Background:This study investigated the role of polydatin in regulating macrophage-epithelial cell(EC)interactions during asthma.An asthma model was induced in BALB/c mice using ovalbumin(20μg).Methods:The therapeutic...Background:This study investigated the role of polydatin in regulating macrophage-epithelial cell(EC)interactions during asthma.An asthma model was induced in BALB/c mice using ovalbumin(20μg).Methods:The therapeutic effects of polydatin(20 and 40 mg/kg)were evaluated in this asthmatic mouse model.To assess the underlying mechanisms,Bronchial Epithelium Adenovirus 12-SV402B(BEAS-2B)cells were cocultured with Tohoku Hospital for Pediatrics-1(THP-1)macrophages,in which toll-like receptor 4(TLR4)was either overexpressed or knocked down,and subsequently stimulated with lipopoly-saccharide(LPS)and ATP.THP-1 cells underwent a 1-h pretreatment with polydatin(50 and 100μmol/L),Class Lipid Inhibitor-095(CLI-095,TLR4 inhibitor,1μg/mL),or A438079(P2X7R antagonist,10μmol/L)prior to LPS/ATP challenge.Results:Findings from Western blotting,enzyme-linked immunosorbent assay,flow cytometry,real-time polymerase chain reaction,and immunofluorescence assays demonstrated that modulating TLR4 expression significantly altered interleukin-1β(IL-1β)secretion from THP-1 macrophages and mitochondrial reactive oxygen species(mtROS)production in BEAS-2B ECs.In the mouse asthma model,polydatin significantly alleviated airway inflammation,oxidative stress,and apoptosis,likely by interfering with TLR4/P2X7R-mediated signaling and suppressing the activation of the NOD-like receptor protein inflammasome.Additionally,polydatin significantly reduced IL-1βand IL-18 levels and inhibited the infiltration of macrophages and eosinophils.Correspondingly,polydatin significantly attenuated TLR4/P2X7R signaling in THP-1 cells stimulated with ATP and LPS,thereby reducing IL-1βand IL-18 secretion,calcium influx,mtROS production,and apoptosis in BEAS-2B ECs.Conclusions:Polydatin is a promising therapeutic candidate for asthma,possibly by targeting macrophage-epithelium cross-talk via the TLR4/P2X7R axis.Future formulations as capsules or sprays may effectively alleviate airway inflammation and remodeling.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
Dysfunctional autophagy often occurs during the development of neurodegenerative diseases,such as Parkinson’s disease,Huntington’s disease,and Alzheimer’s disease.The purinergic P2X4 receptor is an ATP-gated ion ch...Dysfunctional autophagy often occurs during the development of neurodegenerative diseases,such as Parkinson’s disease,Huntington’s disease,and Alzheimer’s disease.The purinergic P2X4 receptor is an ATP-gated ion channel that is widely expressed in the microglia,astrocytes,and neurons of the central and peripheral nervous systems.P2X4R is involved in the regulation of cellular excitability,synaptic transmission,and neuroinflammation.However,the role played by P2X4R in Parkinson’s disease remains poorly understood.Rat models of Parkinson’s disease were established by injecting 6-hydroxydopamine into the substantia nigra pars compacta.P2X4R-targeted small interfering RNA(siRNA)was injected into the same area 1 week before injury induction to inhibit the expression of the P2X4 receptor.The results showed that the inhibition of P2X4 receptor expression in Parkinson’s disease model rats reduced the rotation behavior induced by apomorphine treatment,increased the latency on the rotarod test,and upregulated the expression of tyrosine hydroxylase,brain-derived neurotrophic factor,LC3-II/LC3-I,Beclin-1,and phosphorylated tropomyosin receptor kinase B(TrkB)in brain tissue,while simultaneously reducing p62 levels.These findings suggest that P2X4 receptor activation might inhibit neuronal autophagy through the regulation of the brain-derived neurotrophic factor/TrkB signaling pathway,leading to dopaminergic neuron damage in the substantia nigra and the further inhibition of P2X4 receptor-mediated autophagy.These results indicate that P2X4 receptor might serve as a potential novel target for the treatment of Parkinson’s disease.This study was approved by the Animal Ethics Committee of Affiliated Hospital of Qingdao University(approval No.QYFYWZLL26119)on April 12,2016.展开更多
