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Spleen tyrosine kinase (Syk) is an intracellular tyrosine kinase involved in the signal transduction in immune cells mainly. Its aberrant regulation is associated with diversified allergic disorders, autoimmune diseases and B cell malignancies. Therefore, inhibition of Syk is considered a reasonable approach to treat autoimmune/inflammatory diseases and B cell malignancies. Here we described the preclinical characterization of sovleplenib, a novel, highly potent and selective, oral Syk inhibitor, in several rodent autoimmune disease models. Sovleplenib potently inhibited Syk activity in a recombinant enzymatic assay and Syk-dependent cellular functions in various immune cell lines and human whole blood in vitro. Furthermore, sovleplenib, by oral administration, demonstrated strong in vivo efficacies in murine models of immune thrombocytopenia (ITP), autoimmune hemolytic anemia (AIHA), and chronic graft-versus-host disease (cGVHD), and a rat model of collagen induced arthritis (CIA) respectively, in a dose-dependent manner. Collectively, these results clearly supported sovleplenib as a therapeutic agent in the treatment of autoimmune diseases. Sovleplenib is being globally developed for ITP (Phase III, NCT05029635, Phase Ib/II, NCT03951623), wAIHA (Phase II/III, NCT05535933) and B-cell lymphoma (Phase I, NCT02857998, NCT03779113). SIGNIFICANCE STATEMENT: Syk is a key mediator of signaling pathways downstream of a wide array of receptors important for immune functions, including the B cell receptor, immunoglobulin receptors bearing Fc receptors. Inhibition of Syk could provide a novel therapeutic approach for autoimmune diseases and hematologic malignancies. The manuscript describes the preclinical pharmacology characterization of sovleplenib, a novel Syk inhibitor, in enzymatic and cellular assays in vitro and several murine autoimmune disease models in vivo.
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Doenças Autoimunes , Neoplasias , Ratos , Camundongos , Humanos , Animais , Proteínas Tirosina Quinases , Quinase Syk , Transdução de Sinais , Inibidores de Proteínas Quinases/farmacologia , Doenças Autoimunes/tratamento farmacológico , Neoplasias/tratamento farmacológicoRESUMO
In regions reliant on fisheries for livelihoods, a significant number of fish by-products are generated annually due to processing. These discarded parts contain valuable biological resources, such as proteins, fish oils, and trace elements, thus holding enormous potential for reutilization. In recent years, fish by-product proteins have been widely utilized in skincare products due to their rich collagen content, biosafety, and biocompatibility. This review summarizes the research into and applications of fish by-product proteins in skin health, including alleviating oxidative stress and skin inflammation, reducing DNA damage, mitigating melanin production, improving skin hydration, slowing skin matrix degradation, and promoting synthesis. Additionally, the possibility of improving skin health by improving the abundance of gut microbiota is also discussed. This review underscores the importance of fish by-product proteins in the fisheries, food processing, cosmetics, and biomedical industries.
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Proteínas de Peixes , Pele , Animais , Humanos , Pele/metabolismo , Pele/efeitos dos fármacos , Proteínas de Peixes/metabolismo , Peixes/metabolismo , Cosméticos , Estresse Oxidativo/efeitos dos fármacosRESUMO
BACKGROUND: Savolitinib has shown good tolerability and preliminary efficacy, but efficacy biomarkers require investigation. The main purpose of this study was to confirm in Chinese patients the recommended phase II dose (RP2D) of savolitinib and to explore overall benefit in tumors bearing c-Met aberration. METHODS: This was an open-label, multi-center, 2-part phase I study. A starting dose of 600 mg QD was initiated in the escalation phase, utilizing a 3+3 design with repeated QD and BID dosing. In the dose expansion phase, we enrolled patients with gastric cancer and non-small cell lung cancer (NSCLC) with documented c-met aberration into 5 cohorts to further explore biomarkers. c-Met overexpression and amplification were assessed by immunohistochemistry and FISH, respectively. RESULTS: The safety analysis set included 85 patients. Only one dose-limiting toxicity (grade 3 fatigue) was reported in the 600 mg BID dosing group. The most frequent treatment-related adverse events were nausea (29.4%), vomiting (27.1%), and peripheral edema (21.2%). Notably, in gastric cancer, response was only observed in patients with MET amplification (copy number 9.7-18.4), with an objective response rate of 35.7% and a disease control rate of 64.3%. For patients with NSCLC bearing a MET exon 14 skipping mutation, obvious target lesion shrinkage was observed in 2 of 4 patients, although PR was not achieved. CONCLUSION: The RP2D of savolitinib was established as 600 mg QD or 500 mg BID in Chinese patients. The promising response observed in patients with gastric cancer with c-met amplification and NSCLC with MET exon 14 skipping mutation warrants further investigation. CLINICALTRIALS.GOV IDENTIFIER: NCT0198555.
