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1.
Gene ; 715: 144029, 2019 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-31376409

RESUMO

Intervertebral disc degeneration (IDD) is a major cause of lower back pain, but the specific molecular mechanisms governing its development are poorly characterized. This study sought to assess to what extent HOTAIR, a long non-coding (Lnc) RNA is expressed in IDD and regulates the apoptotic death of nucleus pulposus (NP) cells. We therefore used real-time qPCR to measure HOTAIR and microRNA(miR)-34a-5p in degenerative NP cells, and then validated their functional relevance via overexpressing them in these NP cells. We further verified the targets of these RNA constructs in 293 T cells through the use of a dual luciferase reporter assay. We further measured NP cell apoptosis via flow cytometry and Notch1 expression via western blotting. Our results indicated that IDD was linked with decreased HOTAIR expression relative to regular NP cells, and overexpressing this lncRNA was linked to reduced apoptotic NP cell death, whereas overexpressing miR-34a-5p had the opposite effect. We found that HOTAIR served as a miR-34a-5p sponge, sequestering this miRNA and thereby down regulating genes linked to apoptosis through the Notch signaling pathway. Even in naturally degenerated NP cells, HOTAIR delayed the onset of apoptosis. Together these results reveal that a HOTAIR/miR-34a-5p/Notch1 signaling pathway may regulate the development of IDD, potentially making HOTAIR a viable target for treatment of this disease.


Assuntos
Apoptose , Regulação para Baixo , Degeneração do Disco Intervertebral/metabolismo , MicroRNAs/biossíntese , RNA Longo não Codificante/biossíntese , Receptor Notch1/biossíntese , Transdução de Sinais , Adulto , Idoso , Feminino , Humanos , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/patologia , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Núcleo Pulposo/patologia , RNA Longo não Codificante/genética , Receptor Notch1/genética
2.
Life Sci ; 232: 116565, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31251999

RESUMO

HEADINGS AIMS: The present study determined whether nucleus pulposus (NP) cells express hypoxia-inducible factor-2alpha (HIF-2α) and assessed its role in regulating the expression of catabolic factors during intervertebral disc degeneration. MATERIALS AND METHODS: Human degenerated NP tissues were acquired to examine the HIF-2α expression levels using immunohistochemistry, western blotting, and Real-time PCR. Human NP cells were cultivated under normoxic or hypoxic conditions, and the HIF-2α expression was determined. Then, human NP cells were treated with HIF-2α plasmids, HIF-2α siRNA, and tumor necrosis factor-alpha (TNF-α) to evaluate the role of HIF-2α in regulating matrix metalloproteinase (MMP) and aggrecanase expression. An in vivo rabbit disc degeneration model was established to demonstrate that HIF-2α plays a critical role in disc degeneration. KEY FINDINGS: We found that HIF-2α had a markedly elevated expression in human degenerated discs in the Grade III stage. HIF-2α protein and gene transcript levels in vitro were relatively higher under hypoxic conditions. The expression of MMP-13, ADAMTS-4 was decreased significantly in HIF-2α silencing condition, while the over-expression resulted in significantly increased levels of MMP-13 and ADAMTS-4. When cytokine TNF-α was added, HIF-2α was induced by nuclear factor-κB (NF-κB). The in vivo experiments showed that the HIF-2α controlled the catabolic factors MMP-13 and ADAMTS-4 that regulated the collagen II and aggrecan metabolism in disc degeneration. SIGNIFICANCE: HIF-2α is a catabolic regulator in disc degeneration and directly controls the catabolic genes. The suppression of HIF-2α expression leads to decelerates extracellular matrix degradation that might represent a therapeutic target for the degenerative disc.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/biossíntese , Degeneração do Disco Intervertebral/metabolismo , Adulto , Idoso , Agrecanas/metabolismo , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Citocinas/metabolismo , Endopeptidases/metabolismo , Feminino , Humanos , Hipóxia/metabolismo , Disco Intervertebral/metabolismo , Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/patologia , Masculino , Metaloproteinases da Matriz/metabolismo , Pessoa de Meia-Idade , Núcleo Pulposo/metabolismo , Núcleo Pulposo/patologia , Coelhos , Transdução de Sinais/fisiologia , Transcriptoma , Fator de Necrose Tumoral alfa/metabolismo
3.
Medicine (Baltimore) ; 98(21): e15796, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31124977

