Protective effects of ginsenoside Rg3 on TNF-α-induced human nucleus pulposus cells through inhibiting NF-κB signaling pathway

This work aims to evaluate the effect of ginsenoside Rg3 on the apoptosis, proliferation, extracellular matrix (ECM) metabolism and oxidative stress-induced damage of human nucleus pulposus cells (NPCs) induced by TNF-α. The human NPCs were divided into Control, TNF-α, TNF-α + low Rg3, TNF-α + mediu...

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Bibliographic Details
Published in:Life sciences (1973) 2019-01, Vol.216, p.1-9
Main Authors: Chen, Jiang, Liu, Gen-Zhe, Sun, Qi, Zhang, Fan, Liu, Chu-yin, Yuan, Lin, Zhao, Xue-Qian, Wang, Yong-Jun, Jia, Yu-Song
Format: Article
Language:English
Subjects:
Aggrecan
Annexin V
Apoptosis
Apoptosis - drug effects
BAX protein
Bcl-2 protein
Caspase
Caspase-3
Cell cycle
Cell Proliferation - drug effects
Cells, Cultured
Cholecystokinin
Control
Enzyme-linked immunosorbent assay
Extracellular matrix
Extracellular Matrix - metabolism
Flow Cytometry
Gene expression
Genes
Ginsenosides
Ginsenosides - administration & dosage
Ginsenosides - pharmacology
Glutathione
Glutathione peroxidase
Humans
Intervertebral discs
Malondialdehyde
Metabolism
NF-kappa B - metabolism
NF-κB
NF-κB protein
Nuclei (cytology)
Nucleus pulposus
Nucleus Pulposus - drug effects
Nucleus Pulposus - pathology
Nucleus pulposus cells
Oxidative metabolism
Oxidative stress
Oxidative Stress - drug effects
Peroxidase
Proteins
Reactive oxygen species
Reactive Oxygen Species - metabolism
Rg3
Signal transduction
Signal Transduction - drug effects
Signaling
Superoxide dismutase
TNF-α
Transcription Factor RelA - metabolism
Tumor Necrosis Factor-alpha - administration & dosage
Up-Regulation
Western blotting
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Summary:This work aims to evaluate the effect of ginsenoside Rg3 on the apoptosis, proliferation, extracellular matrix (ECM) metabolism and oxidative stress-induced damage of human nucleus pulposus cells (NPCs) induced by TNF-α. The human NPCs were divided into Control, TNF-α, TNF-α + low Rg3, TNF-α + medium Rg3 and TNF-α + high Rg3 groups. Annexin V-FITC/PI, CCK-8 and flow cytometry were used to detect the apoptosis, proliferation, and cell cycle of NPCs, respectively. The expressions of ECM-related molecules were determined by qRT-PCR, ELISA and Western blotting. NF-κB p65 pathway and apoptosis-related proteins were evaluated by Western blotting, and the production of reactive oxygen species (ROS) was detected by DCFH-DA assay. Compared with Control group, NPCs in the TNF-α group had elevated proportion of apoptotic cells with up-regulation of Bax and Caspase-3 and down-regulation of Bcl-2. Besides, TNF-α inhibited proliferation and arrested cell cycle at G1 of NPCs. Moreover, human NPCs induced by TNF-α presented the increase in the expressions of ECM degrading genes (MMP3 and ADAMTS5), the content of ROS and malondialdehyde (MDA), and the expression of NF-κB/p65 in nucleus, but showed the decrease in the expression of ECM synthesis genes (Aggrecan and COL2A1) and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX). However, NPCs treated by both TNF-α and Rg3 demonstrated a certain degree of reversal in the above indexes, which became increasingly evident with the up-regulation of Rg3 concentration. Ginsenoside Rg3 may exert the effect of attenuating TNF-α-induced NPCs impairment via blocking the NF-κB signaling pathway.
ISSN:0024-3205
1879-0631
DOI:10.1016/j.lfs.2018.11.022