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The transcriptional repressor Bmi-1 is involved in cell-cycle regulation and cell senescence, the deficiency of which has been shown to cause oxidative stress. This study investigated whether Bmi-1 deficiency plays a role in promoting disc degeneration and the effect of treatment with antioxidant N-acetylcysteine (NAC) on intervertebral disc degeneration. Bmi-1-/- mice were treated with the antioxidant NAC, supplied in drinking water (Bmi-1-/- +NAC). For in vitro experiments, mouse intervertebral discs were cultured under low oxygen tension and serum-limiting conditions in the presence of tumour necrosis factor α and interleukin 1ß in order to mimic degenerative insult. Disc metabolism parameters in these in vitro and in vivo studies were evaluated by histopathological, immunohistochemical and molecular methods. Bmi-1-/- mice showed lower collagen â ¡ and aggrecan levels and higher collagen â © levels than wild-type and Bmi-1-/- +NAC mice. Bmi-1-/- mice showed significantly lower superoxide dismutase (SOD)-1, SOD-2, glutathione peroxidase (GPX)-1 and GPX-3 levels than their wild-type littermates and Bmi-1-/- + NAC mice. Relative to Bmi-1-/- mice, the control and Bmi-1-/- +NAC mice showed significantly lower p16, p21, and p53 levels. These results demonstrate that Bmi-1 plays an important role in attenuating intervertebral disc degeneration in mice by inhibiting oxidative stress and cell apoptosis.
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Antioxidantes/fisiologia , Degeneração do Disco Intervertebral/tratamento farmacológico , Degeneração do Disco Intervertebral/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Complexo Repressor Polycomb 1/deficiência , Proteínas Proto-Oncogênicas/deficiência , Acetilcisteína/farmacologia , Agrecanas/metabolismo , Animais , Apoptose/efeitos dos fármacos , Colágeno/metabolismo , Interleucina-1beta/metabolismo , Disco Intervertebral/efeitos dos fármacos , Disco Intervertebral/metabolismo , Camundongos , Técnicas de Cultura de Órgãos/métodos , Superóxido Dismutase/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
In the present study, we investigated whether high dietary Ca and exogenous parathyroid hormone 1-34 fragments (PTH 1-34) have synergistic effects on bone formation in adult mice, and explored the related mechanisms. Adult male mice were fed a normal diet, a high-Ca diet, a PTH-treated diet, or a high-Ca diet combined with subcutaneously injected PTH 1-34 (80 µg/kg per d) for 4 weeks. Bone mineral density, trabecular bone volume, osteoblast number, alkaline phosphatase (ALP)- and type I collagen-positive areas, and the expression levels of osteoblastic bone formation-related genes and proteins were increased significantly in mice fed the high-Ca diet, the PTH-treated diet, and, even more dramatically, the high-Ca diet combined with PTH. Osteoclast number and surface and the ratio of receptor activator for nuclear factor-κB ligand (RANKL):osteoprotegerin (OPG) were decreased in the high-Ca diet treatment group, increased in the PTH treatment group, but not in the combined treatment group. Furthermore, third-passage osteoblasts were treated with high Ca (5 mM), PTH 1-34 (10â»8 M) or high Ca combined with PTH 1-34. Osteoblast viability and ALP activity were increased in either the high Ca-treated or PTH-treated cultures and, even more dramatically, in the cultures treated with high Ca plus PTH, with consistent up-regulation of the expression levels of osteoblast proliferation and differentiation-related genes and proteins. These results indicate that dietary Ca and PTH play synergistic roles in promoting osteoblastic bone formation by stimulating osteoblast proliferation and differentiation.
