Epigallocatechin 3-gallate suppresses interleukin-1β-induced inflammatory responses in intervertebral disc cells in vitro and reduces radiculopathic pain in rats

Abstract

Intervertebral disc (IVD) disease, which is characterised by age-related changes in the adult disc, is the most common cause of disc failure and low back pain. The purpose of this study was to analyse the potential of the biologically active polyphenol epigallocatechin 3-gallate (EGCG) for the treatment of painful IVD disease by identifying and explaining its anti-inflammatory and anti-catabolic activity. Human IVD cells were isolated from patients undergoing surgery due to degenerative disc disease (n = 34) and cultured in 2D or 3D. An inflammatory response was activated by IL-1, EGCG was added, and the expression/activity of inflammatory mediators and pathways was measured by qRT-PCR, western blotting, ELISA, immunofluorescence and transcription factor assay. The small molecule inhibitor SB203580 was used to investigate the involvement of the p38 pathway in the observed effects. The analgesic properties of EGCG were analysed by the von Frey filament test in Sprague-Dawley rats (n = 60). EGCG significantly inhibited the expression of pro-inflammatory mediators and matrix metalloproteinases in vitro, as well as radiculopathic pain in vivo, most probably by modulation of the activity of IRAK-1 and its downstream effectors p38, JNK and NF-B. AbstractIntervertebral disc (IVD) disease, which is characterised by age-related changes in the adult disc, is the most common cause of disc failure and low back pain. The purpose of this study was to analyse the potential of the biologically active polyphenol epigallocatechin 3-gallate (EGCG) for the treatment of painful IVD disease by identifying and explaining its anti-inflammatory and anti-catabolic activity. Human IVD cells were isolated from patients undergoing surgery due to degenerative disc disease (n = 34) and cultured in 2D or 3D. An inflammatory response was activated by IL-1β, EGCG was added, and the expression/activity of inflammatory mediators and pathways was measured by qRT-PCR, western blotting, ELISA, immunofluorescence and transcription factor assay. The small molecule inhibitor SB203580 was used to investigate the involvement of the p38 pathway in the observed effects. The analgesic properties of EGCG were analysed by the von Frey filament test in SpragueDawley rats (n = 60). EGCG significantly inhibited the expression of pro-inflammatory mediators and matrix metalloproteinases in vitro, as well as radiculopathic pain in vivo, most probably by modulation of the activity of IRAK-1 and its downstream effectors p38, JNK and NF-κB.

(-)-Epigallocatechin-3-Gallate Inhibits Arsenic-Induced Inflammation and Apoptosis through Suppression of Oxidative Stress in Mice

“…Accumulating evidence has revealed that EGCG, as a potent antioxidative agent, can inhibit inflammation and enhance the survival of disc cells in oxidative stress [31]. The anti-inflammatory effects of EGCG on radiculopathic pain have also been investigated in vitro and may benefit from its inhibitory effect on the overexpression of pro-inflammatory mediators [13, 32]. Given the inhibition of immune cell-derived cellular functions by arsenic exposure and the antioxidative and anti-inflammatory properties of EGCG in antagonizing increased secretion of inflammatory mediators, simultaneous exposure to EGCG has been found to be beneficial in increasing arsenic mobilization and assisting the recovery of altered biochemical levels during chemical therapy.…”

Chondrogenic Priming at Reduced Cell Density Enhances Cartilage Adhesion of Equine Allogeneic MSCs - a Loading Sensitive Phenomenon in an Organ Culture Study with 180 Explants

“…As indicated in Fig. 1, MSCs were cultured in normal expansion medium or induction medium for 6, 24 or 72 hours of culture before being lysed in 1 ml of Trizol (Invitrogen) and RNA was isolated by standard chloroform phase separation (250 µl, Sigma-Aldrich) as previously described [25]. RNA was quantified on the Nanodrop Lite (Fisher Scientific) before reverse transcribing 1 µg of RNA into cDNA (10 minutes at 25°C, 120 minutes at 37°C), using the TaqMan Reverse Transcription Reagents Kit (Life Technologies).…”

The effect of serotonin–noradrenaline reuptake inhibitor duloxetine on the intervertebral disk-related radiculopathy in rats

“…Sensitivity to non-noxious mechanical stimuli was tested by previously established von Frey test [16,22,29,35,36,40,42,45,52]. Behavioral tests were performed by physician unaware of the experimental groupings.…”