scholarly journals Resveratrol enhances matrix biosynthesis of nucleus pulposus cells through activating autophagy via the PI3K/Akt pathway under oxidative damage

2018 ◽  
Vol 38 (4) ◽  
Author(s):  
Jinlou Gao ◽  
Qingyun Zhang ◽  
Lin Song
Keyword(s):  
Oxidative Damage ◽  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
Akt Pathway ◽  
Control Group ◽  
Protective Effects ◽  
Nucleus Pulposus Cells ◽  
Matrix Synthesis ◽  
Cellular Autophagy ◽  
Matrix Production

The decrease in nucleus pulposus (NP) matrix production is a classic feature during disc degeneration. Resveratrol (RSV) is reported to play protective effects under many pathological factors.The present study aims to study the effects of RSV on NP matrix homeostasis under oxidative damage and the potential mechanism. Rat NP cells were exposed to H2O2 solution to create an oxidative damage. RSV and the 3-methyladenine (3-MA) were added along with the culture medium to respectively investigate the role of RSV and cellular autophagy. NP matrix synthesis was evaluated by the expression of macromolecules (aggrecan and collagen II) and glycosaminoglycan (GAG) content. Activation of cellular autophagy was assessed by the expression of several molecular markers. Additionally, activity of the PI3K/Akt pathway was also evaluated to study its potential role. Compared with the control group (NP cells treated with H2O2), RSV significantly up-regulated expression of matrix macromolecules (aggrecan and collagen), promoted GAG production, and increased the expression of autophagy-related markers (Beclin-1 and LC-3). Further analysis showed that inhibition of autophagy by 3-MA partly attenuated NP matrix production. Additionally, RSV increased activity of the PI3K/Akt pathway compared with the control NP cells, but it was not affected by the addition of 3-MA. RSV plays a protective role in enhancing NP matrix synthesis under oxidative damage. Mechanistically, activation of the cellular autophagy via the PI3K/Akt pathway may participate in this process. RSV may be an effective drug to attenuate oxidative stress-induced disc degeneration.

scholarly journals Icariin Prevents H2O2-Induced Apoptosis via the PI3K/Akt Pathway in Rat Nucleus Pulposus Intervertebral Disc Cells

2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
Author(s):  
Xiangyu Deng ◽  
Sheng Chen ◽  
Dong Zheng ◽  
Zengwu Shao ◽  
Hang Liang ◽  
...  
Keyword(s):  
Intervertebral Disc ◽  
Nucleus Pulposus ◽  
Caspase 3 ◽  
Chinese Herb ◽  
Akt Pathway ◽  
Control Group ◽  
Protective Effects ◽  
Nucleus Pulposus Cells ◽  
Intracellular Ros ◽  
Induced Apoptosis

Icariin is a prenylated flavonol glycoside derived from the Chinese herb Epimedium sagittatum. This study investigated the mechanism by which icariin prevents H2O2-induced apoptosis in rat nucleus pulposus (NP) cells. NP cells were isolated from the rat intervertebral disc and they were divided into five groups after 3 passages: (A) blank control; (B) 200 μM H2O2; (C) 200 μM H2O2 + 20 μM icariin; (D) 20 μM icariin + 200 μM H2O2 + 25 μM LY294002; (E) 200 μM H2O2 + 25 μM LY294002. LY294002 is a selective inhibitor of the phosphoinositide 3-kinase (PI3K)/Akt signaling pathway. NP cell viability, apoptosis rate, intracellular reactive oxygen species levels, and the expression of AKT, p-AKT, p53, Bcl-2, Bax, caspase-3 were estimated. The results show that, compared with the control group, H2O2 significantly increased NP cell apoptosis and the level of intracellular ROS. Icariin pretreatment significantly decreased H2O2-induced apoptosis and intracellular ROS and upregulated p-Akt and BCL-2 and downregulated caspase-3 and Bax. LY294002 abolished the protective effects of icariin. Our results show that icariin can attenuate H2O2-induced apoptosis in rat nucleus pulposus cells and PI3K/AKT pathway is at least partly included in this protection effect.