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Conservadores da Densidade Óssea/uso terapêutico , Reabsorção Óssea/prevenção & controle , Cálcio da Dieta/uso terapêutico , Interações Alimento-Droga , Osteoblastos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Hormônio Paratireóideo/análogos & derivados , Hormônio Paratireóideo/uso terapêutico , Animais , Densidade Óssea/efeitos dos fármacos , Conservadores da Densidade Óssea/farmacologia , Reabsorção Óssea/metabolismo , Reabsorção Óssea/patologia , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/metabolismo , Osso e Ossos/patologia , Canais de Cálcio/genética , Canais de Cálcio/metabolismo , Cálcio da Dieta/metabolismo , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Terapia Combinada , Masculino , Camundongos Endogâmicos C57BL , Osteoblastos/metabolismo , Osteoblastos/patologia , Hormônio Paratireóideo/farmacologia , Fragmentos de Peptídeos/farmacologia , Fragmentos de Peptídeos/uso terapêutico , Receptores de Detecção de Cálcio/agonistas , Receptores de Detecção de Cálcio/genética , Receptores de Detecção de Cálcio/metabolismo , Receptores de Hormônios Paratireóideos/agonistas , Receptores de Hormônios Paratireóideos/genética , Receptores de Hormônios Paratireóideos/metabolismo , Canais de Cátion TRPV/agonistas , Canais de Cátion TRPV/genética , Canais de Cátion TRPV/metabolismo , Regulação para Cima/efeitos dos fármacosRESUMO
BACKGROUND: We compared the perioperative results and complications associated with PLIF and TLIF, and collected evidence for choosing the better fusion method. METHODS: A literature survey of the MEDLINE and EMBASE databases identified 7 comparative observational studies that met our inclusion criteria. Checklists by Cowley were used to evaluate the risk of bias of the included studies. A database including patient demographic information, perioperative results, and complications was established. The summary odds ratio and weighed mean difference with 95% confidence interval were calculated with a random-effects model. RESULTS: We found that PLIF had a higher complication rate (P <0.00001), and TLIF reduced the rate of durotomy (P = 0.01). No statistical difference was found between the two groups with regard to clinical satisfaction (P = 0.54), blood loss (P = 0.14), vertebral root injury (P = 0.08), graft malposition (P = 0.06), infection (P = 0.36), or rate of radiographic fusion (P = 0.27). The evidence indicated that PLIF required longer operative time (P = 0.03). CONCLUSIONS: The evidence indicated that TLIF could reduce the complication rate and durotomy. Neither TLIP nor PLIF was found superior in terms of clinical satisfaction or radiographic fusion rate. PLIF might result in longer time in surgery.
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Vértebras Lombares/cirurgia , Doenças da Coluna Vertebral/cirurgia , Fusão Vertebral/métodos , Ensaios Clínicos como Assunto/métodos , Humanos , Vértebras Lombares/diagnóstico por imagem , Complicações Pós-Operatórias/diagnóstico por imagem , Complicações Pós-Operatórias/etiologia , Radiografia , Doenças da Coluna Vertebral/diagnóstico por imagem , Fusão Vertebral/efeitos adversosRESUMO
OBJECTIVE: To investigate the effects of astaxanthin (AST) on mouse osteoarthritis (OA) and lipopolysaccharide (LPS)-induced ATDC5 cell damage and to explore whether SIRT1 protein plays a role. METHODS: In this study, some mouse OA models were constructed by anterior cruciate ligament transection (ACLT). Imaging, molecular biology and histopathology methods were used to study the effect of AST administration on traumatic OA in mice. In addition, LPS was used to stimulate ATDC5 cells to mimic the inflammatory response of OA. The effects of AST on the cell activity, inflammatory cytokines, matrix metalloproteinases and collagen type II levels were studied by CCK8 activity assay, reverse transcription polymerase chain reaction and protein imprinting. The role of SIRT1 protein was also detected. RESULTS: In the mouse OA model, the articular surface collapsed, the articular cartilage thickness and cartilage matrix protein abundance were significantly decreased, while the expression of inflammatory cytokines and matrix metalloproteinases was increased; but AST treatment reversed these effects. Meanwhile, AST pretreatment could partially reverse LPS-induced ATDC5 cell damage and upregulate SIRT1 expression, but this protective effect of AST was attenuated by concurrent administration of the SIRT1 inhibitor Ex527. CONCLUSION: AST can protect against the early stages of OA by affecting SIRT1 signalling, suggesting that AST might be a potential therapeutic agent for OA treatment.
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Sirtuína 1 , Regulação para Cima , Xantofilas , Xantofilas/farmacologia , Xantofilas/uso terapêutico , Sirtuína 1/metabolismo , Animais , Camundongos , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Modelos Animais de Doenças , Osteoartrite/tratamento farmacológico , Osteoartrite/metabolismo , Osteoartrite/patologia , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/metabolismo , Cartilagem Articular/patologia , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Condrócitos/patologia , Masculino , FenótipoRESUMO
The brain-derived neurotrophic factor (BDNF) participates in the regulation of cortical neurons by influencing the release of glutamate. However, the specific mechanisms are unclear. Hence, we isolated and cultured the cortical neurons of Sprague Dawley rats. Specific inhibitors of TrkB, Src, PLC-γ1, Akt, and MEK1/2 (i.e., K252a, PP2, U73122, LY294002, and PD98059, respectively) were used to treat cortical neurons and to detect the glutamate release from cortical neurons stimulated with BDNF. BDNF significantly increased glutamate release, and simultaneously enhanced phosphorylation levels of TrkB, Src, PLC-γ, Akt, and Erk1/2. For BDNF-stimulated cortical neurons, K252a inhibited glutamate release and inhibited the phosphorylation levels of TrkB, Src, PLC-γ, Erk1/2, and Akt (P < 0.05). PP2 reduced the glutamate release from BDNF-stimulated cortical neurons (P < 0.05) and inhibited the phosphorylation levels of TrkB and PLC-γ1 (P < 0.05). However, PP2 had no effect on the phosphorylation levels of Erk1/2 or Akt (P > 0.05). U73122 inhibited the glutamate release from BDNF-stimulated cortical neurons, but had no influence on the phosphorylation levels of TrkB, Src, Erk1/2, or Akt (P > 0.05). LY294002 and PD98059 did not affect the BDNF-stimulated glutamate release and did not inhibit the phosphorylation levels of TrkB, Src, or PLC-γ1. In summary, BDNF stimulated the glutamate release from cortical neurons via the TrkB/Src/PLC-γ1 signaling pathway.
