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In Vitro Stimulation and Visualization of Extracellular Trap Release in Differentiated Human Monocyte-derived Macrophages
Published on: November 1, 2019
Hydrogen peroxide stimulates macrophages and monocytes to actively release HMGB1
Daolin Tang1, Yongzhong Shi, Rui Kang
1Department of Pathophysiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China.
Abstract:
High mobility group box 1 (HMGB1) can be actively secreted by macrophages/monocytes in response to exogenous and endogenous inflammatory stimuli (such as bacterial endotoxin, TNF-alpha, IL-1, and IFN-gamma) or passively released by necrotic cells and mediates innate and adaptive inflammatory responses to infection and injury. Here, we demonstrated that a reactive oxygen species, hydrogen peroxide (H(2)O(2)), induces active and passive HMGB1 release from macrophage and monocyte cultures in a time- and dose-dependent manner. At nontoxic doses (e.g., 0.0125-0.125 mM), H(2)O(2) induced HMGB1 cytoplasmic translocation and active release within 3-24 h. At higher concentrations (e.g., 0.25 mM), however, H(2)O(2) exhibited cytotoxicity to macrophage and monocyte cell cultures and consequently, triggered active and passive HMGB1 release. In addition, H(2)O(2) stimulated potential interaction of HMGB1 with a nuclear export factor, chromosome region maintenance (CRM1), in macrophage/monocyte cultures. Inhibitors specific for the JNK (SP600125) and MEK (PD98059), but not p38 MAPK (SB203580), abrogated H(2)O(2)-induced, active HMGB1 release. Together, these data establish an important role for oxidative stress in inducing active HMGB1 release, potentially through a MAPK- and CRM1-dependent mechanism.
Insights
Oxidative stress from hydrogen peroxide (H2O2) triggers the release of High Mobility Group Box 1 (HMGB1) from immune cells. This release involves cellular mechanisms dependent on MAPK and CRM1 signaling pathways.
Area of Science:
- Immunology
- Cell Biology
- Oxidative Stress Research
Background:
- High Mobility Group Box 1 (HMGB1) is a crucial mediator of inflammatory responses, secreted by immune cells or released from necrotic cells.
- HMGB1 plays a significant role in both innate and adaptive immunity during infection and injury.
- Understanding the triggers for HMGB1 release is vital for comprehending inflammatory processes.
Purpose of the Study:
- To investigate the effect of hydrogen peroxide (H2O2), a reactive oxygen species, on HMGB1 release from macrophages and monocytes.
- To elucidate the mechanisms underlying H2O2-induced HMGB1 release, including cellular translocation and signaling pathways.
Main Methods:
- Macrophage and monocyte cultures were treated with varying concentrations of H2O2.
- HMGB1 release, cytoplasmic translocation, and cytotoxicity were assessed over time.
- The involvement of specific signaling pathways (MAPK, JNK, MEK, p38) and nuclear export factor CRM1 was investigated using inhibitors.
Main Results:
- H2O2 induced both active and passive HMGB1 release in a time- and dose-dependent manner.
- Non-toxic H2O2 concentrations promoted HMGB1 cytoplasmic translocation and active release.
- Higher H2O2 concentrations caused cytotoxicity, leading to both active and passive HMGB1 release.
- H2O2 stimulated the interaction between HMGB1 and the nuclear export factor CRM1.
- JNK and MEK pathway inhibitors, but not p38 MAPK inhibitors, abrogated H2O2-induced active HMGB1 release.
Conclusions:
- Oxidative stress, specifically H2O2, plays a significant role in inducing active HMGB1 release from macrophages and monocytes.
- The H2O2-induced HMGB1 release mechanism appears to involve mitogen-activated protein kinase (MAPK) and chromosome region maintenance (CRM1) signaling.
- These findings highlight a novel link between oxidative stress and inflammatory mediator release, with potential implications for inflammatory diseases.
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