基金National Natural Science Foundation of China,Grant/Award Number:82260007Jilin Province Health Commission,Grant/Award Number:2024A062+1 种基金Jilin Provincial Department of Education,Grant/Award Number:JJKH20240698KJJilin Province Science and Technology Department,Grant/Award Number:20240404025ZP and 20240602100RC。
文摘Background:This study investigated the role of polydatin in regulating macrophage-epithelial cell(EC)interactions during asthma.An asthma model was induced in BALB/c mice using ovalbumin(20μg).Methods:The therapeutic effects of polydatin(20 and 40 mg/kg)were evaluated in this asthmatic mouse model.To assess the underlying mechanisms,Bronchial Epithelium Adenovirus 12-SV402B(BEAS-2B)cells were cocultured with Tohoku Hospital for Pediatrics-1(THP-1)macrophages,in which toll-like receptor 4(TLR4)was either overexpressed or knocked down,and subsequently stimulated with lipopoly-saccharide(LPS)and ATP.THP-1 cells underwent a 1-h pretreatment with polydatin(50 and 100μmol/L),Class Lipid Inhibitor-095(CLI-095,TLR4 inhibitor,1μg/mL),or A438079(P2X7R antagonist,10μmol/L)prior to LPS/ATP challenge.Results:Findings from Western blotting,enzyme-linked immunosorbent assay,flow cytometry,real-time polymerase chain reaction,and immunofluorescence assays demonstrated that modulating TLR4 expression significantly altered interleukin-1β(IL-1β)secretion from THP-1 macrophages and mitochondrial reactive oxygen species(mtROS)production in BEAS-2B ECs.In the mouse asthma model,polydatin significantly alleviated airway inflammation,oxidative stress,and apoptosis,likely by interfering with TLR4/P2X7R-mediated signaling and suppressing the activation of the NOD-like receptor protein inflammasome.Additionally,polydatin significantly reduced IL-1βand IL-18 levels and inhibited the infiltration of macrophages and eosinophils.Correspondingly,polydatin significantly attenuated TLR4/P2X7R signaling in THP-1 cells stimulated with ATP and LPS,thereby reducing IL-1βand IL-18 secretion,calcium influx,mtROS production,and apoptosis in BEAS-2B ECs.Conclusions:Polydatin is a promising therapeutic candidate for asthma,possibly by targeting macrophage-epithelium cross-talk via the TLR4/P2X7R axis.Future formulations as capsules or sprays may effectively alleviate airway inflammation and remodeling.
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.
基金This work was supported by the National Natural Science Foundation of China,Nos.81571225 and 81971192(both to AMX).
文摘Dysfunctional autophagy often occurs during the development of neurodegenerative diseases,such as Parkinson’s disease,Huntington’s disease,and Alzheimer’s disease.The purinergic P2X4 receptor is an ATP-gated ion channel that is widely expressed in the microglia,astrocytes,and neurons of the central and peripheral nervous systems.P2X4R is involved in the regulation of cellular excitability,synaptic transmission,and neuroinflammation.However,the role played by P2X4R in Parkinson’s disease remains poorly understood.Rat models of Parkinson’s disease were established by injecting 6-hydroxydopamine into the substantia nigra pars compacta.P2X4R-targeted small interfering RNA(siRNA)was injected into the same area 1 week before injury induction to inhibit the expression of the P2X4 receptor.The results showed that the inhibition of P2X4 receptor expression in Parkinson’s disease model rats reduced the rotation behavior induced by apomorphine treatment,increased the latency on the rotarod test,and upregulated the expression of tyrosine hydroxylase,brain-derived neurotrophic factor,LC3-II/LC3-I,Beclin-1,and phosphorylated tropomyosin receptor kinase B(TrkB)in brain tissue,while simultaneously reducing p62 levels.These findings suggest that P2X4 receptor activation might inhibit neuronal autophagy through the regulation of the brain-derived neurotrophic factor/TrkB signaling pathway,leading to dopaminergic neuron damage in the substantia nigra and the further inhibition of P2X4 receptor-mediated autophagy.These results indicate that P2X4 receptor might serve as a potential novel target for the treatment of Parkinson’s disease.This study was approved by the Animal Ethics Committee of Affiliated Hospital of Qingdao University(approval No.QYFYWZLL26119)on April 12,2016.