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Fator Neurotrófico Derivado do Encéfalo/metabolismo , Córtex Cerebral/metabolismo , Ácido Glutâmico/metabolismo , Neurônios/metabolismo , Animais , Fator Neurotrófico Derivado do Encéfalo/antagonistas & inibidores , Fator Neurotrófico Derivado do Encéfalo/genética , Carbazóis/farmacologia , Córtex Cerebral/citologia , Córtex Cerebral/efeitos dos fármacos , Cromonas/farmacologia , Embrião de Mamíferos , Inibidores Enzimáticos/farmacologia , Estrenos/farmacologia , Flavonoides/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Ácido Glutâmico/biossíntese , Alcaloides Indólicos/farmacologia , Morfolinas/farmacologia , Neurônios/citologia , Neurônios/efeitos dos fármacos , Fosfolipase C gama/antagonistas & inibidores , Fosfolipase C gama/genética , Fosfolipase C gama/metabolismo , Fosforilação/efeitos dos fármacos , Cultura Primária de Células , Pirimidinas/farmacologia , Pirrolidinonas/farmacologia , Ratos , Ratos Sprague-Dawley , Receptor trkB/antagonistas & inibidores , Receptor trkB/genética , Receptor trkB/metabolismo , Transdução de Sinais/efeitos dos fármacos , Quinases da Família src/antagonistas & inibidores , Quinases da Família src/genética , Quinases da Família src/metabolismoRESUMO
Osteosarcoma (OS) is one of the most common malignant bone tumors in early adolescence. Multi-drug chemotherapy has greatly increased the five year survival rate from 20% to 70%. However, the rate has been staggering for 30 years and the prognosis is particularly poor for patients with recurrence and metastasis. Our study aimed to investigate the role of Wnt-ß-catenin, Notch and Hedgehog pathway in OS development because all these pathways are involved in skeletal development, tumorigenesis and chemoresistance. Our results showed that the major components in Wnt-ß-catenin pathway, e.g. Wnt3a, ß-catenin and Lef1, were consistently upregulated in human osteosarcoma cell line Saos2 cells compared to human fetal osteoblasts (hFOB), whereas the changes in the expression levels of Notch and Hh signaling molecules were not consistent. Knocking down ß-catenin increased the Saos2 sensitivity to methotrexate (MTX) induced cell death. Consistently, the expression level of ß-catenin protein correlated with the invasiveness of OS, as evidenced by more intensive ß-catenin immunoreactivity in higher grade OS samples. Chemical inhibition of the Wnt-ß-catenin signaling enhanced MTX mediated death of Saos2 cells. A synergistic effect with MTX was observed when both inhibitors for Wnt-ß-catenin and Notch pathways were simultaneously used, while the addition of the Hh inhibitor did not further improve the efficacy. Our findings provide some novel insight to OS pathogenesis and lay a foundation for future application of Wnt-ß-catenin and Notch inhibitors together with the currently used chemotherapeutic drugs to improve the outcome of OS treatment.
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Neoplasias Ósseas/tratamento farmacológico , Resistencia a Medicamentos Antineoplásicos , Osteossarcoma/tratamento farmacológico , Receptores Notch/antagonistas & inibidores , Proteínas Wnt/antagonistas & inibidores , beta Catenina/antagonistas & inibidores , Neoplasias Ósseas/patologia , Linhagem Celular Tumoral , Proteínas Hedgehog/metabolismo , Humanos , Osteossarcoma/patologia , Receptores Notch/metabolismo , Transdução de Sinais , Proteínas Wnt/genética , Proteínas Wnt/metabolismo , beta Catenina/genéticaRESUMO
Objective: To review the mechanism of extracellular vesicles (EVs) in treating intervertebral disc degeneration (IVDD). Methods: The literature about EVs was reviewed and the biological characteristics and mechanism of EVs in the treatment of IVDD were summarized. Results: EVs are a kind of nano-sized vesicles with a double-layered lipid membrane structure secreted by many types of cells. EVs contain many bioactive molecules and participate in the exchange of information between cells, thus they play important roles in inflammation, oxidative stress, senescence, apoptosis, and autophagy. Moreover, EVs are found to slow down the process of IVDD by delaying the pathological progression of the nucleus pulposus, cartilage endplates, and annulus fibrosus. Conclusion: EVs is expected to become a new strategy for the treatment of IVDD, but the specific mechanism remains to be further studied.