scholarly journals Melatonin Suppresses Apoptosis of Nucleus Pulposus Cells through Inhibiting Autophagy via the PI3K/Akt Pathway in a High-Glucose Culture

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Jian Li ◽  
Chengqiang Wang ◽  
Lixin Xue ◽  
Fan Zhang ◽  
Jianqiang Liu
Keyword(s):  
Protein Expression ◽  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
High Glucose ◽  
Cell Apoptosis ◽  
Caspase 3 ◽  
Akt Pathway ◽  
Protective Effects ◽  
Nucleus Pulposus Cells

Diabetes mellitus- (DM-) associated hyperglycemia promotes apoptosis of disc nucleus pulposus (NP) cells, which is a contributor to intervertebral disc degeneration (IDD). Melatonin is able to protect against cell apoptosis. However, its effects on apoptosis of NP cell in a high-glucose culture remain unclear. The purpose of the present study was to investigate the effects and molecular mechanism of melatonin on NP cell apoptosis in a high-glucose culture. NP cells were cultured in the baseline medium supplemented with a high-glucose concentration (0.2 M) for 3 days. The control cells were only cultured in the baseline medium. Additionally, the pharmaceutical inhibitor LY294002 was added along with the culture medium to investigate the possible role of the PI3K/Akt pathway. Apoptosis, autophagy, and activity of the PI3K/Akt pathway of NP cells among these groups were evaluated. Compared with the control NP cells, high glucose significantly increased cell apoptosis ratio and caspase-3/caspase-9 activity and decreased mRNA expression of Bcl-2, whereas it increased mRNA or protein expression of Bax, caspase-3, cleaved caspase-3, cleaved PARP, and autophagy-related molecules (Atg3, Atg5, Beclin-1, and LC3-II) and decreased protein expression of p-Akt compared with the control cells. Additionally, melatonin partly inhibited the effects of high glucose on those parameters of cell apoptosis, autophagy, and activation of PI3K/Akt. In conclusion, melatonin attenuates apoptosis of NP cells through inhibiting the excessive autophagy via the PI3K/Akt pathway in a high-glucose culture. This study provides new theoretical basis of the protective effects of melatonin against disc degeneration in a DM patient.

scholarly journals Dynamic Compression Promotes the Matrix Synthesis of Nucleus Pulposus Cells Through Up-Regulating N-CDH Expression in a Perfusion Bioreactor Culture

2018 ◽  
Vol 46 (2) ◽  
pp. 482-491 ◽  
Author(s):  
Yichun Xu ◽  
Hui Yao ◽  
Pei Li ◽  
Wenbin Xu ◽  
Junbin Zhang ◽  
...  
Keyword(s):  
Nucleus Pulposus ◽  
Dynamic Compression ◽  
Akt Pathway ◽  
Bioreactor Culture ◽  
Nucleus Pulposus Cells ◽  
Matrix Synthesis ◽  
Static Compression ◽  
Matrix Deposition ◽  
The Matrix

Background/Aims: An adequate matrix production of nucleus pulposus (NP) cells is an important tissue engineering-based strategy to regenerate degenerative discs. Here, we mainly aimed to investigate the effects and mechanism of mechanical compression (i.e., static compression vs. dynamic compression) on the matrix synthesis of three-dimensional (3D) cultured NP cells in vitro. Methods: Rat NP cells seeded on small intestinal submucosa (SIS) cryogel scaffolds were cultured in the chambers of a self-developed, mechanically active bioreactor for 10 days. Meanwhile, the NP cells were subjected to compression (static compression or dynamic compression at a 10% scaffold deformation) for 6 hours once per day. Unloaded NP cells were used as controls. The cellular phenotype and matrix biosynthesis of NP cells were investigated by real-time PCR and Western blotting assays. Lentivirus-mediated N-cadherin (N-CDH) knockdown and an inhibitor, LY294002, were used to further investigate the role of N-CDH and the PI3K/Akt pathway in this process. Results: Dynamic compression better maintained the expression of cell-specific markers (keratin-19, FOXF1 and PAX1) and matrix macromolecules (aggrecan and collagen II), as well as N-CDH expression and the activity of the PI3K/Akt pathway, in the 3D-cultured NP cells compared with those expression levels and activity in the cells grown under static compression. Further analysis showed that the N-CDH knockdown significantly down-regulated the expression of NP cell-specific markers and matrix macromolecules and inhibited the activation of the PI3K/Akt pathway under dynamic compression. However, inhibition of the PI3K/Akt pathway had no effects on N-CDH expression but down-regulated the expression of NP cell-specific markers and matrix macromolecules under dynamic compression. Conclusion: Dynamic compression increases the matrix synthesis of 3D-cultured NP cells compared with that of the cells under static compression, and the N-CDH-PI3K/Akt pathway is involved in this regulatory process. This study provides a promising strategy to promote the matrix deposition of tissue-engineered NP tissue in vitro prior to clinical transplantation.

