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A divergent synthesis of podophyllotoxin derivatives from simple and readily available starting materials through a late-stage functionalization strategy by rhodium catalysis is reported here. This strategy uses the ketone and oxime in substrates as directing groups. Four kinds of novel podophyllotoxin derivatives have been obtained without any erosion of the enantiopurity, thus indicating the broad substrate scope of this method. Additionally, by using the newly developed strategy, 9 aa, which exhibited excellent anticancer activity, can be prepared by a sequential transformation. In particularly, 9 aa suppressed HeLa cells with IC50 values of 74.5â nM, thus providing a promising lead compound for future drug discovery.
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Invited for the cover of this issue are Chunpu Li, Hong Liu and co-workers at Shanghai Institute of Materia Medica, Nanjing University of Chinese Medicine, and Hangzhou Institute for Advanced Study. The image depicts rhodium catalysis converting the readily available podophyllotoxin into four kinds of novel derivatives. Read the full text of the article at 10.1002/chem.202300960.
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Angiogenesis, formation of new blood vessels from the existing vascular network, is a hallmark of cancer cells that leads to tumor vascular proliferation and metastasis. This process is mediated through the binding interaction of VEGF-A with VEGF receptors. However, the balance between pro-angiogenic and anti-angiogenic effect after ligand binding yet remains elusive and is therefore challenging to manipulate. To interrogate this interaction, herein we designed a few sulfono-γ-AA peptide based helical peptidomimetics that could effectively mimic a key binding interface on VEGF (helix-α1) for VEGFR recognition. Intriguingly, although both sulfono-γ-AA peptide sequences V2 and V3 bound to VEGF receptors tightly, in vitro angiogenesis assays demonstrated that V3 potently inhibited angiogenesis, whereas V2 activated angiogenesis effectively instead. Our findings suggested that this distinct modulation of angiogenesis might be due to the result of selective binding of V2 to VEGFR-1 and V3 to VEGFR-2, respectively. These molecules thus provide us a key to switch the angiogenic signaling, a biological process that balances the effects of pro-angiogenic and anti-angiogenic factors, where imbalances lead to several diseases including cancer. In addition, both V2 and V3 exhibited remarkable stability toward proteolytic hydrolysis, suggesting that V2 and V3 are promising therapeutic agents for the intervention of disease conditions arising due to angiogenic imbalances and could also be used as novel molecular switching probes to interrogate the mechanism of VEGFR signaling. The findings also further demonstrated the potential of sulfono-γ-AA peptides to mimic the α-helical domain for protein recognition and modulation of protein-protein interactions.
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Factor A de Crecimiento Endotelial VascularRESUMEN
Papillary thyroid cancer (PTC) is a common malignancy. MicroRNAs (miRNAs) may act as oncogenes or tumor suppressor genes. However, the role of miR-451a in PTC is not fully understood. Hence, the objective of the study was to research the effect and mechanism of miR-451a in PTC. Differentially expressed miRNAs between GSE113629 and GSE103996 databases were assessed by Venn diagram. miR-451a and its downstream target genes were assessed by RT-PCR and Western blot. The proliferation, invasion, and apoptosis were determined by CCK-8, EdU, transwell, and flow cytometry assays. Dual-luciferase reporter assay were used to evaluated the target of miR-451a. Xenografted tumors was used to explore the function of miR-451a in vivo. Pathological changes and related protein expression were measured by HE staining and immunohistochemistry. MiR-451a was downregulated in PTC tissues and blood, and low expression of miR-451a was related to short overall survival, serious lymph node metastasis and high TNM grade in PTC patients. Moreover, increase of miR-451a restrained the proliferation and invasion and accelerated the apoptosis. Furthermore, miR-451a repressed VEGF signaling pathway. Importantly, miR-451a was demonstrated to target DCBLD2 and AKT1. Overexpression of DCBLD2 and AKT1 could restore the effect of miR-451a on PTC cells. In addition, miR-451a reduced the growth of xenografted tumors in vivo. The data suggested that miR-451a attenuated the proliferation, invasion and promoted apoptosis in PTC cells via inhibiting DCBLD2 and AKT1.
