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Heme scavenging reduces pulmonary endoplasmic reticulum stress, fibrosis, and emphysema
Saurabh Aggarwal, … , Mark T. Dransfield, Sadis Matalon
Saurabh Aggarwal, … , Mark T. Dransfield, Sadis Matalon
Published November 2, 2018
Citation Information: JCI Insight. 2018;3(21):e120694. https://doi.org/10.1172/jci.insight.120694.
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Research Article Pulmonology

Heme scavenging reduces pulmonary endoplasmic reticulum stress, fibrosis, and emphysema

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Abstract

Pulmonary fibrosis and emphysema are irreversible chronic events after inhalation injury. However, the mechanism(s) involved in their development remain poorly understood. Higher levels of plasma and lung heme have been recorded in acute lung injury associated with several insults. Here, we provide the molecular basis for heme-induced chronic lung injury. We found elevated plasma heme in chronic obstructive pulmonary disease (COPD) (GOLD stage 4) patients and also in a ferret model of COPD secondary to chronic cigarette smoke inhalation. Next, we developed a rodent model of chronic lung injury, where we exposed C57BL/6 mice to the halogen gas, bromine (Br2) (400 ppm, 30 minutes), and returned them to room air resulting in combined airway fibrosis and emphysematous phenotype, as indicated by high collagen deposition in the peribronchial spaces, increased lung hydroxyproline concentrations, and alveolar septal damage. These mice also had elevated pulmonary endoplasmic reticulum (ER) stress as seen in COPD patients; the pharmacological or genetic diminution of ER stress in mice attenuated Br2-induced lung changes. Finally, treating mice with the heme-scavenging protein, hemopexin, reduced plasma heme, ER stress, airway fibrosis, and emphysema. This is the first study to our knowledge to report elevated heme in COPD patients and establishes heme scavenging as a potential therapy after inhalation injury.

Authors

Saurabh Aggarwal, Israr Ahmad, Adam Lam, Matthew A. Carlisle, Changzhao Li, J. Michael Wells, S. Vamsee Raju, Mohammad Athar, Steven M. Rowe, Mark T. Dransfield, Sadis Matalon

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Figure 9

Hemopexin attenuates lung injury, airway fibrosis, and lung emphysema.

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Hemopexin attenuates lung injury, airway fibrosis, and lung emphysema.
M...
Male C57BL/6 mice were exposed to air or Br2 gas (400 ppm, 30 minutes) and then returned to room air. Some Br2-exposed mice were given an intraperitoneal injection of purified human hemopexin (Hx) (4 μg/g BW) 1 hour or 5 days after Br2 exposure. All air-exposed mice and some Br2-exposed mice received saline injection as an appropriate control. Fourteen days after Br2 exposure, mouse BALF protein levels (n = 5–9) (A) and total cell count (n = 5–7) (B) were elevated in saline-treated mice but were significantly lower in Hx-treated mice. Hx-treated mice had decreased lung deposition of collagen on Masson’s trichrome staining (n = 5–8) (C) and lower lung hydroxyproline levels (n = 5–8) (D) compared with saline-treated mice, 14 days after Br2 inhalation. Assessment of mouse lung pressure-volume (P-V) curves demonstrated that Br2 exposure increased lung volumes, as indicated by the shifting up and left of PV curve in the saline-treated mice but not in the Hx-treated mice (n = 5–10) (F). Hematoxylin and eosin (H&E) staining of lungs showed that Hx prevented Br2-induced alveolar septa damage (n = 5–8) (C) and reduced alveolar Lm (n = 5–8). Scale bars 100 µm. (E). In addition, Hx lowered plasma elastase levels (n = 14–16) (G) and BALF elastase activity (n = 10–13) (H) in Br2-exposed mice. The Kaplan-Meier curve demonstrated that Hx reduced mortality after Br2 exposure (n = 42 for Br2 + saline; n = 20 for Br2 + Hx [1 hour after]; n = 17 for Br2 + Hx [5 days after]) (I). *P < 0.05 versus air + saline and †P < 0.05 versus Br2 + saline by 1-way ANOVA followed by Tukey’s post hoc test. PV curves were analyzed by 2-way ANOVA with Bonferroni’s post hoc test. Overall survival was analyzed by the Kaplan-Meier method. Differences in survival were tested for statistical significance by the log-rank test.

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