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Anel Fibroso , Vesículas Extracelulares , Degeneração do Disco Intervertebral , Disco Intervertebral , Núcleo Pulposo , Humanos , Degeneração do Disco Intervertebral/terapia , Anel Fibroso/patologia , Núcleo Pulposo/patologia , Apoptose , Vesículas Extracelulares/patologiaRESUMO
Background: It has been demonstrated that vitamin D deficiency is associated with an increased risk of patients developing lumbar disc herniation. However, intervertebral disc degeneration caused by active vitamin D deficiency has not been reported. Thus, the purpose of this study was to e investigate the role and mechanism of 1,25-dihydroxyvitamin D (1,25(OH)2D) insufficiency in promoting intervertebral disc degeneration. Methods: The phenotypes of intervertebral discs were compared in wild-type mice and mice with heterozygous deletion of 1α-hydroxylase [1α(OH)ase+/-] at 8 mouths of age using iconography, histology and molecular biology. A mouse model that overexpressed Sirt1 in mesenchymal stem cells on a 1α(OH)ase+/- background (Sirt1Tg/1α(OH)ase+/-) was generated by crossing Prx1-Sirt1 transgenic mice with 1α(OH)ase+/- mice and comparing their intervertebral disc phenotypes with those of Sirt1Tg, 1α(OH)ase+/- and wild-type littermates at 8 months of age. A vitamin D receptor (VDR)-deficient cellular model was generated by knock-down of endogenous VDR using Ad-siVDR transfection into nucleus pulposus cells; VDR-deficient nucleus pulposus cells were then treated with or without resveratrol. The interactions between Sirt1 and acetylated p65, and p65 nuclear localization, were examined using co-immunoprecipitation, Western blots and immunofluorescence staining. VDR-deficient nucleus pulposus cells were also treated with 1,25(OH)2D3, or resveratrol or 1,25(OH)2D3 plus Ex527 (an inhibitor of Sirt1). Effects on Sirt1 expression, cell proliferation, cell senescence, extracellular matrix protein synthesis and degradation, nuclear factor-κB (NF-κB), and expression of inflammatory molecules, were examined, using immunofluorescence staining, Western blots and real-time RT-PCR. Results: 1,25(OH)2D insufficiency accelerated intervertebral disc degeneration by reducing extracellular matrix protein synthesis and enhancing extracellular matrix protein degradation with reduced Sirt1 expression in nucleus pulposus tissues. Overexpression of Sirt1 in MSCs protected against 1,25(OH)2D deficiency-induced intervertebral disc degeneration by decreasing acetylation and phosphorylation of p65 and inhibiting the NF-κB inflammatory pathway. VDR or resveratrol activated Sirt1 to deacetylate p65 and inhibit its nuclear translocation into nucleus pulposus cells. Knockdown of VDR decreased VDR expression and significantly reduced the proliferation and extracellular matrix protein synthesis of nucleus pulposus cells, significantly increased the senescence of nucleus pulposus cells and significantly downregulated Sirt1 expression, and upregulated matrix metallopeptidase 13 (MMP13), tumor necrosis factor-α (TNF-α) and interleukin 1ß (IL-1ß) expression; the ratios of acetylated and phosphorylated p65/p65 in nucleus pulposus cells were also increased. Treatment of nucleus pulposus cells with VDR reduction using 1,25(OH)2D3 or resveratrol partially rescued the degeneration phenotypes, by up-regulating Sirt1 expression and inhibiting NF-κB inflammatory pathway; these effects in nucleus pulposus cells were blocked by inhibition of Sirt1. Conclusion: Results from this study indicate that the 1,25(OH)2D/VDR pathway can prevent the degeneration of nucleus pulposus cells by inhibiting the NF-κB inflammatory pathway mediated by Sirt1.The Translational Potential of This Article: This study provides new insights into the use of 1,25(OH)2D3 to prevent and treat intervertebral disc degeneration caused by vitamin D deficiency.
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Disruption of the blood-spinal cord barrier (BSCB) leads to inflammatory cell infiltration and neural cell death, thus, contributing to poor functional recovery after spinal cord injury (SCI). Previous studies have suggested that Sirtuin 1 (SIRT1), an NAD+-dependent class III histone deacetylase, is abundantly expressed in endothelial cells and promotes endothelial homeostasis. However, the role of SIRT1 in BSCB function after SCI remains poorly defined. Here, we report that SIRT1 is highly expressed in spinal cord endothelial cells, and its expression significantly decreases after SCI. Using endothelial cell-specific SIRT1 knockout mice, we observed that endothelial cell-specific knockout of SIRT1 aggravated BSCB disruption, thus, resulting in widespread inflammation, neural cell death and poor functional recovery after SCI. In contrast, activation of SIRT1 by the agonist SRT1720 had beneficial effects. In vitro, knockdown of SIRT1 exacerbated IL-1ß-induced endothelial barrier disruption in bEnd.3 cells, whereas overexpression of SIRT1 was protective. Using RNA-seq and IP/MS analysis, we identified p66Shc, a redox protein, as the potential target of SIRT1. Further studies demonstrated that SIRT1 interacts with and deacetylates p66Shc, thereby attenuating oxidative stress and protecting endothelial barrier function. Overall, our results indicate that SIRT1 decreases endothelial ROS production and attenuates BSCB disruption by deacetylating p66Shc after SCI, and suggest that SIRT1 activation has potential as a therapeutic approach to promote functional recovery against BSCB disruption following SCI.