RESUMO

miRNAs and genes play significant roles in the etiology and pathogenesis of intervertebral disc degeneration (IDD). This study aimed to identify aberrantly expressed miRNAs, genes, and pathways in IDD through a comprehensive bioinformatics analysis.Data of miRNAs expression microarrays (GSE63492) and genes microarrays (GSE23130) were obtained from GEO database. Similarly, aberrantly expressed miRNAs and genes were obtained using GEO2R. In addition, functional and enrichment analyses of selected miRNAs and genes were performed using the DAVID database. Meanwhile, protein-protein interaction (PPI) network was constructed using STRING, and then visualized in Cytoscape.A total of 98 upregulated miRNAs were identified. They were enriched in biological processes of response to organelle, ion binding, cellular nitrogen compound metabolic process, biosynthetic process, small molecule metabolic process, cellular protein modification process, catabolic process, molecular function, neurotrophin TRK receptor signaling pathway, and protein complex. In addition, 1405 high expression protein genes were detected. It indicated enrichment in biological processes, such as translational initiation, nonsense-mediated decay, viral transcription, cell-cell adhesion, rRNA processing, translation, RP-dependent cotranslational protein targeting to membrane, nuclear-transcribed mRNA catabolic process, regulation of mRNA stability, and mRNA splicing via spliceosome and extracellular matrix organization. In addition, pathway analysis exhibited the common enrichment in focal adhesion, Hippo signaling pathway, ECM-receptor interaction, Wnt signaling pathway, PI3K-Akt signaling pathway, endocytosis, proteoglycans in cancer, and so on. The top 10 central genes of PPI network were POTEE, PPP2CA, RPL17, HSP90AA1, POTEF, RPL13A, ACTB, RPL18, RPS24, and HSPA1A.In conclusion, our research proposed abnormally expressed miRNAs, genes, and pathways in IDD through bioinformatics methods, which may provide new insights into the pathogenesis of IDD. Thus, the Hub gene involving POTEE, PPP2CA, RPL17, HSP90AA1, POTEF, RPL13A, ACTB, RPL18, RPS24, and HSPA1A may be biomarkers for accurate diagnosis and treatment of IDD in the future.


Assuntos
Mineração de Dados/métodos , Degeneração do Disco Intervertebral/genética , MicroRNAs/genética , Transdução de Sinais/genética , Revisão Sistemática como Assunto , Biologia Computacional/métodos , Ontologia Genética , Marcadores Genéticos/genética , Humanos , Análise em Microsséries , Mapas de Interação de Proteínas , Projetos de Pesquisa
4.
Int J Mol Sci ; 20(8)2019 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-31027158

RESUMO

Intervertebral disc degeneration (IVDD) is a chronic, expensive, and high-incidence musculoskeletal disorder largely responsible for back/neck and radicular-related pain. It is characterized by progressive degenerative damage of intervertebral tissues along with metabolic alterations of all other vertebral tissues. Despite the high socio-economic impact of IVDD, little is known about its etiology and pathogenesis, and currently, no cure or specific treatments are available. Recent evidence indicates that besides abnormal and excessive mechanical loading, inflammation may be a crucial player in IVDD. Furthermore, obese adipose tissue is characterized by a persistent and low-grade production of systemic pro-inflammatory factors. In this context, chronic low-grade inflammation associated with obesity has been hypothesized as an important contributor to IVDD through different, but still unknown, mechanisms. Adipokines, such as leptin, produced prevalently by white adipose tissues, but also by other cells of mesenchymal origin, particularly cartilage and bone, are cytokine-like hormones involved in important physiologic and pathophysiological processes. Although initially restricted to metabolic functions, adipokines are now viewed as key players of the innate and adaptative immune system and active modulators of the acute and chronic inflammatory response. The goal of this review is to summarize the most recent findings regarding the interrelationships among inflammation, obesity and the pathogenic mechanisms involved in the IVDD, with particular emphasis on the contribution of adipokines and their potential as future therapeutic targets.


Assuntos
Adipocinas/metabolismo , Inflamação/genética , Degeneração do Disco Intervertebral/genética , Obesidade/genética , Humanos , Modelos Biológicos
5.
FEBS Open Bio ; 9(4): 728-735, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30984546

RESUMO

MicroRNAs (miRNAs) are small endogenous non-coding RNAs that can negatively regulate the expression of their complementary mRNA targets, and have been implicated in various pathophysiological processes. In this study, we examined the effect of miR-222-3p on intervertebral disc degeneration (IDD). We found that expression of miR-222-3p was significantly higher in IDD tissues than in normal intervertebral disc tissue, and report that overexpression of miR-222-3p remarkably increased apoptosis and reduced proliferation of nucleus pulposus (NP) cells. In addition, miR-222-3p promoted secretion of matrix metalloproteinase-3, and decreased collagen type II and aggrecan production. Cyclin-dependent kinase inhibitor 1B (CDKN1B) was identified as a direct target of negative regulation by miR-222-3p in NP cells, and expression of miR-222-3p was found to be negatively correlated with that of CDKN1B in IDD tissue. Finally, we observed that transfection with miR-222-3p dramatically reduced CDKN1B expression in NP cells. In conclusion, miR-222-3p may be involved in IDD development, possibly through targeting CDKN1B.


