scholarly journals High Glucose-Induced Oxidative Stress Mediates Apoptosis and Extracellular Matrix Metabolic Imbalances Possibly via p38 MAPK Activation in Rat Nucleus Pulposus Cells

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Xiaofei Cheng ◽  
Bin Ni ◽  
Feng Zhang ◽  
Ying Hu ◽  
Jie Zhao
Keyword(s):  
Oxidative Stress ◽  
Extracellular Matrix ◽  
Nucleus Pulposus ◽  
P38 Mapk ◽  
High Glucose ◽  
Type Ii Collagen ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Type Ii ◽  
Nucleus Pulposus Cells ◽  
Mapk Activation

Objectives. To investigate whether high glucose-induced oxidative stress is implicated in apoptosis of rat nucleus pulposus cells (NPCs) and abnormal expression of critical genes involved in the metabolic balance of extracellular matrix (ECM).Methods. NPCs were cultured with various concentrations of glucose to detect cell viability and apoptosis. Cells cultured with high glucose (25 mM) were untreated or pretreated with N-acetylcysteine or a p38 MAPK inhibitor SB 202190. Reactive oxygen species (ROS) production was evaluated. Activation of p38 MAPK was measured by Western blot. The expression of ECM metabolism-related genes, including type II collagen, aggrecan, SRY-related high-mobility-group box 9 (Sox-9), matrix metalloproteinase 3 (MMP-3), and tissue inhibitor of metalloproteinase 1 (TIMP-1), was analyzed by semiquantitative RT-PCR.Results. High glucose reduced viability of NPCs and induced apoptosis. High glucose resulted in increased ROS generation and p38 MAPK activation. In addition, it negatively regulated the expression of type II collagen, aggrecan, Sox-9, and TIMP-1 and positively regulated MMP-3 expression. These results were changed by pretreatment with N-acetylcysteine or SB 202190.Conclusions. High glucose might promote apoptosis of NPCs, trigger ECM catabolic pathways, and inhibit its anabolic activities, possibly through a p38 MAPK-dependent oxidative stress mechanism.

scholarly journals Research on Roles of Mongolian Medical Warm Acupuncture in Inhibiting p38 MAPK Activation and Apoptosis of Nucleus Pulposus Cells

2018 ◽  
Vol 2018 ◽  
pp. 1-8
Author(s):  
Sha Li ◽  
Lidao Bao ◽  
Lengge Si ◽  
Xiaohui Wang ◽  
Agula Bo
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Extracellular Matrix ◽  
Matrix Metalloproteinase ◽  
Nucleus Pulposus ◽  
P38 Mapk ◽  
Mapk Pathway ◽  
Active Oxygen ◽  
Nucleus Pulposus Cells ◽  
P38 Mapk Pathway

Background. Mongolian medical warm acupuncture has a desirable therapeutic effect on sciatica. Apoptosis of the nucleus pulposus cells is considered to play an important role in sciatica. Evidence has demonstrated that oxidative stress and its induced activation of the signaling pathways play important roles in sciatica. However, further research is expected to reveal whether Mongolian medical warm acupuncture can inhibit the apoptosis of nucleus pulposus cells and oxidative stress. Objective. To study the effect of the p38 MAPK pathway activated by the generated ROS on apoptosis and the expression of the genes related to the balance of the extracellular matrix metabolism during treatment of sciatica with Mongolian medical warm acupuncture. Method. The volume of the active oxygen generated in the nucleus pulposus cells was detected following intervention of Mongolian medical warm acupuncture. The p38 MAPK phosphorylation level was detected with Western blot. The genes are related to the metabolism of the nucleus pulposus extracellular matrix. Result. Mongolian medical warm acupuncture reduced the active oxygen within the nucleus pulposus cells and inhibited the activation of the p38 MAPK pathway (P=0.013). Meanwhile, it upregulated the gene expression of Type II collagen, aggrecan, Sox-9, and tissue matrix metalloproteinase reagent 1 (P-0.015; P=0.025; P=0.031; P=0.045) and downregulated the gene expression of matrix metalloproteinase 3 (P=0.015). Conclusion. Mongolian medical warm acupuncture may inhibit apoptosis of nucleus pulposus cells and activation of the extracellular matrix decomposition metabolism pathway and promote its anabolism. This process may rely on the oxidative stress matrix of the p38 MAPK pathway.

