Acute Crit Care.  2019 May;34(2):133-140. 10.4266/acc.2019.00507.

The effects of BMS-470539 on lipopolysaccharide-induced acute lung injury

• Jang EA
• Kim JY
• Tin TD
• Song JA
• Lee SH
• Kwak SH

• KMID: 2449367
• DOI: http://doi.org/10.4266/acc.2019.00507

Abstract

BACKGROUND
Overactivation of inflammatory cells, including macrophages and neutrophils, is associated with acute lung injury. BMS-470539 is a selective agonist of melanocortin 1 receptor, which triggers the inhibition of proinflammatory responses, suppressing neutrophil infiltration and protecting tissue. This study evaluated the effects of BMS-470539 on lipopolysaccharide-induced acute lung injury in a mouse model.
METHODS
Mice received a subcutaneous injection of saline or BMS-470539 (18.47 mg/kg) 1 hour before an intratracheal injection of saline or lipopolysaccharide (20 µg). Mice were sacrificed to analyze the severity of pulmonary edema (lung wet-to-dry weight [W/D] ratio) and inflammatory responses (level of leukocytes, polymorphonuclear neutrophils [PMNs] and tumor necrosis factor alpha [TNF-α] in bronchoalveolar lavage fluid [BALF]), and neutrophil infiltration (myeloperoxidase activity). TNF-α activation was also measured in neutrophils from bone marrow. Survival was investigated in a second-hit sepsis mouse model.
RESULTS
BMS-470539 improved sepsis-induced pulmonary edema, as demonstrated by a decreased W/D ratio (5.76%±0.83% to 3.81%±0.86%, P<0.05). The inflammatory response also improved, as shown by decreased levels of leukocytes (551±116 to 357±86×10²/mm³, P<0.05), PMNs (51.52%±16.23% to 18.41%±7.25%, P<0.01), and TNF-α (550±338 to 128±52 pg/ml, P<0.01) in the BALF. BMS-470539 also improved the inflammatory response, as shown by TNF-α levels (850±158 to 423±59 pg/ml, P<0.01) in neutrophils. BMS-470539 downregulated neutrophil infiltration in the lung (myeloperoxidase: 654±98 to 218±89 U/g, P<0.001). Lastly, BMS improved the survival rate (0% to 70%, P<0.01) in a mice multiple organ failure model.
CONCLUSIONS
BMS-470539 improved lipopolysaccharide-induced acute lung injury and mortality in mice by affecting the inflammatory response.

Keyword

acute lung injury; cytokines; lipopolysaccharides; melanocortin 1 receptor

MeSH Terms

Acute Lung Injury*
Animals
Bone Marrow
Bronchoalveolar Lavage Fluid
Cytokines
Injections, Subcutaneous
Leukocytes
Lipopolysaccharides
Lung
Macrophages
Mice
Mortality
Multiple Organ Failure
Neutrophil Infiltration
Neutrophils
Pulmonary Edema
Receptor, Melanocortin, Type 1
Sepsis
Survival Rate
Tumor Necrosis Factor-alpha
Cytokines
Lipopolysaccharides
Receptor, Melanocortin, Type 1
Tumor Necrosis Factor-alpha

Figure

• 

  Figure 1. (A, B) Animal model of LPS-induced acute lung injury. Mice were randomly assigned to one of four groups: mice receiving subcutaneous (SC) saline (0.1 ml) and intratracheal (IT) saline (40 μl) (control model [CON] group, n=10); mice receiving SC saline and IT lipopolysaccharide (LPS) 20 μg (40 μl) (LPS group, n=10); mice receiving SC BMS-470539 18.47 mg/kg (0.1 ml) and IT saline (BMS group, n=10); mice receiving SC BMS-470539 18.47 mg/kg (0.1 ml) and IT LPS (BMS+LPS group, n=10). Mice were initially anesthetized with sevoflurane. The SC injection of saline or BMS performed 1 hour prior to the IT injection of saline or LPS. Animals were euthanized to collect the BALF and lung tissues at 18 hours after the IT administration of saline or LPS. PBS: phosphate-buffered saline; BALF: bronchoalveolar lavage fluid.

• 

  Figure 2. The effects of BMS-470539 on cytokine expression (TNF-α) in LPS-stimulated neutrophils. Neutrophils from mouse bone marrow were incubated with the indicated concentration of BMS470539 with or without LPS (100 ng/ml) for 4 hours, and then TNF-α levels were measured in the culture supernatants by ELISA. Each bar represents the mean±standard error of the mean. Data are representative of three separate experiments. TNF-α: tumor necrosis factor alpha; CON: control model; LPS: lipopolysaccharide; ELISA: enzyme-linked immunosorbent assay. aP<0.01, vs. CON; bP<0.01, vs. LPS.

• 

  Figure 3. The effects of BMS-470539 on MPO content in the right lung. MPO was measured in the right lungs of the control mice (CON) and mice treated with BMS-470539 (18.47 mg/kg) alone (BMS), lipopolysaccharide (20 μg) alone (LPS), or BMS-470539 (18.47 mg/kg) before lipopolysaccharide (20 μg) (BMS+LPS). The data are from five mice in each group. Each bar represents the mean±standard error of the mean. MPO: myeloperoxidase. aP<0.001, vs. CON; bP<0.001 vs. LPS.

• 

  Figure 4. The effects of BMS-470539 on the wet-to-dry weight (W/D) ratio in the left lung. The W/D ratio was measured in the left lungs of the control mice (CON) and mice treated with BMS-470539 (18.47 mg/kg) alone (BMS), lipopolysaccharide (20 μg) alone (LPS), or BMS-470539 (18.47 mg/kg) before lipopolysaccharide (20 μg) (BMS+LPS). The data are from five mice in each group. Each bar represents the mean±standard error of the mean. aP<0.01, vs. CON; bP<0.05, vs. LPS.

• 

  Figure 5. The effects of BMS-470539 on mortality in a multiple organ failure (MOF) animal model. Survival (10 mice per group) was monitored for 14 days after establishment of the MOF model. Mice were intraperitoneally administered Escherichia coli 055:B5 lipopolysaccharide (LPS; 5 mg/kg body weight) to induce the initial septic insult, and at 24 hours post-LPS administration, a medium-grade cecal ligation and puncture (25-gauge needle, double puncture) procedure was performed to create the MOF model. To determine the effects of BMS-470539, all mice that received cecal ligation and puncture were divided into two groups: (1) the MOF group, and (2) the BMS group: at 24 hours post-LPS administration, the mice received an intraperitoneal injection of BMS-470539 (18.47 mg/kg body weight). In the sham control model, the mice were opened abdominally to perform the same surgical procedure, but their cecum was neither ligated nor punctured. All models were monitored twice daily for 14 days or until mortality.

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