Assuntos
Inibidor de Quinase Dependente de Ciclina p27/genética , Regulação da Expressão Gênica/genética , Degeneração do Disco Intervertebral/genética , MicroRNAs/genética , Apoptose/genética , Proliferação de Células/genética , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Feminino , Humanos , Degeneração do Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/fisiopatologia , Masculino , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Núcleo Pulposo/fisiologia
6.
Cell Mol Biol Lett ; 24: 2, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30936926

RESUMO

Background: MicroRNA (miRNA) plays a vital role in the pathogenesis of intervertebral disc degeneration (IDD). The expression and potential mechanism of miR-573 in human nucleus pulposus (NP) remains to be elucidated. In this study, we aimed to investigate the role of miR-573 in IDD. Methods: Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis was applied to examine the expression of miR-573 and Bax in idiopathic scoliosis tissues and IDD tissues. Human NP cells were employed for analysis. Moreover, the proliferation and apoptosis of NP cells were detected using MTT and flow cytometry assay respectively. The expression levels of Bcl-2, cleaved caspase-3, cleaved caspase-9, caspase-3 and caspase-9 in degenerative NP cells were measured by Western blotting assay. Furthermore, a luciferase reporter assay was used to verify the relationship between miR-573 and Bax. Results: The results revealed that the mRNA expression level of miR-573 was down-regulated whereas Bax was up-regulated notably in degenerative NP cells. In addition, overexpression of miR-573 increased cell viability remarkably, coupled with inhibition of cell apoptosis. The expression level of Bcl-2 was increased while cleaved caspase-3 and cleaved caspase-9 expression levels were decreased in miR-573 overexpression NP cells. Additionally, the bioinformatics analysis underscored that Bax was a direct target gene of miR-573. Conclusion: These results suggest that overexpression of miR-573 inhibited NP cell apoptosis by down-regulating Bax, which proved to be a novel effective strategy for IDD therapies.


Assuntos
Apoptose/genética , Regulação da Expressão Gênica , MicroRNAs/metabolismo , Núcleo Pulposo/metabolismo , Proteína X Associada a bcl-2/metabolismo , Sequência de Bases , Caspase 3/metabolismo , Caspase 9/metabolismo , Proliferação de Células/genética , Sobrevivência Celular/genética , Células Cultivadas , Humanos , Degeneração do Disco Intervertebral/genética , MicroRNAs/genética , Núcleo Pulposo/patologia , Escoliose/genética , Proteína X Associada a bcl-2/genética
7.
Biomed Res Int ; 2019: 9890279, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30915367

RESUMO

Low back pain is a major cause of disability worldwide. Although numerous potential biomarkers for the early diagnosis or treatment of intervertebral disc degeneration (IDD) have been identified subsequent to the development of molecular biology technologies, the mechanisms of IDD remain unknown. Published studies found the unbalance of anabolism and catabolism of annulus fibrosus (AF) played an important role in it. The present study was aimed to identify the potential targets and signaling pathways of IDD, through the combined analysis of differential expression and based on the Gene Expression Omnibus (GEO) dataset from NCBI. PPI Networks Analysis indicated that MMP2 and AGE-RAGE signaling pathway and estrogen signaling pathway may play important roles in initiation and development of IDD. This study forecasted the pathogenesis molecular mechanism of IDD and the potential prognostic and diagnostic biomarkers, but we need to make further molecular biological experiments to confirm our assumptions.


Assuntos
Anel Fibroso/metabolismo , Biomarcadores/metabolismo , Biologia Computacional , Degeneração do Disco Intervertebral/diagnóstico , Anel Fibroso/patologia , Antígenos de Neoplasias/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Humanos , Disco Intervertebral/fisiopatologia , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/patologia , Metaloproteinase 2 da Matriz/genética , Proteínas Quinases Ativadas por Mitógeno/genética , Prognóstico
8.
DNA Cell Biol ; 38(5): 457-467, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30864829

RESUMO

Degenerative disc disease (DDD) is the main cause of low back pain, and the ingrowth of new blood vessels is one of its pathological features. The stromal cell-derived factor 1 (SDF1)/CXCR7 signaling axis plays a role in these physiological and pathological activities. The aims of this study were to explore whether this signaling axis participates in the angiogenesis of degenerated intervertebral discs (IVDs) and to define its underlying mechanism. In this study, we cocultured human nucleus pulposus cells (NPCs) and vascular endothelial cells (VECs) and regulated the expression of SDF1/CXCR7 to investigate the effect of VEC angiogenesis by NPCs. The results revealed that angiogenesis was enhanced with increased SDF1 and that angiogenesis was weakened with the inhibition of CXCR7. We found that PI3K/AKT was involved in the downstream pathway in the coculture. VEC angiogenesis induction by NPCs was enhanced with an increase in pAKT or a decrease in PTEN. We conclude that the SDF1/CXCR7 signaling axis plays a role in the angiogenesis of degenerated IVD through the PI3K/AKT pathway.


