Sodium Ferulate Attenuates Oxidative Stress Induced Inflammation via Suppressing NALP3 Inflammasome and p38 MAPK Signal Pathway
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Abstract
Objective To investigate the changes of NACHT-PYD-containing protein 3(NALP3) inflammasome and p38 mitogen-activated protein kinase (MARK), and the interventional mechanism of sodium ferulate (SF) in mouse embryonic fibroblasts (NIH-3T3 cells) under oxidative stress. Methods NIH-3T3 cells cultured in vitro were divided into 6 groups, including control group, H2O2 stress group (H2O2 200 μmol/L), SF group (SF 400 μg/mL), antioxidant group (H2O2 200 μmol/L+NAC 5 mmol/L), p38 MAPK blockers group (H2O2 200 μmol/L+SB203580 5 μmol/L), H2O2+SF group (H2O2 200 μmol/L+SF 400 μg/mL). The mRNA expression levels of NALP3, Caspase-1 and p38α were evaluated by fluorescent quantitative real-time PCR (qRT-PCR) at 24 h after cell culture; Western blot was performed to detect the expressions of NALP3, p-p38 to p38 protein (the activation of p38 MAPK signaling pathway expressed by the ratio of p-p38 to p38) at 48 h after cell culture; The levels of interleukin-1beta (IL-1β) in different groups were detected by ELISA at 2 h after cell culture. Results Compared with control group, H2O2 could not only increase the expression levels of NALP3, Caspase-1, p38α mRNAs and NALP3, p-p38/p38 proteins but also the secretion of IL-1β in NIH-3T3 cells when compared with the control group (P<0.05); Antioxidant NAC, p38 MAPK blockers and H2O2+SF group could partly resisted the effects of H2O2 on NIH-3T3 cells, that decreased the level of mRNA expressions of NALP3, Caspase-1, p38α and protein expressions of NALP3 and p-p38/p38, and reduced the secretion of IL-1β when compared to the H2O2 stress group (P<0.05); However, H2O2+NAC group, SB203580 group and H2O2+SF group showed no statistical difference of those indicators (P>0.05). Conclusion The mechanism of sodium ferulate attenuated oxidative stress induced inflammation may be through blunting p38 MAPK signal pathway, the expression of NALP3 inflammasome, Caspase-1 and the secretion of IL-1β are reduced.
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