scholarly journals Resveratrol inhibits IL-1β-mediated nucleus pulposus cell apoptosis through regulating the PI3K/Akt pathway

2019 ◽  
Vol 39 (3) ◽  
Author(s):  
Yanhai Jiang ◽  
Zhijie Xie ◽  
Jinying Yu ◽  
Lianqiang Fu
Keyword(s):  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
Cell Apoptosis ◽  
Inflammatory Cytokine ◽  
Culture Medium ◽  
Caspase 3 ◽  
Signal Transduction Pathway ◽  
Akt Pathway ◽  
Protective Effects ◽  
Apoptosis Ratio

Abstract Nucleus pulposus (NP) cell apoptosis is a classical cellular character during intervertebral disc degeneration (IDD). Previous studies have shown that inflammatory cytokine-induced NP cell apoptosis plays an important role in disc degeneration. The present study was aimed to investigate whether resveratrol can suppress IL-1β-mediated NP cell apoptosis and the potential signal transduction pathway. Experimental rat NP cells were treated with culture medium containing IL-1β (20 ng/ml) for 7 days. Control NP cells were cultured in the baseline medium. Resveratrol was added along with culture medium to investigate its effects. The inhibitor LY294002 was used to study the role of the PI3K/Akt pathway. NP cell apoptosis was reflected by the caspase-3 activity, cell apoptosis ratio, and expression of apoptosis-related molecules (Bcl-2, Bax, caspase-3, cleaved caspase-3, and cleaved PARP). Compared with the control NP cells, IL-1β significantly increased caspase-3 activity, NP cell apoptosis ratio and mRNA/protein expression of Bax, caspase-3, cleaved caspase-3 and cleaved PARP, but decreased mRNA expression of Bcl-2. However, resveratrol partly suppressed the effects of IL-1β on those cell apoptosis-related parameters. Further analysis showed that IL-1β significantly decreased activity of the PI3K/Akt pathway whereas resveratrol partly increased activity of the PI3K/Akt pathway in NP cells treated with IL-1β. Additionally, when the inhibitor LY294002 was added along with the resveratrol, its protective effects against IL-1β-induced NP cell apoptosis were attenuated. In conclusion, resveratrol suppresses IL-1β-mediated NP cell apoptosis through activating the PI3K/Akt pathway. Resveratrol may be an effective drug to attenuate inflammatory cytokine-induced disc degenerative changes.

scholarly journals Resveratrol attenuates high glucose-induced nucleus pulposus cell apoptosis and senescence through activating the ROS-mediated PI3K/Akt pathway

2018 ◽  
Vol 38 (2) ◽  
Author(s):  
Wenping Wang ◽  
Pei Li ◽  
Jiagang Xu ◽  
Xiangkun Wu ◽  
Zhiliang Guo ◽  
...  
Keyword(s):  
Nucleus Pulposus ◽  
High Glucose ◽  
Cell Apoptosis ◽  
Culture Medium ◽  
Nucleus Pulposus Cell ◽  
Akt Pathway ◽  
Control Group ◽  
Ros Generation ◽  
Protective Effects ◽  
Potential Mechanism

