN2蛋白表达水平;与LCN2KO的作用相似,二者均可逆转脓毒症诱导的铁死亡和肺损伤。以上结果提示,LCN2KO可能通过调控TLR4/NF-κB信号通路减轻铁死亡,从而改善脓毒症小鼠肺损伤。
文摘目的基于TLR4/NF-κB信号通路探讨枳葛保肝降脂方含药血清对乙醇诱导的人肝LO2细胞炎症损伤的作用及机制。方法将人肝LO2细胞分为正常组(10%正常大鼠血清)、诱导组(10%正常大鼠血清+2.5%无水乙醇)、美他多辛组(10%美他多辛含药血清+2.5%无水乙醇)和低、中、高浓度枳葛保肝降脂方含药血清组(简称“低、中、高浓度组”)(10.0%、5.0%、2.5%枳葛保肝降脂方含药血清+2.5%无水乙醇)。采用酶联免疫吸附试验法检测各组细胞裂解液中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-18含量;Western blot法检测各组Toll样受体4(TLR4)、髓样分化因子88(MyD88)、NLR家族含pyrin结构域蛋白3(NLRP3)、Caspase-1、磷酸化核因子κB p65亚基/核因子κB p65亚基(p-NF-κBp65/NF-κBp65)蛋白表达水平;RT-qPCR检测各组TLR4、MYD88、NLRP3、Caspase-1 m RNA表达情况。结果与正常组比较,诱导组中TNF-α、IL-1β、IL-18含量,TLR4、My D88、NLRP3、Caspase-1、p-NF-κBp65/NF-κBp65蛋白表达水平,TLR4、MYD88、NLRP3、Caspase-1 m RNA表达量均升高(P<0.01)。与诱导组比较,中、高浓度组TNF-α、IL-1β、IL-18含量,TLR4、My D88、NLRP3、Caspase-1、p-NF-κBp65/NF-κBp65蛋白表达水平,TLR4、MYD88、NLRP3、Caspase-1、p-NF-κBp65/NF-κBp65mRNA表达量均降低(P<0.01)。结论枳葛保肝降脂方含药血清对无水乙醇诱导的人肝LO2细胞炎症损伤具有保护作用,其作用可能通过TLR4/NF-κB信号通路产生。
