Molecular hydrogen protects against oxidative stress‑induced RAW 264.7 macrophage cells through the activation of Nrf2 and inhibition of MAPK signaling pathway

Abstract

Background: Oxidative stress is involved in the development of many inflammatory, metabolic and aging diseases. Objective: In this study we investigated, the protective effects of H2 on RAW 264.7 macrophage cell against LPS-and H2O2-induced oxidative stress by the inhibition of MAPK pathway and also activate the Nrf2 pathway. Results: Our results showed H2 increased the macrophage cell proliferation and generated ROS and NO against LPS stimulation to exert an active immune response. Similarly, H2 protected the macrophage cell from H2O2-induced oxidative stress. H2 reduced the LPS-and H2O2-induced inflammatory cytokine production and intracellular calcium influxes. H2 inhibited the LPS-and H2O2-induced phosphorylation of MAPK pathway and its downstream signaling molecules. Furthermore, H2 protected the macrophage cell from mitochondrial apoptosis. H2 increased Nrf2 protein expression indicating its strong anti-oxidative effects against oxidative stress. Conclusion: Collectively, our results indicate the strong antioxidant role of H2 against LPS-and H2O2-induced oxidative stress on macrophage cells by activating the Nrf2 pathway and inhibiting the MAPK-signaling pathway.

Study details

Article type

Original Research

Disease / injury model

Oxidative stress and inflammation (LPS- and H2O2-induced) in RAW 264.7 macrophage cells

Dose comparison

The study compared the effects of different concentrations of H2 (0.3, 0.4, 0.5 mM) and different exposure times (30, 60, 120 min) on oxidative stress and inflammatory markers in RAW 264.7 cells.

Dose-dependent effect

Yes

Outcome

Positive

Findings

Molecular hydrogen (H2) protects RAW 264.7 macrophage cells from LPS- and H2O2-induced oxidative stress by activating the Nrf2 pathway and inhibiting the MAPK signaling pathway, resulting in reduced inflammatory cytokine production, decreased intracellular calcium influx, increased antioxidant enzyme activity, and reduced apoptosis.