Background: Diabetes mellitus is closely correlated with disc degeneration. Nucleus pulposus (NP) cell apoptosis and senescence are typical cellular features within the degenerative disc. Resveratrol is a newly identified phytoalexin that has protective effects on cartilaginous tissue. Objective: To investigate the whether resveratrol can protect against high glucose-induced NP cell apoptosis and senescence, and the potential mechanism in this process. Methods: Rat NP cells were cultured in either 10% FBS culture medium (control group) or 10% FBS with a high glucose concentration (0.2 M, experiment group) for 3 days. Resveratrol or the combination of resveratrol and LY294002 was added into the culture medium of experiment group to investigate the effects of resveratrol and the PI3K/Akt pathway. Results: High glucose significantly promoted NP cell apoptosis and NP cell senescence compared with the control group. Resveratrol exhibited protective effects against high glucose-induced NP cell apoptosis and senescence. Further analysis showed that resveratrol suppressed reactive oxygen species (ROS) generation and increased the activity of the PI3K/Akt pathway under the high glucose condition. However, the LY294002 had no significant effects on ROS content in the resveratrol-treated high glucose group. Conclusion: Resveratrol can attenuate high glucose-induced NP cell apoptosis and senescence, and the activation of ROS-mediated PI3K/Akt pathway may be the potential mechanism in this process.

scholarly journals Knockout of miR-17-92 Cluster Protect Against Intervertebral Disc Degeneration by Inhibiting Apoptosis of Nucleus Pulposus Cells in Mice via the Activation of PI3K/Akt Pathway

2021 ◽  
Author(s):  
Yingjun Guo ◽  
Yang Meng ◽  
Hao Liu ◽  
Xiaofei Wang ◽  
Ying Hong ◽  
...  
Keyword(s):  
Intervertebral Disc ◽  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
Intervertebral Disc Degeneration ◽  
Mrna Level ◽  
Akt Pathway ◽  
Functional Genes ◽  
Control Group ◽  
Expression Levels ◽  
Protein Levels

Abstract Background: microRNA(miR)-17-92 cluster is involved in a variety of physiological and pathological processes, and the purpose of this study is to preliminarily explore the role of miR-17-92 cluster in disc degeneration and the corresponding mechanisms.Methods: Hematoxylin and Eosin (HE) and Safranin O Staining were used to evaluate the degeneration of intervertebral disc. qRT-PCR was applied to evaluate the mRNA level of miR-17-92 cluster and functional genes of nucleus pulposus (NP) tissues, whose protein level was evaluated with Western-blot. Terminal-Deoxynucleoitidyl Transferase Mediated Nick End Labeling (TUNEL) was used to evaluate the apoptotic level of nucleus pulposus cell (NPC).Results: The expression levels of members of the miR-17-92 cluster were significantly increased in the NP tissues from patients with intervertebral disc degeneration (IVDD). Furthermore, in the 3-months and 24-months miR-17-92-ccKO mice, the degree of IVDD was significantly lower than that of the control group. At the same time, we also detected the expression levels of related functional genes in the NP tissues of mice in two groups. The results showed that the mRNA and protein levels of Bax and Caspase-3 in the knockout group were significantly lower than those in the control group, and the mRNA and protein levels of Bcl-2 were significantly higher. The TUNEL results showed that the apoptosis level of NPCs in the 3-month knockout mice was significantly lower than that in the control group. Finally, the assessment of pathway-related protein levels showed that p-Ser473-Akt expression ratio in the nucleus pulposus of mice in the knockout group were significantly increased, suggesting that the PI3K/Akt pathway was activated after miR-17-92 cluster knockout.Conclusion: To sum up, miR-17-92 cluster does play an important regulating role in IVDD, and the results showed that miR-17-92 cluster could inhibiting NPCs apoptosis by activating PI3K/Akt pathway, eventually producing protective effect against IVDD.

Transplantation of Activated Nucleus Pulposus Cells after Cryopreservation: Efficacy Study in Canine Disc Degeneration Model

2014 ◽  
Vol 4 (1_suppl) ◽  
pp. s-0034-1376573-s-0034-1376573
Author(s):  
T. Nukaga ◽  
D. Sakai ◽  
A. Hiyama ◽  
T. Ishii ◽  
T. Nakai ◽  
...  
Keyword(s):  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
Efficacy Study ◽  
Nucleus Pulposus Cells
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