Objective To investigate whether Tuina alleviates fibrotic symptoms in myofascial trigger points(MTrPs)by regulating transforming growth factor(TGF)-β1/Smad3 signaling pathway,thereby deactivating these points.Method...Objective To investigate whether Tuina alleviates fibrotic symptoms in myofascial trigger points(MTrPs)by regulating transforming growth factor(TGF)-β1/Smad3 signaling pathway,thereby deactivating these points.Methods This study comprised two experimental phases.In phase 1,27 specific pathogenfree(SPF)grade female Sprague-Dawley(SD)rats were randomized into three groups:control 1,model 1,and Tuina 1 groups.Model 1 and Tuina 1 groups underwent an 8-week MTrPs modeling protocol involving blunt impact and eccentric exercise.After successful modeling,rats in Tuina 1 group received manual pressing on nodules or cord-like taut bands on the medial aspect of the left hindlimb.Pain sensitivity and tissue stiffness were evaluated via pressure pain threshold(PPT)and soft tissue tension(STT).Muscle histopathology and fibrosis were observed using hematoxylin and eosin(HE)and Masson staining.Inflammatory factors in muscle were measured by enzyme-linked immunosorbent assay(ELISA),while immunofluorescence(IF)and Western blot(WB)were used to detect the expression levels ofα-smooth muscle actin(α-SMA),collagenⅢ,and TGF-β1.In phase 2,45 SPF female SD rats were randomized into five groups:control 2,model 2,Tuina 2,TGF-β1 inhibitor(TI),and Tuina+TGF-β1 agonist(Tuina+TA)groups.All groups except control 2 underwent standardized MTrPs modeling.Rats in Tuina 2 group received consistent pressing manipulation.TI group received intraperitoneal injections of oxymatrine,while Tuina+TA group received intraperitoneal injections of SRI-011381 hydrochloride followed by the same pressing protocol as Tuina 2 group.WB was used to detect the expression of collagen I,collagen III,TGF-β1,and phosphorylated-Smad3(p-Smad3)/Smad3.Results In phase 1,Tuina significantly improved PPT and STT in MTrPs of rats(P<0.01),reversed pathological damages including disorganized muscle fiber arrangement,abnormal myocyte morphology,and exacerbated fibrosis.In addition,in MTrPs of rats in model 1 group,expression levels of nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB),interleukin(IL)-1β,IL-6,tumor necrosis factor(TNF)-α,and fibrosis markers(α-SMA,collagen I,and collagen III)were upregulated,and all exhibited a significant downward trend after Tuina intervention(P<0.05 or P<0.01).This indicates that the therapeutic effects of Tuina are directly associated with reduced local inflammation and fibrosis in MTrPs.In phase 2,compared with model 2 group,rats in TI and Tuina 2 groups had decreased expression levels of TGF-β1 and p-Smad3/Smad3 in MTrPs,alongside reduced levels of inflammatory factors(IL-1β,IL-6,NF-κB,and TNF-α)and fibrosis markers(α-SMA,collagen I,and collagen III)(P<0.05 or P<0.01).When co-administered with TGF-β1 agonist,the therapeutic effects of Tuina were significantly attenuated,with rebounded TGF-β1 expression and p-Smad3/Smad3 in local MTrPs,and fibrosis and inflammatory responses were re-exacerbated(P<0.05 or P<0.01).Conclusion Tuina can effectively reduce inflammatory responses and fibrosis in MTrPs tissue,and its mechanism is closely related to the inhibition of the TGF-β1/Smad3 signaling pathway,which plays a critical role in Tuina-mediated regulation of MTrPs fibrosis.展开更多
Glaucoma is characterized by chronic progressive optic nerve damage and retinal ganglion cell death.Although extensive research has been conducted on neuroprotection for retinal ganglion cells,there is still no treatm...Glaucoma is characterized by chronic progressive optic nerve damage and retinal ganglion cell death.Although extensive research has been conducted on neuroprotection for retinal ganglion cells,there is still no treatment for clinical use.Recent evidence shows that extracellular vesicles isolated from a variety of stem cells are efficacious in retinal ganglion cell neuroprotection.In this study,we tested the novel extracellular vesicle source of the retinal progenitor R-28 cell line in vitro and in vivo.We isolated and characterized extracellular vesicles from R-28 cells and tested their therapeutic efficacy in terms of retinal ganglion cell survival in vitro and in an in vivo glaucoma model,measuring retinal ganglion cell survival and preservation of their axons.Additionally,we tested extracellular vesicles for their neuroprotective capacity in retinal ganglion cells differentiated from human embryonic stem cells.Finally,we investigated miRNA changes in retinal ganglion cells with R-28 extracellular vesicle treatment,and predicted possible pathways that may be modulated.R-28 extracellular vesicles improved retinal ganglion cell survival but failed to preserve axons significantly.Moreover,the results also illustrated the neuroprotection of R-28 extracellular vesicles on human retinal ganglion cells.Finally,we also showed changes in hsa-miRNA-4443,hsa-miRNA-216a-5p,hsa-let-7e-5p,hsa-miRNA-374b-5p,hsa-miRNA-331-3p,and hsa-miRNA-421 expressions,which may have neuroprotective potential on retinal ganglion cell degeneration.This study will pave the way for miRNA and extracellular vesicle-based neuroprotective therapies for glaucoma.展开更多