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Sirtuína 1 , Traumatismos da Medula Espinal , Animais , Camundongos , Barreira Hematoencefálica , Células Endoteliais/metabolismo , Camundongos Knockout , Sirtuína 1/genética , Sirtuína 1/metabolismo , Traumatismos da Medula Espinal/tratamento farmacológico , Traumatismos da Medula Espinal/genética , Traumatismos da Medula Espinal/metabolismo , Proteína 1 de Transformação que Contém Domínio 2 de Homologia de Src/genética , Proteína 1 de Transformação que Contém Domínio 2 de Homologia de Src/metabolismoRESUMO
Ligamentum flavum (LF) hypertrophy (LFH) has been recognised as one of the key contributors to lumbar spinal stenosis. Currently, no effective methods are available to ameliorate this hypertrophy. In this study, human umbilical cord mesenchymal stromal cell-derived extracellular vesicles (hUCMSC-EVs) were introduced for the first time as promising vehicles for drug delivery to treat LFH. The downregulation of miR-146a-5p and miR-221-3p expressions in human LF tissues negatively correlated with increased LF thickness. The hUCMSC-EVs enriched with these two miRNAs significantly suppressed LFH in vivo and notably ameliorated the progression of transforming growth factor ß1(TGF-ß1)-induced fibrosis in vitro after delivering these two miRNAs to mouse LF cells. The results further demonstrated that miR-146a-5p and miR-221-3p directly bonded to the 3'-UTR regions of SMAD4 mRNA, thereby inhibiting the TGF-ß/SMAD4 signalling pathway. Therefore, this translational study determined the effectiveness of a hUCMSC-EVs-based approach for the treatment of LFH and revealed the critical target of miR-146a-5p and miR-221-3p. Our findings provide new insights into promising therapeutics using a hUCMSC-EVs-based delivery system for patients with lumbar spinal stenosis.
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Osteoblast migration and proliferation are fundamental processes in bone healing. We demonstrated that the G-protein-coupled receptor kinase interacting protein 1(GIT1) is a key regulator of bone mass and osteoblast cell migration, but little is known about GIT1 regulation by upstream signaling systems or the impact of GIT1 on downstream effectors. We found that platelet-derived growth factor (PDGF) stimulated the GIT1 tyrosine phosphorylation in osteoblast cells and increased the association of GIT1 with focal adhesion kinase (FAK) at osteoblast focal adhesions. The Src inhibitor PP2 and FAK siRNA inhibited GIT1 tyrosine phosphorylation and the increased association between GIT1 and FAK following stimulation with PDGF. The spa2 homology domain (SHD) of GIT1 was required for association with FAK. Furthermore, phosphorylation of tyrosine 321 of GIT1, which is localized within the SHD, was critical for association with FAK. Mutagenesis analysis revealed that GIT1Y321F inhibited the increased association between GIT1 and FAK. Immunofluorescent staining revealed that GIT1Y321F inhibited FAK activation in focal adhesions after PGDF stimulation. A cell spreading assay demonstrated that GIT1Y321F also inhibited osteoblast cell motility, while the Boyden chamber assay demonstrated that the GIT1Y321F mutation inhibited PDGF-induced osteoblastic cell migration. Phosphorylation of tyrosine 321 of GIT1 is necessary for PDGF-induced association with FAK, FAK activation in focal adhesions, and for osteoblastic cell migration.