Assuntos
Quimiocina CXCL12/metabolismo , Degeneração do Disco Intervertebral/patologia , Neovascularização Patológica/patologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptores CXCR/metabolismo , Idoso , Células Cultivadas , Quimiocina CXCL12/genética , Técnicas de Cocultura , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Feminino , Humanos , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/metabolismo , Masculino , Pessoa de Meia-Idade , Neovascularização Patológica/genética , Neovascularização Patológica/metabolismo , Núcleo Pulposo/metabolismo , Núcleo Pulposo/patologia , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/genética , Receptores CXCR/genética , Transdução de Sinais
9.
Biomed Pharmacother ; 113: 108652, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30856535

RESUMO

Emerging evidence suggests that microRNAs (miRNAs, miRs) play important roles in the development of intervertebral disc degeneration (IVDD). Nonetheless, the expression level and biological function of miR-499a-5p in IVDD are still unclear. In this study, we found that miR-499a-5p was significantly downregulated in degenerative tissues of the human nucleus pulposus (NP) compared with healthy tissues. Knockdown of miR-499a-5p promoted NP cell (NPC) apoptosis, stimulated caspase activation, enhanced MMP3 and MMP13 expression, and downregulated aggrecan and type II collagen. Furthermore, TNF-α-treated NPCs showed increased apoptosis and induced an imbalance between anabolism and catabolism of the extracellular matrix (ECM); these changes were attenuated by miR-499a-5p overexpression. Research into possible mechanisms revealed that miR-499a-5p suppressed the expression of SOX4 both at mRNA and protein levels and directly bound to the 3' untranslated region of SOX4 mRNA. Ectopic expression of SOX4 attenuated the negative effect of miR-499a-5p on NPC apoptosis and the positive effect on ECM synthesis. Taken together, these results indicate that miR-499a-5p may attenuate TNF-α-induced NPC apoptosis and an imbalance between anabolism and catabolism of the ECM by downregulating SOX4.


Assuntos
Apoptose/genética , Matriz Extracelular/patologia , Degeneração do Disco Intervertebral/patologia , MicroRNAs/genética , Núcleo Pulposo/patologia , Fatores de Transcrição SOXC/metabolismo , Regiões 3' não Traduzidas , Células Cultivadas , Regulação para Baixo , Matriz Extracelular/metabolismo , Feminino , Humanos , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/metabolismo , Masculino , Pessoa de Meia-Idade , Núcleo Pulposo/metabolismo
10.
Life Sci ; 221: 274-283, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30797016

RESUMO

AIMS: Lower back pain is often associated with intervertebral disc degeneration (IDD), which results from a decrease in nucleus pulposus (NP) cells and an imbalance between the degradation and synthesis of extracellular matrix (ECM) components. Multiple microRNAs play crucial roles in the modulation of NP cell apoptosis and matrix degradation. miR-145 is an important microRNA related to degenerative diseases such as osteoarthritis. Here, the effect of miR-145 in IDD was elucidated. The aim of this study was to explore the role and mechanism of miR-145 in the apoptosis of NP cells and in matrix metabolism in NP cells. MATERIALS AND METHODS: Real-time PCR, western blotting and flow cytometry analysis were used to observe the effect of miR-145 on NP cell apoptosis in the absence or presence of oxidative stress. Cell transfection, loss-of-function experiments using an ADAM17 inhibitor or lentiviral shADAM17, immunofluorescence, real-time PCR and western blotting were performed to demonstrate the role and mechanism of miR-145 in NP cell matrix metabolism. KEY FINDINGS: miR-145 attenuated NP cell apoptosis in the absence and presence of oxidative stress. Moreover, miR-145 overexpression increased and miR-145 suppression decreased matrix synthesis. ADAM17, which is expressed in degenerative discs, is the target of miR-145. ADAM17 gene suppression with lentiviral shRNA or an inhibitor enhanced matrix synthesis in NP cells. In addition, siADAM17 reversed the matrix degradation induced by miR-145 inhibition. SIGNIFICANCE: miR-145 suppresses apoptosis and promotes ECM synthesis in NP cells. miR-145 is thus a potential therapeutic microRNA for IDD.