GABA_(A) receptors containingα5-subunits(GABA_(A)R-α5)cluster at both extrasynaptic and synaptic locations,interacting with the scaffold proteins radixin and gephyrin,respectively,and the re-localization of GABA_(A...GABA_(A) receptors containingα5-subunits(GABA_(A)R-α5)cluster at both extrasynaptic and synaptic locations,interacting with the scaffold proteins radixin and gephyrin,respectively,and the re-localization of GABA_(A)R-α5 influences GABAergic transmission.Here,we found that when early spatial memory deficits occurred in aged mice at 24 h after sevoflurane anesthesia,there was a re-localization of GABA_(A)R-α5 that enhanced tonic inhibition and reduced the decay kinetics of miniature inhibitory postsynaptic currents in the hippocampal CA1 region.Mechanistically,increased phosphorylation of radixin at threonine 564(Thr564)mediates the re-localization of GABA_(A)R-α5.Acute treatment with the selective extrasynaptic GABA_(A)R-α5 antagonist S44819 restored the GABA_(A)R-α5-mediated inhibitory currents by reversing radixin phosphorylation-dependent GABA_(A)R-α5 re-localization,then improved the sevoflurane-induced spatial memory impairment in aged mice.Our results suggest that the localization of GABA_(A)R-α5 altered by sevoflurane is linked to unbalanced GABAergic transmission,which induces early memory impairment in aged mice.Modulating the GABA_(A)R-α5 localization might be a novel strategy to improve memory after sevoflurane exposure.展开更多
基金Natural Science Foundation of China(82274676 and 82374613)Program of Hunan Provincial Natural Science(2023JJ30458).
文摘Objective To investigate whether Tuina alleviates fibrotic symptoms in myofascial trigger points(MTrPs)by regulating transforming growth factor(TGF)-β1/Smad3 signaling pathway,thereby deactivating these points.Methods This study comprised two experimental phases.In phase 1,27 specific pathogenfree(SPF)grade female Sprague-Dawley(SD)rats were randomized into three groups:control 1,model 1,and Tuina 1 groups.Model 1 and Tuina 1 groups underwent an 8-week MTrPs modeling protocol involving blunt impact and eccentric exercise.After successful modeling,rats in Tuina 1 group received manual pressing on nodules or cord-like taut bands on the medial aspect of the left hindlimb.Pain sensitivity and tissue stiffness were evaluated via pressure pain threshold(PPT)and soft tissue tension(STT).Muscle histopathology and fibrosis were observed using hematoxylin and eosin(HE)and Masson staining.Inflammatory factors in muscle were measured by enzyme-linked immunosorbent assay(ELISA),while immunofluorescence(IF)and Western blot(WB)were used to detect the expression levels ofα-smooth muscle actin(α-SMA),collagenⅢ,and TGF-β1.In phase 2,45 SPF female SD rats were randomized into five groups:control 2,model 2,Tuina 2,TGF-β1 inhibitor(TI),and Tuina+TGF-β1 agonist(Tuina+TA)groups.All groups except control 2 underwent standardized MTrPs modeling.Rats in Tuina 2 group received consistent pressing manipulation.TI group received intraperitoneal injections of oxymatrine,while Tuina+TA group received intraperitoneal injections of SRI-011381 hydrochloride followed by the same pressing protocol as Tuina 2 group.WB was used to detect the expression of collagen I,collagen III,TGF-β1,and phosphorylated-Smad3(p-Smad3)/Smad3.Results In phase 1,Tuina significantly improved PPT and STT in MTrPs of rats(P<0.01),reversed pathological damages including disorganized muscle fiber arrangement,abnormal myocyte morphology,and exacerbated fibrosis.In addition,in MTrPs of rats in model 1 group,expression levels of nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB),interleukin(IL)-1β,IL-6,tumor necrosis factor(TNF)-α,and fibrosis markers(α-SMA,collagen I,and collagen III)were upregulated,and all exhibited a significant downward trend after Tuina intervention(P<0.05 or P<0.01).This indicates that the therapeutic effects of Tuina are directly associated with reduced local inflammation and fibrosis in MTrPs.In phase 2,compared with model 2 group,rats in TI and Tuina 2 groups had decreased expression levels of TGF-β1 and p-Smad3/Smad3 in MTrPs,alongside reduced levels of inflammatory factors(IL-1β,IL-6,NF-κB,and TNF-α)and fibrosis markers(α-SMA,collagen I,and collagen III)(P<0.05 or P<0.01).When co-administered with TGF-β1 agonist,the therapeutic effects of Tuina were significantly attenuated,with rebounded TGF-β1 expression and p-Smad3/Smad3 in local MTrPs,and fibrosis and inflammatory responses were re-exacerbated(P<0.05 or P<0.01).Conclusion Tuina can effectively reduce inflammatory responses and fibrosis in MTrPs tissue,and its mechanism is closely related to the inhibition of the TGF-β1/Smad3 signaling pathway,which plays a critical role in Tuina-mediated regulation of MTrPs fibrosis.