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Proteínas de Ciclo Celular/metabolismo , Movimento Celular , Quinase 1 de Adesão Focal/metabolismo , Adesões Focais/metabolismo , Proteínas Ativadoras de GTPase/metabolismo , Osteoblastos/fisiologia , Fator de Crescimento Derivado de Plaquetas/fisiologia , Animais , Proteínas de Ciclo Celular/genética , Tamanho Celular , Células Cultivadas , Ativação Enzimática , Proteínas Ativadoras de GTPase/genética , Camundongos , Mutagênese Sítio-Dirigida , Osteoblastos/enzimologia , Osteoblastos/metabolismo , Fosforilação , Ligação Proteica , Domínios e Motivos de Interação entre Proteínas , Processamento de Proteína Pós-Traducional , Tirosina/genética , Tirosina/metabolismo , Quinases da Família src/metabolismoRESUMO
Purpose: Both percutaneous endoscopic lumbar discectomy (PELD) and open fenestration discectomy (OFD) are effective and safe surgical procedures for the treatment of LDH. The purpose of this retrospective study was to compare the surgical outcomes of percutaneous endoscopic interlaminar discectomy (PEID) and OFD for single-segment huge lumbar disc herniation (HLDH). Patients and Methods: We retrospectively analyzed 91 patients diagnosed with single-segment HLDH and treated with OFD or PEID. Visual analog scale (VAS), modified Japanese orthopedic association (mJOA) and Oswestry disability index (ODI) were used to assess clinical outcomes at preoperation and postoperatively at 3, 6, 12, and 24months. Modified Macnab criteria were applied to evaluate clinically satisfaction at the final follow-up. Results: In both groups, the VAS and ODI scores at 3, 6, 12, and 24months postoperatively showed a significant decrease and the mJOA score at 3, 6, 12, and 24months postoperatively was significantly increased compared to preoperative results (P<0.001). According to Macnab criteria at the final follow-up, the overall clinically satisfactory rate was 86.67% in the OFD group and 86.96% in the PEID group. There were no significant differences in VAS, ODI, and mJOA scores between the two groups at preoperation and postoperative 3, 6, 12, and 24months, respectively. In the PEID group, the length of hospitalization and the length of incision were significantly shorter than that in the OFD group (P<0.0001). However, there was no significant difference in operative time between groups (P=0.81). Conclusion: Collectively, postoperative clinical results were equally favorable for both procedures, with no statistically significant difference between PEID and OFD at the two-year of follow-up. No serious complication was observed in two groups. Compared with the traditional surgery, PEID has the following benefits: less trauma, less bleeding, speedy recovery, and shorter hospitalization. Therefore, PEID may be a promising alternative to traditional surgery.
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As an important hormone that regulates the balance of calcium and phosphorus, parathyroid hormone (PTH) has also been found to have an important function in intervertebral disc degeneration (IVDD). Our aim was to investigate the mechanism by which PTH alleviates IVDD. In this study, the PTH 1 receptor was found to be highly expressed in severely degenerated human nucleus pulposus (NP) cells. We found in the mouse model of IVDD that supplementation with exogenous PTH alleviated the narrowing of the intervertebral space and the degradation of the extracellular matrix (ECM) caused by tail suspension (TS). In addition, inflammation, oxidative stress, and apoptosis levels were significantly increased in the intervertebral disc tissues of TS-induced mice, and the activity of NP cells was decreased. TS also led to the downregulation of Sonic hedgehog (SHH) signalling pathway-related signal molecules in NP cells such as SHH, Smoothened, and GLI1. However, supplementation with exogenous PTH can reverse these changes. In vitro, PTH also promotes the activity of NP cells and the secretion of ECM. However, the antagonist of the SHH signalling pathway can inhibit the therapeutic effect of PTH on NP cells. In addition, a cAMP-response element-binding protein, as an important transcription factor, was found to mediate the promotion of PTH on the SHH signalling pathway. Our results revealed that PTH can alleviate IVDD by inhibiting inflammation, oxidative stress, and apoptosis and improving the activity of NP cells via activating the SHH signalling pathway.
Assuntos
Proteína de Ligação a CREB/metabolismo , Proteínas Hedgehog/metabolismo , Inflamação/tratamento farmacológico , Degeneração do Disco Intervertebral/tratamento farmacológico , Hormônio Paratireóideo/uso terapêutico , Animais , Apoptose , Modelos Animais de Doenças , Humanos , Masculino , Camundongos , Estresse Oxidativo , Hormônio Paratireóideo/farmacologia , Transdução de SinaisRESUMO
Intervertebral disk degeneration (IDD) is a major cause of low back pain that becomes a prevalent age-related disease. However, the pathophysiological processes behind IDD are rarely known. Here, we used bioinformatics analysis based on the microarray datasets (GSE34095) to identify the differentially expressed genes (DEGs) as biomarkers and therapeutic targets in degenerated discs. From the previous studies, oxidative stress has been notified as a positive inducement of IDD, which causes DNA damage and accelerates cell senescence. Polyamine oxidase (PAOX), a member of the observed 1057 DEGs, is involved in polyamine metabolism and influences the oxidative balance in cells. However, it is uncertain if the IDD is implicated in the dysregulation of PAOX and polyamine metabolism. This study firstly verified the PAOX upregulation in human degenerated disc samples and applied an IL-1ß-induced nucleus pulposus (NP) cell degeneration model to demonstrate that spermidine supplementation balanced polyamine metabolism and delayed NP cell senescence. Moreover, we confirmed that spermidine/N-acetylcysteine supplementation or Cdkn2a gene deletion stabilized the polyamine metabolism, suppressed oxidative stress, and therefore delayed the progress of IDD in older mice. Collectively, our study highlights the role of polyamine metabolism in IDD and foresees spermidine would be the advanced therapeutical drug for IDD.