Lupeol against high-glucose-induced apoptosis via enhancing the anti-oxidative stress in rabbit nucleus pulposus cells

2018 ◽  
Vol 27 (10) ◽  
pp. 2609-2620 ◽  
Author(s):  
Ming-Bo Guo ◽  
De-Chun Wang ◽  
Hai-Fei Liu ◽  
Long-Wei Chen ◽  
Jian-Wei Wei ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Nucleus Pulposus ◽  
High Glucose ◽  
Nucleus Pulposus Cells ◽  
Induced Apoptosis

scholarly journals MicroRNA-222 regulates fluid shear stress-induced human nucleus pulposus cells degeneration through affecting c-Fos expression

2019 ◽  
Author(s):  
Haixiong Miao ◽  
Yicun Yao ◽  
Baoqing Ye ◽  
Libing Dai ◽  
Weiguo Liang
Keyword(s):  
Shear Stress ◽  
Molecular Mechanism ◽  
Nucleus Pulposus ◽  
Fluid Shear Stress ◽  
Type Ii Collagen ◽  
Cell Counting ◽  
Type Ii ◽  
Fluid Shear ◽  
Nucleus Pulposus Cells

AbstractIntervertebral disc degeneration (IVDD) is a chronic disease that correlates with the deterioration of the nucleus pulposus (NP) cells. However, the molecular mechanism of IVDD remains unclear. In this study, we investigated the function of microRNA-222 in IVDD and the potential molecular mechanism. NP cells treated with fluid shear stress (FSS) were used to simulate a model of IVDD in vitro. MicroRNA-222 was significantly downregulated in NP cells stimulated with FSS compared with that in unstimulated NP cells. Human NP cells were also treated with FSS to induce their degeneration. The mRNA and protein levels of C-FOS, MEK, phosphorylated MEK5 (pMEK5), ERK5, and pERK5 were evaluated with RT-PCR and western blotting, respectively. Enzyme-linked immunosorbent assays were used to investigate type II collagen and Aggrecan expression. NP cell proliferation was determined with the Cell Counting Kit-8. MicroRNA-222 was significantly downregulated in NP cells treated with FSS. The production of c-Fos and MEK5 were markedly reduced or increased in NP cells transfected with the has-microRNA-222 mimic or inhibitor, respectively, whether or not they were stimulated with FSS. The overexpression or inhibition of microRNA-222 markedly accelerated or suppressed the apoptosis of FSS-stimulated NP cells, respectively. In the NP cells, the overexpression or inhibition of microRNA-222 massively inhibited or strengthened Aggrecan and type II collagen expression. Together, our data indicated that c-Fos was a target of microRNA-222, and was negatively regulated by microRNA-222 in NP cells. Our findings also suggested that microRNA-222 is a possible therapeutic target for IVDD because it regulates c-Fos.

Inhibition of proteoglycan and type II collagen synthesis of disc nucleus cells by nicotine

2003 ◽  
Vol 99 (3) ◽  
pp. 291-297 ◽  
Author(s):  
Keun Su Kim ◽  
S. Tim Yoon ◽  
Jin Soo Park ◽  
Jun Li ◽  
Moon Soo Park ◽  
...  
Keyword(s):  
Phase I ◽  
Nucleus Pulposus ◽  
Phase Ii ◽  
Phase I Study ◽  
Phase Ii Study ◽  
Type Ii Collagen ◽  
Type Ii ◽  
Nucleus Pulposus Cells ◽  
Content Type ◽  
Fine Print

Object. Systemic nicotine has been hypothesized to cause degeneration of the intervertebral disc which in turn decreases vascular supply to the disc through a cholinergic receptor—mediated process. Another possible mechanism may be through direct regulatory effects on disc cells. In this study, the authors tested the hypothesis that nicotine adversely affects nucleus pulposus cells by directly inhibiting proteoglycan synthesis and gene expression of type II collagen (Phase I study). They also assessed the hypothesis that nicotine inhibits the bone morphogenetic protein (BMP)—2-induced upregulation of extracellular matrix (Phase II study). Methods. Cells were isolated from nucleus pulposus obtained in rat lumbar discs and cultured on a monolayer. Media were treated with nicotine and/or recombinant human (rh)BMP-2 for 7 days. Sulfated glycosaminoglycan (SO4-GAG) in media was quantified using 1,9-dimethylmethylene blue (DMMB) assay. Gene assay of types I and II collagen, Sox9, and glyceraldehyde-3-phosphate dehydrogenase were quantified using reverse transcriptase—polymerase chain reaction (RT-PCR) and real time PCR. In the Phase I study, nicotine-treated (100 µg/ml) and nontreated cells were compared. The s-GAG production and messenger RNA (mRNA) of type II collagen and Sox9 decreased significantly in the nicotine-treated group. In the Phase II study, five groups were compared: 1) nontreatment; 2) rhBMP-2 only (100 ng/ml); and 3–5) with rhBMP-2 (100 ng/ml) and increasing doses of nicotine (1 [third group], 10, [fourth group], 100 [fifth group] µg/ml). The SO4-GAG production and mRNA of type II collagen and Sox9 decreased significantly in the groups treated with rhBMP-2 combined with 10 and 100 µg/ml of nicotine compared with the group treated with rhBMP-2. Conclusions. The results of this study raise the possibility that nicotine may contribute to the process of disc degeneration by a direct effect on the nucleus pulposus cells, possibly by antagonizing the effect of BMP-2.