基金supported by a Ph.D.scholarship from the YLSY program of the Republic of Turkiye,Ministry of National Educationfunded by Fight for Sight UK,grant reference#5183/5184。
文摘Glaucoma is characterized by chronic progressive optic nerve damage and retinal ganglion cell death.Although extensive research has been conducted on neuroprotection for retinal ganglion cells,there is still no treatment for clinical use.Recent evidence shows that extracellular vesicles isolated from a variety of stem cells are efficacious in retinal ganglion cell neuroprotection.In this study,we tested the novel extracellular vesicle source of the retinal progenitor R-28 cell line in vitro and in vivo.We isolated and characterized extracellular vesicles from R-28 cells and tested their therapeutic efficacy in terms of retinal ganglion cell survival in vitro and in an in vivo glaucoma model,measuring retinal ganglion cell survival and preservation of their axons.Additionally,we tested extracellular vesicles for their neuroprotective capacity in retinal ganglion cells differentiated from human embryonic stem cells.Finally,we investigated miRNA changes in retinal ganglion cells with R-28 extracellular vesicle treatment,and predicted possible pathways that may be modulated.R-28 extracellular vesicles improved retinal ganglion cell survival but failed to preserve axons significantly.Moreover,the results also illustrated the neuroprotection of R-28 extracellular vesicles on human retinal ganglion cells.Finally,we also showed changes in hsa-miRNA-4443,hsa-miRNA-216a-5p,hsa-let-7e-5p,hsa-miRNA-374b-5p,hsa-miRNA-331-3p,and hsa-miRNA-421 expressions,which may have neuroprotective potential on retinal ganglion cell degeneration.This study will pave the way for miRNA and extracellular vesicle-based neuroprotective therapies for glaucoma.
基金supported by the Tianjin Scientific Research Start-up Foundation for Talent Introduction(2021-1-10)the 14th Five-Year Plan Peak Discipline Support Plan of Tianjin Medical University Cancer Institute and Hospital(7-2-13)+3 种基金National Natural Science Foundation of China(82171221),Beijing Bethune Charitable Foundation(YXJL-2024-0778-0030)Beijing Science and Technology Innovation Medical Development Foundation(KC2024-JF-0069)Tianjin Medical University Postgraduate Education Reform Research Program(TMUYY02)Tianjin Key Medical Discipline(Specialty)Construction(TJYXZDXK-009A).
文摘GABA_(A) receptors containingα5-subunits(GABA_(A)R-α5)cluster at both extrasynaptic and synaptic locations,interacting with the scaffold proteins radixin and gephyrin,respectively,and the re-localization of GABA_(A)R-α5 influences GABAergic transmission.Here,we found that when early spatial memory deficits occurred in aged mice at 24 h after sevoflurane anesthesia,there was a re-localization of GABA_(A)R-α5 that enhanced tonic inhibition and reduced the decay kinetics of miniature inhibitory postsynaptic currents in the hippocampal CA1 region.Mechanistically,increased phosphorylation of radixin at threonine 564(Thr564)mediates the re-localization of GABA_(A)R-α5.Acute treatment with the selective extrasynaptic GABA_(A)R-α5 antagonist S44819 restored the GABA_(A)R-α5-mediated inhibitory currents by reversing radixin phosphorylation-dependent GABA_(A)R-α5 re-localization,then improved the sevoflurane-induced spatial memory impairment in aged mice.Our results suggest that the localization of GABA_(A)R-α5 altered by sevoflurane is linked to unbalanced GABAergic transmission,which induces early memory impairment in aged mice.Modulating the GABA_(A)R-α5 localization might be a novel strategy to improve memory after sevoflurane exposure.