Assuntos
Senescência Celular/fisiologia , Biologia Computacional/métodos , Degeneração do Disco Intervertebral/terapia , Núcleo Pulposo/metabolismo , Poliaminas/metabolismo , Animais , Células Cultivadas , Humanos , Degeneração do Disco Intervertebral/fisiopatologia , Camundongos , Estresse OxidativoRESUMO
Spinal cord injury (SCI) is a severe traumatic condition. The loss of the bundle of axons involved in motor conduction in the spinal cord after SCI is the main cause of motor function injury. Presently, axon regeneration in the spinal cord has been studied extensively, but it remains unclear how axon growth is regulated in an inflammatory environment at the cellular level. In the present study, GIT1 knockout (KO) mouse neurons were cultured in a microfluidic device to simulate the growth of axons in an inflammatory environment. The molecular regulation of axon growth in an inflammatory environment by GIT1 was then investigated. We found that the axon growth of GIT1 KO mouse neurons was restricted in an inflammatory environment. Further investigations revealed that in both axons and cell bodies in the inflammatory environment, GIT1 phosphorylated ERK, promoted the entry of Nrf2 into the nucleus, and promoted the transcription of MAP1B, thereby increasing the levels of MAP1B and p-MAP1B and promoting axon growth. We also found that MAP1B could be translated locally in axons and transported in cell bodies and axons. In conclusion, we found that GIT1 regulated axon growth in an inflammatory environment. This provided a theoretical basis for axon regeneration in an inflammatory environment after SCI to develop new treatment options for axon regeneration.
Assuntos
Axônios , Regeneração Nervosa , Animais , Axônios/fisiologia , Proteínas de Ciclo Celular/metabolismo , Proteínas Ativadoras de GTPase/metabolismo , Camundongos , Proteínas Associadas aos Microtúbulos/metabolismo , FosforilaçãoRESUMO
OBJECTIVE: To evaluate the curative efficacy by comparing perioperative characteristics and 1.5-year observational outcomes in 1-segment lumbar spondylolisthesis between traditional minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF) and optimized Endoscopic TLIF techniques. METHODS: The study was a single-center, randomized controlled trial comparing two different treatment approaches for 1-segment lumbar spondylolisthesis. 102 patients treated by MIS-TLIF (48 cases) or Endo-TLIF (54 cases) were included from March 2018 to April 2019. Perioperative parameters and clinical outcomes were evaluated. Degree of slip were measured, and fusion rates were determined at 18 months after surgery. RESULTS: The Endo-TLIF group had similar return to work time and rate. Blood loss, left bed time, analgesic ratio were significantly less in Endo-TLIF group. The Endo-TLIF group had a significantly longer operative time. Significant postoperative reduction in %slip was showed in both groups. The VAS and ODI improved significantly in both groups after surgery. Significant decreases in low-back pain in Endo-TLIF group were found at postoperative day 1 and 3 months. The fusion rate in the two groups was similar. CONCLUSION: Endo-TLIF surgery with a C-shaped working tube and a visualization system may be regarded as an efficient alternative surgery for 1-segment lumbar spondylolisthesis. It is a safe and minimally invasive way to perform this surgery and has shown satisfactory clinical outcomes. TRIAL REGISTRATION: ChiCTR1800015197, 13 March 2018. TRIAL REGISTRY: Chinese Clinical Trial Registry. Registered 13 March 2018. http://www.chictr.org.cn/showproj.aspx?proj=25865.
Assuntos
Artroscopia , Vértebras Lombares/cirurgia , Avaliação de Resultados em Cuidados de Saúde , Fusão Vertebral , Espondilolistese/cirurgia , Idoso , Artroscopia/métodos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Estudos Prospectivos , Fusão Vertebral/instrumentação , Fusão Vertebral/métodosRESUMO
Robot-assisted orthopedic surgery has great application prospects, and the accuracy of the robot is the key to its overall performance. The aim of this study was to develop a new orthopedic surgical robot to assist in spinal surgeries and to compare its feasibility and accuracy with the existing orthopedic robot. A new type of high-precision orthopedic surgical robot (Tuoshou) was developed. A multicenter, randomized controlled trial was carried out to compare the Tuoshou with the TiRobot (TINAVI Medical Technologies Co., Ltd., Beijing) to evaluate the accuracy and safety of their navigation and positioning. A total of 112 patients were randomized, and 108 patients completed the study. The position deviation of the Kirschner wire placement in the Tuoshou group was smaller than that in the TiRobot group (p = 0.014). The Tuoshou group was better than the TiRobot group in terms of the pedicle screw insertion accuracy (p = 0.016) and entry point deviation (p < 0.001). No differences were observed in endpoint deviation (p = 0.170), axial deviation (p = 0.170), sagittal deviation (p = 0.324), and spatial deviation (p = 0.299). There was no difference in security indicators. The new orthopedic surgical robot was highly accurate and optimized for clinical practice, making it suitable for clinical application.