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Carcinoma Papilar , MicroARNs , Neoplasias de la Tiroides , Humanos , Apoptosis/genética , Carcinoma Papilar/genética , Carcinoma Papilar/metabolismo , Carcinoma Papilar/patología , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Regulación Neoplásica de la Expresión Génica/genética , Proteínas de la Membrana/genética , MicroARNs/genética , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Cáncer Papilar Tiroideo/genética , Cáncer Papilar Tiroideo/patología , Neoplasias de la Tiroides/genética , Neoplasias de la Tiroides/metabolismoRESUMEN
BACKGROUND: Acute myeloid leukemia (AML) is a molecularly heterogeneous disease that accounts for approximately 25% of childhood leukemia cases. In this study, we aimed to identify survival-associated genes in pediatric AML patients and investigate potential immunotherapy targets. METHODS: After retrieving and processing the data from Gene Expression Omnibus (GEO) web resource, we determined hub genes in AML. Bioinformatics technology was applied to identify key genes and perform functional analysis. Finally, we investigated the correlation between the key gene and the infiltration levels of tumor-infiltrating immune cells. RESULTS: High protein tyrosine phosphatase receptor-type C (PTPRC) expression was associated with worse overall survival rate (p < 0.001) in 287 pediatric AML patients. The results of risk subgroup analyses were similar in the high-risk and low-risk groups (p = 0.007; p = 0.013). Meanwhile, high expression of PTPRC was an independent adverse prognostic factor for overall survival (p = 0.04). Moreover, the results of immune infiltration assessment demonstrated that the expression level of PTPRC was significantly correlated with the infiltration level of activated dendritic cells (p < 0.001). CONCLUSIONS: Overexpression of PTPRC indicates poor prognosis, and its expression level is correlated with the infiltration level of activated dendritic cells. PTPRC could be a promising immunotherapy target for pediatric AML.
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Leucemia Mieloide Aguda , Monoéster Fosfórico Hidrolasas , Niño , Biología Computacional , Humanos , Leucemia Mieloide Aguda/diagnóstico , Leucemia Mieloide Aguda/genética , Antígenos Comunes de Leucocito , Pronóstico , Tasa de SupervivenciaRESUMEN
The rational design of α-helix-mimicking peptidomimetics provides a streamlined approach to discover potent inhibitors for protein-protein interactions (PPIs). However, designing cell-penetrating long peptidomimetic scaffolds equipped with various functional groups necessary for interacting with large protein-binding interfaces remains challenging. This is particularly true for targeting ß-catenin/BCL9 PPIs. Here we designed a series of unprecedented helical sulfono-γ-AApeptides that mimic the binding mode of the α-helical HD2 domain of B Cell Lymphoma 9 (BCL9). Our studies show that sulfono-γ-AApeptides can structurally and functionally mimic the α-helical domain of BCL9 and selectively disrupt ß-catenin/BCL9 PPIs with even higher potency. More intriguingly, these sulfono-γ-AApeptides can enter cancer cells, bind with ß-catenin and disrupt ß-catenin/BCL9 PPIs, and exhibit excellent cellular activity, which is much more potent than the BCL9 peptide. Furthermore, our enzymatic stability studies demonstrate the remarkable stability of the helical sulfono-γ-AApeptides, with no degradation in the presence of pronase for 24 h, augmenting their biological potential. This work represents not only an example of helical sulfono-γ-AApeptides that mimic α-helix and disrupt protein-protein interactions, but also an excellent example of potent, selective, and cell-permeable unnatural foldameric peptidomimetics that disrupt the ß-catenin/BCL9 PPI. The design of helical sulfono-γ-AApeptides may lead to a new strategy to modulate a myriad of protein-protein interactions.