Assuntos
MicroRNAs/genética , MicroRNAs/metabolismo , Núcleo Pulposo/metabolismo , Proteína ADAM17/genética , Proteína ADAM17/metabolismo , Animais , Apoptose/genética , Células Cultivadas , Matriz Extracelular/genética , Matriz Extracelular/metabolismo , Humanos , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/metabolismo , Ratos , Ratos Sprague-Dawley
11.
DNA Cell Biol ; 38(4): 358-366, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30758228

RESUMO

Intervertebral disk degeneration (IDD) is a common disease that is caused by degeneration of the nucleus pulposus (NP). One goal in the treatment of IDD is delaying or reversing the degeneration of NP via the transformation of exogenous genes. This study first investigated the role of BMP9 in the extracellular matrix (ECM) of nucleus pulposus cells (NPCs) and its mechanism. We found that BMP9 promotes the expression of ECM in NPCs, and the key molecules of Notch signaling, namely, NICD-1, hes and hey, and it was significantly altered in BMP9-transfected NPCs, which suggests that BMP9 may regulate the ECM via the Notch signaling pathway. We verified the expression of Notch ligands and receptors in NPCs infected with Ad-BMP9 and demonstrated a significant decrease in DLL1 and Notch1; then, NPCs were transfected with Ad-dnNotch1, Ad-Jagged1, and Ad-DLL1, and different multiple groups were established to further identify the ligands or receptors that affected ECM expression. The results demonstrated that Ad-dnNotch1, Jagged1 and DLL1 inhibited ECM expression, and dnNotch1 promoted expression. Therefore, we demonstrated that BMP9 promoted the expression of ECM in NPCs via inhibition of Notch1 and DLL1. This study provides a possible method for IDD treatment.


Assuntos
Matriz Extracelular/metabolismo , Fatores de Diferenciação de Crescimento/metabolismo , Núcleo Pulposo/patologia , Receptor Notch1/metabolismo , Transdução de Sinais , Proliferação de Células , Feminino , Regulação da Expressão Gênica , Fatores de Diferenciação de Crescimento/genética , Humanos , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/metabolismo , Degeneração do Disco Intervertebral/patologia , Masculino , Pessoa de Meia-Idade
12.
Int J Mol Sci ; 20(3)2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30717203

RESUMO

Ozone therapy has been widely used in everyday clinical practice over the last few years, leading to significant clinical results in the treatment of herniated discs and pain management. Nevertheless, further studies have demonstrated its potential efficacy and safety under other clinical and experimental conditions. However, some of these studies showed controversial results regarding the safety and efficacy of ozone therapy, thus mining its potential use in an everyday clinical practice. To this regard, it should be considered that extensive literature review reported the use of ozone in a significant different dose range and with different delivery systems. The aim of the present review is to describe the various pharmacological effects of ozone in different organs and clinical conditions and to provide possible biochemical and molecular insights for ozone biological properties, thus providing a possible explanation for various controversial clinical outcomes described in the scientific literature.


Assuntos
Doenças Cardiovasculares/terapia , Degeneração do Disco Intervertebral/terapia , Deslocamento do Disco Intervertebral/terapia , Ozônio/administração & dosagem , Dor/prevenção & controle , Substâncias Protetoras/administração & dosagem , Dermatopatias/terapia , Doença Aguda , Doenças Cardiovasculares/genética , Doenças Cardiovasculares/imunologia , Doenças Cardiovasculares/patologia , Quimiotaxia/efeitos dos fármacos , Quimiotaxia/imunologia , Doença Crônica , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Humanos , Imunidade Inata/efeitos dos fármacos , Disco Intervertebral/efeitos dos fármacos , Disco Intervertebral/imunologia , Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/imunologia , Degeneração do Disco Intervertebral/patologia , Deslocamento do Disco Intervertebral/genética , Deslocamento do Disco Intervertebral/imunologia , Deslocamento do Disco Intervertebral/patologia , Estresse Oxidativo , Ozônio/efeitos adversos , Dor/genética , Dor/imunologia , Dor/patologia , Manejo da Dor/métodos , Substâncias Protetoras/efeitos adversos , Dermatopatias/genética , Dermatopatias/imunologia , Dermatopatias/patologia
13.
In Vivo ; 33(2): 413-417, 2019 Mar-Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30804119

RESUMO

BACKGROUND/AIM: The present study aimed to investigate the role of an aggrecan (ACAN) gene variant and proteoglycan levels in the risk of lumbar degenerative disc disease (LDDD). MATERIALS AND METHODS: A total of 108 patients with LDDD and 103 healthy controls were enrolled. Molecular assessment of the ACAN gene (c.6423T>C) variant was determined by real time-polymerase chain reaction. Proteoglycan levels in serum were measured with enzyme-linked immunosorbent assay. RESULTS: The frequency of all alleles and genotypes in all study groups were distributed according to the Hardy-Weinberg equilibrium. In addition, no association between the ACAN gene (c.6423T>C) variant and presence of risk factors for LDDD was detected. However, proteoglycan levels were significantly lower in patients with LDDD compared to the control group (p<0.00001). CONCLUSION: Our findings suggest that proteoglycan has emerged as a potential novel biomarker which might be used for prediction of LDDD risk.