Effect of Matrix Metalloproteinase 3 Gene Silencing and SRY-Related High Mobility Group-Box Gene 9 Gene Overexpression on Human Degeneration Nucleus Pulposus Cells In Vitro

2019 ◽  
Vol 9 (5) ◽  
pp. 707-713
Author(s):  
Yong-Ming Xi ◽  
Xiang-Zhen Jia ◽  
Tao Yu ◽  
Hua-Wei Wei ◽  
Hua-Cong Wang ◽  
...  
Keyword(s):  
Intervertebral Disc ◽  
Gene Silencing ◽  
Nucleus Pulposus ◽  
Type Ii Collagen ◽  
Gene Overexpression ◽  
Control Group ◽  
Type Ii ◽  
Nucleus Pulposus Cells ◽  
Vector Group ◽  
Sox9 Gene

This study aims to detect the biological effects of matrix metalloproteinase 3 (MMP3) and SRY-related high mobility group-box gene 9 (SOX9) gene regulation in the human intervertebral disc degeneration of nucleus pulposus cells mediated by lentiviral vectors in vitro. The culture and development of human degenerated intervertebral disc nucleus pulposus cells was performed. The cell morphology and structure were observed, as identified by hematoxylin and eosin (H&E) staining. The experiment was divided into five groups: blank control group, SOX9 gene overexpression group, MMP3 gene silencing group, SOX9 gene overexpression+MMP3 gene silencing group, and blank vector group. RT-PCR and western blot were performed to detect the influence of MMP3 gene silencing and SOX9 gene overexpression on degenerated human intervertebral disc nucleus pulposus cells, include the secretion of polysaccharide and expression of type II collagen, at the mRNA and protein level. The originally generated degenerated human intervertebral disc nucleus pulposus cells had the same cellular morphology. The MTT assay revealed that the blank group and blank vector group had no statistical significance in cell proliferation. The MMP3 gene silencing and SOX9 gene overexpression could promote the proliferation of degenerated human intervertebral disc nucleus pulposus cells, when compared with the blank control group and blank vector group, and this more obvious in the MMP3 gene silencing group and SOX9 gene overexpression group. RT-PCR and western blot revealed that MMP3 gene silencing and SOX9 gene overexpression can promote cells to secrete polysaccharide and type-II collagen (P < 0.05). MMP3 gene silencing and SOX9 gene overexpression can promote the proliferation of degenerated human intervertebral disc nucleus pulposus cells, and their ability to secrete polysaccharide and type-II collagen.

scholarly journals Human Bone Marrow Mesenchymal Stromal Cell-Derived Extracellular Vesicles Promote Proliferation of Degenerated Nucleus Pulposus Cells and the Synthesis of Extracellular Matrix Through the SOX4/Wnt/β-Catenin Axis

2021 ◽  
Vol 12 ◽  
Author(s):  
Haoyu Wang ◽  
Fei Li ◽  
Wenrui Ban ◽  
Jing Zhang ◽  
Guiqi Zhang
Keyword(s):  
Extracellular Matrix ◽  
Nucleus Pulposus ◽  
Extracellular Vesicles ◽  
Target Genes ◽  
Intervertebral Disk ◽  
Human Bone ◽  
Nucleus Pulposus Cells ◽  
Synthetic Genes ◽  
Downstream Target ◽  
Transcription Factor 4

Objective: Intervertebral disk degeneration (IDD) is a major cause of pain in the back, neck, and radiculus. Mesenchymal stem cells (MSCs)-derived extracellular vesicles (EVs) are therapeutic in musculoskeletal degenerative diseases such as IDD. This study explored the effect and functional mechanism of human bone MSCs (hBMSCs)-derived EVs in proliferation and apoptosis of degenerated nucleus pulposus cells (DNPCs) and extracellular matrix (ECM) synthesis.Methods: Extracellular vesicles were isolated from hBMSCs and identified. DNPCs were induced by TNF-α. EVs were incubated with DNPCs for 24h. Internalization of EVs by DNPCs, DNPCs proliferation, apoptosis, and expressions of ECM synthetic genes, degrading genes and miR-129-5p were assessed. Downstream target genes of miR-129-5p were predicted. Target relation between miR-129-5p and SRY-box transcription factor 4 (SOX4) was verified. DNPCs proliferation, apoptosis, and ECM synthesis were measured after treatment with EVs and miR-129-5p inhibitor or SOX4 overexpression. Expressions of SOX4 and Wnt/β-catenin pathway-related proteins were determined.Results: hBMSC-EVs promoted DNPCs proliferation, inhibited apoptosis, increased expressions of ECM synthetic genes, and reduced expressions of ECM degrading genes. hBMSC-EVs carried miR-129-5p into DNPCs. Silencing miR-129-5p in EVs partially inverted the effect of EVs on DNPCs proliferation and ECM synthesis. miR-129-5p targeted SOX4. SOX4 overexpression annulled the effect of EVs on DNPCs proliferation and ECM synthesis. Expressions of Wnt1 and β-catenin were decreased in EVs-treated DNPCs, while silencing miR-129-5p in EVs promoted expressions of Wnt1 and β-catenin.Conclusion: hBMSC-EVs promoted DNPCs proliferation and ECM synthesis by carrying miR-129-5p into DNPCs to target SOX4 and deactivating the Wnt/β-catenin axis.

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