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Péptidos , Unión Proteica/efectos de los fármacos , Conformación Proteica en Hélice alfa , Factores de Transcripción/metabolismo , beta Catenina/metabolismo , Línea Celular Tumoral , Humanos , Péptidos/química , Péptidos/metabolismo , Péptidos/farmacología , Peptidomiméticos , Mapas de Interacción de Proteínas/efectos de los fármacos , Factores de Transcripción/antagonistas & inhibidores , beta Catenina/antagonistas & inhibidoresRESUMEN
Mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1) is a key adaptor protein that regulates the NF-κB pathway, in which MALT1 functions as a scaffold protein and protease to trigger downstream signals. The abnormal expression of MALT1 is closely associated with lymphomagenesis and other diseases, including solid tumors and autoimmune diseases. MALT1 is the only protease in the underlying pathogenesis of these diseases, and its proteolytic activity can be pharmacologically regulated. Therefore, MALT1 is a potential and promising target for anti-lymphoma and other MALT1-related disease treatments. Currently, the development of MALT1 inhibitors is still in its early stages. This review presents an overview of MALT1, particularly its X-ray structures and biological functions, and elaborates on the pathogenesis of diseases associated with its dysregulation. We then summarize previously reported MALT1 inhibitors, focusing on their molecular structure, biological activity, structure-activity relationship, and limitations. Finally, we propose future research directions to accelerate the discovery of novel MALT1 inhibitors with clinical applications. Overall, this review provides a comprehensive and systematic overview of MALT1-related research advances and serves as a theoretical basis for drug discovery and research.
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Enfermedades Autoinmunes , Linfoma , Humanos , Proteína 1 de la Translocación del Linfoma del Tejido Linfático Asociado a Mucosas , FN-kappa B , Proteínas de Neoplasias/genéticaRESUMEN
A facile access to highly fused tetracyclic indeno-1,2-benzothiazines has been established via a Rh(III)-catalyzed C-H bond activation and intramolecular annulation cascade between sulfoximides and all-carbon diazo indandiones. This strategy is characterized by the fact that the diazo coupling partners do not require preactivation, along with its high efficiency, broad substrate generality, and facile transformation. Particularly, the highly conjugated tetracyclic products demonstrate good optical properties and can easily enter cells to emit bright fluorescence for live cell imaging.
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Rodio , Carbono , CatálisisRESUMEN
BACKGROUND: Severe neurotoxicity after chimeric antigen receptor T cell (CAR-T) therapy can be a crucial lifethreatening event in diffuse large B-cell lymphoma (DLBCL), and management of those toxicities is still a serious clinical challenge. The underlying mechanisms of CAR-T cell-mediated neurotoxicity remain poorly elucidated because very few studies examine the intact tumor microenvironment before CAR-T cell infusion. Herein, we pur-posed to identify differentially expressed genes (DEGs) related to CAR-T cell-mediated neurotoxicity in the DLBCL microenvironment before CAR-T cell infusion and reveal their potential mechanisms. METHODS: The mRNA expression profile data of GSE153438 were obtained from the GEO database. The GSE153438 dataset includes 26 samples with non-severe neurotoxicity (grade 0 - 2) and 10 samples with severe neurotoxicity (grade 3 or higher). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) patway enrichment assessment was carried out. We screened the hub gene by protein-protein interaction (PPI) network analysis and Cytoscape software. Gene set enrichment analysis (GSEA) was also analyzed with the GSEA software. Moreover, the predictive value of the hub gene for severe neurotoxicity was evaluated via receiver operating characteristic (ROC) curve analysis. RESULTS: We identified a total of 25 up-regulated DEGs and 26 downregulated DEGs associated with CAR-T cell-mediated neurotoxicity in the DLBCL microenvironment before CAR-T cell infusion. Results of GO analysis showed that DEGs were mainly enriched in T cell activation, leukocyte cell-cell adhesion, and positive regulation of cell adhesion. The KEGG analysis revealed that DEGs were significantly enriched in T cell receptor signaling pathway, cell adhesion molecules, and Epstein-Barr virus infection. GSEA revealed that the glycolysis pathway was significantly associated with severe neurotoxicity. The top centrality hub gene GZMB was identified from the PPI network. ROC curve analysis showed that GZMB had a potential predictive value for severe neurotoxicity. CONCLUSIONS: In DLBCL microenvironment before CAR-T cell infusion, we identified T cell activation and glycolysis pathways significantly associated with CAR-T cell-mediated severe neurotoxicity. GZMB might be used as a predictive and therapeutic molecular marker for neurotoxicity. The study suggested that the tumor microenviron-ment before CAR-T cell infusion plays an essential role in the early prediction of neurotoxicity.
