[Sodium Ferulate Attenuates Oxidative Stress Induced Inflammation via Suppressing NALP3 Inflammasome and p38 MAPK Signal Pathway].

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-1ß) 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-1ß 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-1ß 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-1ß are reduced.

[Sodium Ferulate Attenuates Oxidative Stressvia Suppressing NALP3 Inflammasome and ERK Signal Pathway].

OBJECTIVES: Study the gene and protein expression of NACHT-PYD-containing protein 3 (NALP3) inflammasome and extracellular regulated protein kinase (ERK), the intervention effects of sodium ferulate (SF) in human lung epithelial cells A549 under oxidative stress, and to investigate the possible mechanism. METHODS: Human lung epithelial cells A549 cultured in vitro were divided into 6 groups, including control group,H2O2(200 umol/L) group, SF group (400 ug/mL), caspase-1 blockers (Z-VAD) group (Z-VAD 20 umol/L+H2O2200 umol/L), ERK blockers (PD98059) group (PD98059 50 umol/L+H2O2 200 umol/L), and SF+H2O2 group (SF 400 ug/mL+H2O2 200 umol/L). Fluorescent quantitative real-time PCR (qRT-PCR) was performed to detect the mRNA levels of caspase-1 and NALP3, the expression of caspase-1, NALP3, phosphorylated ERK p-ERK, ERK protein were evaluated by Western blot. The level of interleukin-1 ß (IL-1ß) were detected by ELISA. RESULTS: Compared with the control group,H2O2 not only increased the mRNA and protein expression levels of caspase-1 and NALP3 and the protein expression levels of p-ERK/ERK, but also enhanced the secretion of IL-1ß in human lung epithelial cells A549 (P<0.05),while SF group showed no statistic significance of those indicators above (P>0.05). The Z-VAD group, the PD98059 group and the SF+H2O2 group resisted the effects of H2O2 on A549 cells by decreasing the mRNA and protein expressions of caspase-1 and NALP3,and the protein expression of p-ERK/ERK, as well as reducing the secretion of IL-1ß(P<0.05),when compared with the H2O2 group. CONCLUSIONS: SF may reduce the expression of caspase-1, NALP3 and IL-1ß by inhibiting ERK, so as to reduce the inflammation caused by oxidative stress.