Assuntos
Agrecanas/genética , Predisposição Genética para Doença , Degeneração do Disco Intervertebral/genética , Deslocamento do Disco Intervertebral/genética , Alelos , Grupo com Ancestrais do Continente Asiático/genética , Feminino , Estudos de Associação Genética , Genótipo , Humanos , Degeneração do Disco Intervertebral/sangue , Degeneração do Disco Intervertebral/fisiopatologia , Deslocamento do Disco Intervertebral/sangue , Deslocamento do Disco Intervertebral/fisiopatologia , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único/genética , Proteoglicanas/sangue , Proteoglicanas/genética
14.
Int J Mol Sci ; 20(3)2019 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-30717434

RESUMO

The role of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) has been highlighted in mechanisms underlying inflammatory and neuropathic pain processes. The present study was designed to investigate whether NF-κB signaling is associated with pain-related neuropeptide expression in patients with chronic back pain related to degenerative disc disease (DDD). Intervertebral disc (IVD) tissues were collected from forty DDD patients undergoing disc replacement or fusion surgery, and from eighteen postmortem (PM) control subjects. RELA, NFKB1, CGRP, TAC1, TRPV1, and MMP-3 gene expression were analyzed by RT-qPCR, while NF-κB subunit RelA and NF-κB1⁻DNA binding in nuclear extracts and calcitonin gene related peptide (CGRP), substance P (SP), and transient receptor potential, subfamily V, member 1 (TRPV1) protein levels in cytosolic extracts of tissues were assessed by enzyme-linked immunosorbent assay (ELISA). An upregulated NF-κB1⁻DNA binding, and higher CGRP and TRPV1 protein levels were observed in DDD patients compared to PM controls. In DDD patients, NF-κB1⁻DNA binding was positively correlated with nuclear RelA levels. Moreover, NF-κB1⁻DNA binding was positively associated with TRPV1 and MMP-3 gene and SP and TRPV1 protein expression in DDD patients. Our results indicate that the expression of SP and TRPV1 in IVD tissues was associated with NF-κB activation. Moreover, NF-κB may be involved in the generation or maintenance of peripheral pain mechanisms by the regulation of pain-related neuropeptide expression in DDD patients.


Assuntos
Degeneração do Disco Intervertebral/metabolismo , NF-kappa B/metabolismo , Dor/metabolismo , Transdução de Sinais , Substância P/genética , Canais de Cátion TRPV/genética , Adulto , Feminino , Regulação da Expressão Gênica , Humanos , Disco Intervertebral/metabolismo , Disco Intervertebral/fisiopatologia , Degeneração do Disco Intervertebral/complicações , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/fisiopatologia , Masculino , Pessoa de Meia-Idade , NF-kappa B/fisiologia , Subunidade p50 de NF-kappa B/metabolismo , Subunidade p50 de NF-kappa B/fisiologia , Dor/etiologia , Dor/genética , Fator de Transcrição RelA/metabolismo , Fator de Transcrição RelA/fisiologia
15.
Mol Med Rep ; 19(3): 2440-2448, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30664184

RESUMO

Increasing cell apoptosis is one of the major causes of intervertebral disc degeneration (IDD). ß-ecdysterone has been demonstrated to protect PC12 cells against neurotoxicity. A previous study revealed that ß­ecdysterone may be involved in the regulation of autophagy in osteoblasts. Therefore, we hypothesized that ß­ecdysterone may possess therapeutic effects on IDD via autophagy stimulation. The effect of ß­ecdysterone on IDD was explored by in vitro experiments. The results demonstrated that ß­ecdysterone attenuated the apoptosis induced by tert­butyl hydroperoxide via promoting autophagy in nucleus pulposus cells. Beclin­1, an indispensable protein for the stimulation of autophagy, is upregulated and stabilized by ß­ecdysterone in a dose­ and time­dependent manner in nucleus pulposus cells. Inhibition of autophagy with 3­methyladenine partially abrogated the protective function of ß­ecdysterone against apoptosis of nucleus pulposus cells, indicating that autophagy participated in the protective effect of ß­ecdysterone on IDD. Additionally, ß­ecdysterone promoted the expression of anabolic genes while inhibiting the expression of catabolic genes in nucleus pulposus cells. Collectively, the present study demonstrated that ß­ecdysterone may protect nucleus pulposus cells against apoptosis by autophagy stimulation and ameliorate disc degeneration, which indicates that ß­ecdysterone may be a potential therapeutic agent for IDD.