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Infecciones por Virus de Epstein-Barr , Linfoma de Células B Grandes Difuso , Biología Computacional , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Redes Reguladoras de Genes , Herpesvirus Humano 4 , Humanos , Linfoma de Células B Grandes Difuso/genética , Transcriptoma , Microambiente Tumoral/genéticaRESUMEN
A catalytic system-controlled divergent reaction strategy was here reported to construct four types of intriguing spiroheterocyclic skeletons from simple and readily available starting materials via a precise chemical bond activation/[n+1] annulation cascade. The tetraazaspiroheterocyclic and trizazspiroheterocyclic scaffolds could be independently constructed by a selective N-N bond activation/[n+1] annulation cascade, a C(sp2 )-H activation/[4+1] annulation and a novel tandem C(sp2 )-H/C(sp3 )-H bond activation/[4+1] annulation strategy, along with a broad scope of substrates, moderate to excellent yields and valuable transformations. More importantly, in these transformations, we are the first time to capture a N-N bond activation and a C(sp3 )-H bond activation of pyrazolidinones under different catalytic system.
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A facile access to the polycyclic-fused pyrano[2,3-b]pyridines has been established under room temperature via Rh(III)-catalyzed C-H bond activation and intramolecular cascade annulation. This strategy features high efficiency, unique versatility, and generality and it can occur under mild conditions in good to excellent yields. More importantly, this strategy can be extended to the late-stage functionalization of drugs possessing the CN group.
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BACKGROUND: Osteosarcoma is the most frequent primary malignant tumor of bone. SLC19A1 has been explored as a novel biomarker in some cancers. In this research, the diagnostic and prognostic value of SLC19A1 expression in osteosarcoma was evaluated by bioinformatics analysis. Data were sourced from the Gene Expression Omnibus (GEO) database. METHODS: Gene expression data and clinical materials of patients with osteosarcoma were collected from GSE42352 and GSE21257 datasets. The mRNA expression of SLC19A1 was compared between osteosarcoma cells and mesenchyme stem cells with the Wilcoxon rank-sum test. Moreover, receiver operating characteristic (ROC) curve analysis was performed to determine the diagnostic merit of SLC19A1 for osteosarcoma. The relationship between SLC19A1 and clinicopathological characteristics was analyzed using logistic regression. Besides, the correlation between SLC19A1 and survival rate was assessed using Kaplan-Meier and Cox regression. The biological functions of SLC19A1 were annotated and evaluated through gene set enrichment analysis (GSEA) and gene set variation analysis (GSVA). RESULTS: SLC19A1 was significantly highly expressed in osteosarcoma cells (p < 0.001). The ROC curve showed an area under the curve of 0.899, which indicated a high diagnostic value. High SLC19A1 expression showed a negative correlation with Huvos grade [odds ratio (OR) = 0.09 for III vs. I, p = 0.014]. Kaplan-Meier survival analysis showed that the overall survival (OS) of the patients with high SLC19A1 expression was significantly poorer than the low SLC19A1 expression group (p = 0.016). The univariate analysis revealed that high SLC19A1 expression was associated with poor OS [p = 0.013, hazard ratio (HR) = 6.74, 95% CI = 1.49 - 30.46]. The multivariate analysis revealed that SLC19A1 expression (p = 0.014, HR = 8.03, 95% CI = 1.52 - 42.51) was independently correlated with OS. GSEA showed that genes in high expression group of SLC19A1 were enriched in KEGG pathways, including "Glyoxylate and dicarboxylate metabolism", "Oxidative phosphorylation", "Aminoacyl tRNA biosynthesis", "Base excision repair", "Pyrimidine metabolism" and "Proteasome". GSVA further suggested their importance in the progression of osteosarcoma. CONCLUSIONS: SLC19A1 may be a potential biomarker for diagnosis and prognosis in osteosarcoma.
