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
Recent studies have shown that fibrotic scar formation following cerebral ischemic injury has varying effects depending on the microenvironment.However,little is known about how fibrosis is induced and regulated after...Recent studies have shown that fibrotic scar formation following cerebral ischemic injury has varying effects depending on the microenvironment.However,little is known about how fibrosis is induced and regulated after cerebral ischemic injury.Sonic hedgehog signaling participates in fibrosis in the heart,liver,lung,and kidney.Whether Shh signaling modulates fibrotic scar formation after cerebral ischemic stroke and the underlying mechanisms are unclear.In this study,we found that Sonic Hedgehog expression was upregulated in patients with acute ischemic stroke and in a middle cerebral artery occlusion/reperfusion injury rat model.Both Sonic hedgehog and Mitofusin 2 showed increased expression in the middle cerebral artery occlusion rat model and in vitro fibrosis cell model induced by transforming growth factor-beta 1.Activation of the Sonic hedgehog signaling pathway enhanced the expression of phosphorylated Smad 3 and Mitofusin 2 proteins,promoted the formation of fibrotic scars,protected synapses or promoted synaptogenesis,alleviated neurological deficits following middle cerebral artery occlusion/reperfusion injury,reduced cell apoptosis,facilitated the transformation of meninges fibroblasts into myofibroblasts,and enhanced the proliferation and migration of meninges fibroblasts.The Smad3 phosphorylation inhibitor SIS3 reversed the effects induced by Sonic hedgehog signaling pathway activation.Bioinformatics analysis revealed significant correlations between Sonic hedgehog and Smad3,between Sonic hedgehog and Mitofusin 2,and between Smad3 and Mitofusin 2.These findings suggest that Sonic hedgehog signaling may influence Mitofusin 2 expression by regulating Smad3 phosphorylation,thereby modulating the formation of early fibrotic scars following cerebral ischemic stroke and affecting prognosis.The Sonic Hedgehog signaling pathway may serve as a new therapeutic target for stroke treatment.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 the National Natural Science Foundation of China,Nos.82171456(to QY)and 81971229(to QY)the Natural Science Foundation of Chongqing,Nos.CSTC2021JCYJ-MSXMX0263(to QY)and CSTB2023NSCQ-MSX1015(to XL)Doctoral Innovation Project of The First Affiliated Hospital of Chongqing Medical University,Nos.CYYY-BSYJSCXXM-202318(to JW)and CYYY-BSYJSCXXM-202327(to HT).
文摘Recent studies have shown that fibrotic scar formation following cerebral ischemic injury has varying effects depending on the microenvironment.However,little is known about how fibrosis is induced and regulated after cerebral ischemic injury.Sonic hedgehog signaling participates in fibrosis in the heart,liver,lung,and kidney.Whether Shh signaling modulates fibrotic scar formation after cerebral ischemic stroke and the underlying mechanisms are unclear.In this study,we found that Sonic Hedgehog expression was upregulated in patients with acute ischemic stroke and in a middle cerebral artery occlusion/reperfusion injury rat model.Both Sonic hedgehog and Mitofusin 2 showed increased expression in the middle cerebral artery occlusion rat model and in vitro fibrosis cell model induced by transforming growth factor-beta 1.Activation of the Sonic hedgehog signaling pathway enhanced the expression of phosphorylated Smad 3 and Mitofusin 2 proteins,promoted the formation of fibrotic scars,protected synapses or promoted synaptogenesis,alleviated neurological deficits following middle cerebral artery occlusion/reperfusion injury,reduced cell apoptosis,facilitated the transformation of meninges fibroblasts into myofibroblasts,and enhanced the proliferation and migration of meninges fibroblasts.The Smad3 phosphorylation inhibitor SIS3 reversed the effects induced by Sonic hedgehog signaling pathway activation.Bioinformatics analysis revealed significant correlations between Sonic hedgehog and Smad3,between Sonic hedgehog and Mitofusin 2,and between Smad3 and Mitofusin 2.These findings suggest that Sonic hedgehog signaling may influence Mitofusin 2 expression by regulating Smad3 phosphorylation,thereby modulating the formation of early fibrotic scars following cerebral ischemic stroke and affecting prognosis.The Sonic Hedgehog signaling pathway may serve as a new therapeutic target for stroke treatment.