Assuntos
Autofagia/efeitos dos fármacos , Proteína Receptora de AMP Cíclico/administração & dosagem , Ecdisterona/administração & dosagem , Degeneração do Disco Intervertebral/tratamento farmacológico , Núcleo Pulposo/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Proteína Beclina-1/genética , Ecdisterona/genética , Humanos , Disco Intervertebral/efeitos dos fármacos , Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/fisiopatologia , Núcleo Pulposo/metabolismo , Núcleo Pulposo/patologia , Osteoblastos/efeitos dos fármacos , Substâncias Protetoras/administração & dosagem , Ratos , Congêneres da Testosterona/biossíntese , Congêneres da Testosterona/genética
16.
Mol Med Rep ; 19(3): 2377-2385, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30664218

RESUMO

Leukemia inhibitory factor (LIF) is a multifunctional cytokine. The present study aimed to determine the expression and effects of LIF on nucleus pulposus generation. Degenerated nucleus pulposus samples were obtained from animal models and patients with lumbar intervertebral disc herniation. Degradation scores of intervertebral discs were evaluated via magnetic resonance imaging (MRI) and histology, and the protein expression levels of LIF were detected. Furthermore, cultured primary human degenerated nucleus pulposus cells (DNPCs) were stimulated with various concentrations of recombinant human LIF protein (rhLIF), and aggrecan and collagen type II α1 (COL2α1) protein expression levels were detected by western blotting. In addition, aggrecan expression was determined by toluidine blue staining. The effects of rhLIF on proliferation and apoptosis of DNPCs were evaluated by Cell Counting Kit­8 and flow cytometry, respectively. The results revealed that the degradation scores of intervertebral discs were significantly associated with modeling time, as determined by MRI and histology. In addition, the protein expression levels of LIF were initially increased in patients with lumbar disc herniation and in rabbit models, particularly in the 2­week modeling group; however, its expression decreased with the progression of disc degeneration. Notably, LIF expression in each modeling group was higher than that in the control and 0 week modeling group. The in vitro study revealed that the protein expression levels of aggrecan and COL2α1 were significantly increased in response to rhLIF, in a dose­dependent manner, and statistical differences were identified between the treatment groups and control group. The results of toluidine blue staining were consistent with this finding. Although rhLIF had no effect on proliferation, it inhibited apoptosis of DNPCs in a concentration­dependent manner. In conclusion, LIF was upregulated during the process of intervertebral disc degeneration, and may promote the expression of extracellular matrix components. It may also be hypothesized that LIF acts as a potential protective factor by inhibiting apoptosis of DNPCs without affecting cell proliferation.


Assuntos
Degeneração do Disco Intervertebral/tratamento farmacológico , Fator Inibidor de Leucemia/genética , Núcleo Pulposo/efeitos dos fármacos , Proteínas Recombinantes/administração & dosagem , Agrecanas/genética , Animais , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Colágeno Tipo II/genética , Feminino , Citometria de Fluxo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Disco Intervertebral/diagnóstico por imagem , Disco Intervertebral/metabolismo , Disco Intervertebral/fisiopatologia , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/fisiopatologia , Deslocamento do Disco Intervertebral/tratamento farmacológico , Deslocamento do Disco Intervertebral/genética , Deslocamento do Disco Intervertebral/fisiopatologia , Fator Inibidor de Leucemia/administração & dosagem , Imagem por Ressonância Magnética , Masculino , Núcleo Pulposo/diagnóstico por imagem , Núcleo Pulposo/crescimento & desenvolvimento , Núcleo Pulposo/patologia , Coelhos , Proteínas Recombinantes/genética
17.
Mol Genet Genomic Med ; 7(2): e00524, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30548218

RESUMO

BACKGROUND: Lumbar disk disease (LDD) is a common musculoskeletal disorder. Several predisposing genetic and environmental risk factors have been established for symptomatic LDD. METHODS: We conducted a case-control association study to investigate the role of the COL11A2 gene in LDD. Genotyping of 384 Chinese Han LDD patients and 384 Chinese Han controls was made for six single-nucleotide polymorphisms (SNPs) from COL11A2 by Agena Massarray. We evaluated these SNPs association with LDD using the chi-square test and genetic model analysis. RESULTS: The strongest associations with LDD were observed for polymorphisms in rs2071025. Carriers of "A" allele had an increased risk of LDD (OR = 1.47, 95% CI = 1.20-1.80, p = 0.0002) as compared with the "G" allele in allele model. We found that rs2071025 were associated with LDD in female and male from the stratification analyses (p < 0.05). Genetic models showed that rs986522(C) significantly increased the risk of LDD in female; however, in males, we did not find significant associations between the rs986522 and LDD risk. CONCLUSION: This study showed a genetic association with COL11A2 polymorphism in individuals with LDD. These data may provide novel insights into the pathogenesis of LDD, although further studies with larger numbers of participants worldwide are needed for validation of our conclusions.