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Neoplasias Óseas , Osteosarcoma , Biomarcadores de Tumor/genética , Neoplasias Óseas/diagnóstico , Neoplasias Óseas/genética , Humanos , Estimación de Kaplan-Meier , Osteosarcoma/diagnóstico , Osteosarcoma/genética , Pronóstico , Proteína Portadora de Folato ReducidoRESUMEN
OBJECTIVE: To reveal the molecular mechanism underlying the pathogenesis of HCM and find new effective therapeutic strategies using a systematic biological approach. METHODS: The WGCNA algorithm was applied to building the co-expression network of HCM samples. A sample cluster analysis was performed using the hclust tool and a co-expression module was constructed. The WGCNA algorithm was used to study the interactive connection between co-expression modules and draw a heat map to show the strength of interactions between modules. The genetic information of the respective modules was mapped to the associated GO terms and KEGG pathways, and the Hub Genes with the highest connectivity in each module were identified. The Wilcoxon test was used to verify the expression level of hub genes between HCM and normal samples, and the "pROC" R package was used to verify the possibility of hub genes as biomarkers. Finally, the potential functions of hub genes were analyzed by GSEA software. RESULTS: Seven co-expression modules were constructed using sample clustering analysis. GO and KEGG enrichment analysis judged that the turquoise module is an important module. The hub genes of each module are RPL35A for module Black, FH for module Blue, PREI3 for module Brown, CREB1 for module Green, LOC641848 for module Pink, MYH7 for module Turquoise and MYL6 for module Yellow. The results of the differential expression analysis indicate that MYH7 and FH are considered true hub genes. In addition, the ROC curves revealed their high diagnostic value as biomarkers for HCM. Finally, in the results of the GSEA analysis, MYH7 and FH highly expressed genes were enriched with the "proteasome" and a "PPAR signaling pathway," respectively. CONCLUSIONS: The MYH7 and FH genes may be the true hub genes of HCM. Their respective enriched pathways, namely the "proteasome" and the "PPAR signaling pathway," may play an important role in the development of HCM.
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Biomarcadores , Cardiomiopatía Hipertrófica/diagnóstico , Cardiomiopatía Hipertrófica/genética , Perfilación de la Expresión Génica , Redes Reguladoras de Genes , Biología Computacional/métodos , Bases de Datos Genéticas , Regulación de la Expresión Génica , Ontología de Genes , Humanos , Pronóstico , TranscriptomaRESUMEN
A mild and facile Cp*Rh(III)-catalyzed C-H activation and intramolecular cascade annulation protocol has been proposed for the furnishing of highly fused isochromeno-1,2-benzothiazines scaffolds using S-phenylsulfoximides and 4-diazoisochroman-3-imine as substrates under room temperature. This method features diverse substituents and functional groups tolerance and relatively mild reaction conditions with moderate to excellent yields. Additionally, retentive configuration of sulfoximides in the conversion has been verified.
