Pharmacological NRF2 activation inhibits <i>Mcp-1</i> transcript accumulation.

<p>(A) Real time PCR analysis for <i>Mcp-1</i> in healthy donor monocytes treated with CDDO-Me(bardoxolone methyl) (50nM). Values are relative to unstimulated, vehicle treated cells at indicated Hours Post-LPS Stimulation (HPS). Results are representative of 8 independent donors. (B) Real time PCR analysis for <i>Il-6</i> in healthy donor monocytes treated with CDDO-Me(bardoxolone methyl) (50nM). Values are relative to unstimulated, vehicle treated cells at indicated HPS. Results are representative of 6 independent donors. (C) Real time PCR analysis for <i>Il-10</i> in healthy donor monocytes treated with CDDO-Me(bardoxolone methyl) (50nM). Values are relative to unstimulated, vehicle treated cells at indicated Hours Post-LPS Stimulation (HPS). Results are representative of 6 independent donors. (D) Real time PCR analysis for <i>Nqo-1</i> in healthy donor monocytes treated with CDDO-Me(bardoxolone methyl) (50nM). Values are relative to unstimulated, vehicle treated cells at indicated HPS. Results are representative of 4 independent donors. (E) Bar graph representing the combination of CDDO-Me(bardoxolone methyl)-altered <i>Mcp-1</i> transcript accumulation in 8 donors. (F) Bar graph representing the combination of CDDO-Me(bardoxolone methyl)-altered <i>Il-6</i> transcript accumulation in 6 donors. (G) Bar graph representing the combination of CDDO-Me(bardoxolone methyl)-altered <i>Il-10</i> transcript accumulation in 6 donors. (H) Bar graph representing the combination of CDDO-Me(bardoxolone methyl)-altered <i>Nqo-1</i> transcript accumulation in 4 donors. Error bars represent SEM. * = <i>p</i> < .05. ** = <i>p</i> < .01. *** = <i>p</i> < .001. **** = <i>p</i> < .0001.</p

Systemic IL-6 and MCP-1 are elevated in patients with large burn injuries and organ dysfunction.

<p>(A) Scatter plot analysis of systemic IL-6 levels between mild (<15% TBSA, Black circles) and moderate/severe (≥15% TBSA, Red squares) patients at 0–48 HPA. (B) Scatter plot analysis of systemic MCP-1 levels between mild (<15% TBSA, Black circles) and moderate/severe (≥15% TBSA, Red squares) patients at 0–48 HPA. (C) Scatter plot analysis of systemic IL-6 levels between burn patients that suffer pulmonary distress(0–24 HPA, <357 SpO₂/FiO₂ ratio) and patients that did not (>357 SpO₂/FiO₂ ratio). (D) Scatter plot analysis of systemic MCP-1 levels between burn patients that suffer pulmonary distress(0–24 HPA, <357 SpO₂/FiO₂ ratio) and patients that did not (>357 SpO₂/FiO₂ ratio). (E) Line plot representing longitudinal analysis of systemic IL-6 accumulation from 0–48 HPA and 72–144 HPA timepoints. (F) Line plot representing longitudinal analysis of systemic MCP-1 accumulation from 0–48 HPA and 72–144 HPA timepoints. Error bars represent ± SEM. T-tests were performed to determine statistical significance for (A-D). **** represents <i>P</i> < .0001, *** represent <i>P</i> < .001, * represent <i>P</i> < .05.</p

Multiple NRF2 agonists reduce MCP-1 production in enriched monocyte populations.

<p>(A) ELISA analysis for MCP-1, IL-6, and IL-10 in healthy donor monocytes after treatment with CDDO-Me(bardoxolone methyl) (50nM) at indicated Hours Post-LPS Stimulation (HPS). Results are representative of 5 independent donors. (B) Bar graph depicting the percentage of CDDO-Me(bardoxolone methyl)-altered cytokine production for MCP-1, IL-6, and IL-10 at 16 and 24 HPS. Relative values represent the combination of 5 independent sets of healthy donor monocytes. (C) Immunoblot analysis demonstrating the kinetics of NQO1 expression after treatment with CDDO-Me(bardoxolone methyl) (50nM). Results are representative of 5 independent donors. (D) ELISA analysis for MCP-1, IL-6, and IL-10 in healthy donor monocytes after treatment with Compound 7 (1 μM) at 16 HPS. Results are representative of 4 independent donors. (E) Bar graph depicting the percentage of Compound 7-altered cytokine production for MCP-1, IL-6, and IL-10 at 16 HPS. Relative values represent the combination of 4 independent sets of healthy donor monocytes. (F) Immunoblot analysis demonstrating NQO1 expression after treatment with Compound 7 (1 μM) at 16 HPS. Results are representative of 4 independent donors. Error bars represent SEM. ** = <i>p</i> < .01. *** = <i>p</i> < .001. **** = <i>p</i> < .0001.</p

NRF2 activation attenuated IL-6-induced MCP-1 production.

<p>(A) ELISA analysis for MCP-1 in healthy donor monocytes after treatment with CDDO-Me(bardoxolone methyl) (50nM) at indicated Hours Post-IL-6 Stimulation (HPS). Results are representative of 4 independent donors. (B) Bar graph depicting the percentage of CDDO-Me(bardoxolone methyl)-altered MCP-1 production at 16 HPS. Relative values represent the combination of 4 independent sets of healthy donor monocytes. (C) Immunoblot analysis demonstrating NQO1 expression after treatment with CDDO-Me(bardoxolone methyl) (50nM) at 16 HPA. Results are representative of 2 independent donors. (D) ELISA analysis for MCP-1 in healthy donor monocytes after treatment with Compound 7 (1 μM) at 16 HPS. Results are representative of 2 independent donors. Error bars represent SEM. * = <i>p</i> < .05. ** = <i>p</i> < .01. *** = <i>p</i> < .001. **** = <i>p</i> < .0001.</p

Pharmacological NRF2 activation reduced MCP-1 production in burn patients immune cells.

<p>(A) Line plot representing the CDDO-Me(bardoxolone methyl)(50 nM)-mediated changes in MCP-1 production across 30 patients (0–72 HPA). Red squares and lines indicate moderate/severe burn group (≥15% TBSA) while black circles and lines represent mild burn group (<15% TBSA). (B) Line plot representing the CDDO-Me(bardoxolone methyl)(50 nM)-mediated changes in IL-6 production across 29 patients (0–72 HPA). Red squares and lines indicate moderate/severe burn group (≥15% TBSA) while black circles and lines represent mild burn group (<15% TBSA). (C) Line plot representing the CDDO-Me(bardoxolone methyl) (50 nM)-mediated changes in IL-10 production across 23 patients (0–72 HPA). Red squares and lines indicate moderate/severe burn group (≥15% TBSA) while black circles and lines represent mild burn group (<15% TBSA). (D) Bar graph depicting the percentage of CDDO-Me(bardoxolone methyl)-altered cytokine production for MCP-1, IL-6, and IL-10. Relative values were combined across the patient cohort (N = 30 for MCP-1, N = 29 for IL-6, N = 23 for IL-10). Error bars represent ± SEM.</p