Assuntos
Colágeno Tipo XI/genética , Degeneração do Disco Intervertebral/genética , Polimorfismo de Nucleotídeo Único , China , Feminino , Humanos , Vértebras Lombares/patologia , Masculino , Pessoa de Meia-Idade
18.
Cell Mol Biol (Noisy-le-grand) ; 64(10): 61-65, 2018 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-30084796

RESUMO

Intervertebral disc degeneration (IVDD) is a common degenerative spinal condition. Recent studies have shown that the incidence of disc herniation and disc degeneration may be explained by genetic factors.  In this study, we investigated the link between various polymorphic variants of the vitamin D receptor (VDR), matrix metalloproteinase 2 (MMP2), and insulin like growth factor 1 receptor (IGF1R) genes and IVDD in patients with IVDD, in Turkey. We examined and genotyped 199 patients with IVDD and 197 healthy individuals. Genomic DNA was isolated from the peripheral blood leukocytes of all participants, and analyzed using real-time PCR. Via melting curve analysis, VDR, MMP2, and IGF1R polymorphism variant distributions were determined. The patients with IVDD showed higher frequencies of the VDR ApaI A allele genotype as compared to the control group; however, there were no significant differences in the frequencies or allelic distributions of the IGF1R and MMP2 genotypes between the IVDD patients and the control group. The incidence of IVDD in these Turkish patients is correlated with the VDR ApaI gene polymorphism, but not with the IGF1R and MMP2 polymorphisms.


Assuntos
Degeneração do Disco Intervertebral/genética , Metaloproteinase 2 da Matriz/genética , Polimorfismo Genético , Receptores de Calcitriol/genética , Receptores de Somatomedina/genética , Adulto , Feminino , Genótipo , Humanos , Degeneração do Disco Intervertebral/epidemiologia , Masculino , Pessoa de Meia-Idade , Turquia/epidemiologia
19.
Med Sci Monit ; 24: 5598-5609, 2018 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-30099472

RESUMO

BACKGROUND The present study aimed to evaluate whether the fat mass and obesity-associated (FTO) gene polymorphisms are associated with risk of intervertebral disc degeneration (IDD) in a largest Chinese Han population. MATERIAL AND METHODS There were 502 IDD patients and 497 healthy controls enrolled in this study. Nineteen single nucleotide polymorphisms (SNPs) in the FTO gene were tested using the Sequenom MassARRAY platform. The Hardy-Weinberg equilibrium test, followed by allelic, genotypic, haplotypic association, and SNP interaction analyses were used for SNP evaluation. The Genotype-Tissue Expression (GTEx) database was used to evaluate expression quantitative trait loci (eQTL) value of polymorphism. Spearman rank correlation and logistic regression analyses were used for assessing the internal relation between genotypic changes and the risk of IDD. RESULTS Seventeen SNPs survived the Hardy-Weinberg equilibrium test. Allelic analysis showed that allele T of SNP rs1121980 was a risk allele. Haplotypic and SNP interaction analyses suggested that 2 haplotypes and 5 SNP combinations were associated with the predisposition of IDD respectively. GTEx database revealed that the SNP rs1121980 might interfere with the expression of the FTO gene in the muscle-skeletal system. Through clinical statistics analysis, the different genotypes of rs1121980 can present different disease severity of IDD. CONCLUSIONS Our study suggests that rs1121980 can become a biomarker for the screening and prognosis of IDD. The 2 haplotype blocks and 5 SNP-SNP combinations that we discovered might be indicative of the onset of IDD. Therefore, our study might serve as evidence for future IDD molecular diagnosis.


Assuntos
Dioxigenase FTO Dependente de alfa-Cetoglutarato/genética , Degeneração do Disco Intervertebral/genética , Adulto , Dioxigenase FTO Dependente de alfa-Cetoglutarato/metabolismo , Grupo com Ancestrais do Continente Asiático/genética , Estudos de Casos e Controles , China , Grupos Étnicos/genética , Feminino , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Haplótipos , Humanos , Degeneração do Disco Intervertebral/metabolismo , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único/genética , Fatores de Risco
20.
Cell Prolif ; 51(5): e12483, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30039593

RESUMO

Intervertebral disc degeneration (IDD) is the major cause of low back pain which incurs a significant public-health and economic burden. The aetiology of IDD is complex, with developmental, genetic, biomechanical and biochemical factors contributing to the disease development. Deregulated phenotypes of nucleus pulposus cells, including aberrant differentiation, apoptosis, proliferation and extracellular matrix deposition, are involved in the initiation and progression of IDD. Non-coding RNAs, including long non-coding RNAs (lncRNAs), have recently been identified as important regulators of gene expression. Research into their roles in IDD has been very active over the past 5 years. Our review summarizes current research regarding the roles of deregulated lncRNAs (eg, RP11-296A18.3, TUG1, HCG18) in modulating nucleus pulposus cell functions in IDD. These exciting findings suggest that specific modulation of lncRNAs or their downstream signalling pathways might be an attractive approach for developing novel therapeutics for IDD.


Assuntos
Degeneração do Disco Intervertebral/genética , Núcleo Pulposo/metabolismo , RNA Longo não Codificante/genética , Apoptose/genética , Diferenciação Celular/genética , Expressão Gênica/genética , Humanos , Transdução de Sinais/genética
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