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Sulfóxidos/química , Tiazinas/química , Estructura Molecular , Rodio/química , Estereoisomerismo , TemperaturaRESUMEN
Sirtuins (SIRT1-7) are a family of NAD+-dependent deacetylases. They regulate many physiological processes and play important roles in inflammation, diabetes, cancers, and neurodegeneration diseases. Sirtuin inhibitors have potential applications in the treatment of neurodegenerative diseases and various cancers. Herein, we identified new sirtuin inhibitors based on the scaffold of 8-mercapto-3,7-dihydro-1H-purine-2,6-dione. To elucidate the inhibitory mechanism, the binding modes of the inhibitors in SIRT3 were established by molecular docking, showing that the inhibitors occupy the acetyl lysine binding site and interact with SIRT3, mainly through hydrophobic interactions. The interactions were validated by site-directed mutagenesis of SIRT3 and structure-activity relationship analysis of the inhibitors. Consistently, enzyme kinetic assays and microscale thermophoresis showed that these compounds are competitive inhibitors to the acetyl substrate, and mix-type inhibitors to NAD+. Furthermore, we demonstrated that the compounds are potent SIRT1/2/3/5 pan-inhibitors. This study provides novel hits for developing more potent sirtuin inhibitors.
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Sirtuinas/antagonistas & inhibidores , Sirtuinas/metabolismo , Teofilina/química , Teofilina/farmacología , Sitios de Unión , Diseño de Fármacos , Humanos , Cinética , Simulación del Acoplamiento Molecular , Estructura Molecular , Sirtuina 1/antagonistas & inhibidores , Sirtuina 1/metabolismo , Sirtuina 2/antagonistas & inhibidores , Sirtuina 3/antagonistas & inhibidores , Sirtuina 3/metabolismo , Relación Estructura-ActividadRESUMEN
Unprecedented Rh(III)-catalyzed C-H bond difluorovinylation of indoles has been successfully developed, and this method provided an example of direct difluorovinylation reaction through C-H bond activation which was rarely reported. In this context, N-ethoxycarbamoyl served as the directing group and 2,2-difluorovinyl tosylates were used for the construction of difluorovinyl-substituted indoles. This method provided a practical strategy for difluorovinylation of indoles with moderate to good yields and is characterized by the broad synthetic utility, mild conditions, and high efficiency.
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Bencenosulfonatos/química , Hidrocarburos Fluorados/síntesis química , Indoles/química , Compuestos Organometálicos/química , Rodio/química , Catálisis , Hidrocarburos Fluorados/química , Estructura MolecularRESUMEN
An efficient approach to trans-indolylvinylboronate derivatives has been developed using an Rh-catalyzed hydroarylation of alkyne N-methyliminodiacetic acid boronates via C-H activation. This protocol constitutes a straightforward route for the synthesis of B-containing aza-heterocycles in good yields with excellent functional group tolerance.
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There is an urgent need to develop effective therapies for ischemic stroke, but the complicated pathological processes after ischemia make doing so difficult. In the current study, we identified a novel diaryl acylhydrazone derivative, A11, which has multiple neuroprotective properties in ischemic stroke models. First, A11 was demonstrated to induce neuroprotection against ischemic injury in a dose-dependent manner (from 0.3 to 3 µM) in three in vitro experimental ischemic stroke models: oxygen glucose deprivation (OGD), hydrogen peroxide, and glutamate-stimulated neuronal cell injury models. Moreover, A11 was able to potently alleviate three critical pathological changes, apoptosis, oxidative stress, and mitochondrial dysfunction, following ischemic insult in neuronal cells. Further analysis revealed that A11 upregulated the phosphorylation levels of protein kinase B (AKT) and extracellular signal-regulated kinase (ERK) in OGD-exposed neuronal cells, suggesting joint activation of the phosphoinositide 3-kinase (PI3K)/AKT and mitogen-activated protein kinase (MEK)/ERK pathways. In rats with middle cerebral artery occlusion, single-dose administration of A11 (3 mg/kg per day, i.v.) at the onset of reperfusion significantly reduced the infarct volumes and ameliorated neurological deficits. Our study, for the first time, reports the anti-ischemic effect of diaryl acylhydrazone chemical entities, especially A11, which acts on multiple ischemia-associated pathological processes. Our results may provide new clues for the development of an effective therapeutic agent for ischemic stroke.
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Hidrazonas/uso terapéutico , Infarto de la Arteria Cerebral Media/tratamiento farmacológico , Fármacos Neuroprotectores/uso terapéutico , Daño por Reperfusión/tratamiento farmacológico , Animales , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Humanos , Hidrazonas/farmacología , Masculino , Enfermedades Mitocondriales/tratamiento farmacológico , Neuronas/efectos de los fármacos , Fármacos Neuroprotectores/farmacología , Estrés Oxidativo/efectos de los fármacos , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: Granulocyte colony-stimulating factor (G-CSF) is the critical regulator of the proliferation, differentiation, and survival of granulocytes. Recently, it has been shown that G-CSF can adversely affect bone health in both animal models and patients. Here, the authors aimed to investigate whether G-CSF could inhibit the growth of osteoblasts and osteocytes by regulating nitric oxide. METHODS: The C57BL/6 mice were divided into the control group, G-CSF treatment group and recovery group (G-CSF+L-NAME). The morphology of femurs was assessed by histology and immunohistochemistry. The expression of apoptosis-related molecules in femurs was detected by immunohistochemistry and quantitative RT-PCR, respectively. To examine if neutrophil-secreted factors can induce apoptosis in osteoblasts, Gr1-positive (Gr1+) neutrophils from the bone marrow of wild-type mice were sorted and co-cultured with MC3T3 pre-osteoblasts for 2 days. RESULTS: The number of osteoblasts and newly embedding osteocytes significantly decreased and markers related to osteoblasts and osteocytes were downregulated in the G-CSF treatment compared to the control group. Moreover, G-CSF treatment did not change proliferation markers but induced apoptosis in osteoblast-lineage cells. The combined treatment of mice with G-CSF and a nitric oxide inhibitor partially restored the number of osteoblasts and osteocyte parameters. CONCLUSIONS: The G-CSF can inhibit osteoblasts and osteocytes by upregulating nitric oxide.
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Factor Estimulante de Colonias de Granulocitos/farmacología , Neutrófilos/efectos de los fármacos , Óxido Nítrico/biosíntesis , Osteoblastos/efectos de los fármacos , Osteocitos/efectos de los fármacos , Animales , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Femenino , Ratones , Ratones Endogámicos C57BL , Neutrófilos/citología , Neutrófilos/metabolismo , Osteoblastos/citología , Osteocitos/citología , Osteocitos/metabolismo , Regulación hacia Arriba/efectos de los fármacosRESUMEN
BACKGROUND: Dyslipidaemia is a well-known risk factor for the development of atherosclerosis, however, little is known about the effect of dyslipidaemia on intervertebral disc degeneration (IVDD). Thus, the purpose of this study is to investigate the relationship between dyslipidaemia and IVDD, and to identify the possible mechanism by which dyslipidaemia aggravates the degeneration of intervertebral discs. METHODS: Hyperlipidaemia rats were induced, thirty male Wistar rats were randomly divided into two groups: normal chow diet control group (CON) and high-fat diet group (HFD) for 8 weeks. And then, a rat disc degeneration model was established, rats were divided into three experimental groups: the normal chow diet + sham surgery group (CON-Sham); the normal chow diet + needle puncture group (CON-NP); and the high-fat diet + needle puncture group (HFD-NP), all rats were continually fed with normal chow diet or HFD 8 weeks. At the end of the experiment, the discs were harvested and histomorphological analysis, immunohistochemistry staining, real-time PCR and western blot were performed for all groups. RESULTS: The degenerative histological score of disc in the HFD-NP group was significantly higher than the CON-NP group. Immunohistochemical analysis revealed remarkable reductions in aggrecan and collagen type II expressions, and significant increases in IL-1ß, TNF-α, MMP-13, HIF-1α and P65 expression in the HFD-NP group. RT-PCR and western blot analysis showed that the mRNA levels and protein expressions of MMP-13 and TIMP-1 were higher in the HFD-NP group. CONCLUSIONS: Hyperlipidaemia resulted in an exaggerated degenerative changes and altered expression and transcription of the degeneration-associated molecules in the rat disc tissue. These results raise the possibility that hyperlipidaemia may accelerate the progression of disc degeneration.