Acute Crit Care.  2018 Aug;33(3):154-161. 10.4266/acc.2018.00619.

Anti-inflammatory Role of Mesenchymal Stem Cells in an Acute Lung Injury Mouse Model

Affiliations
  • 1Department of Pulmonary and Critical Care Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea. sbhong@amc.seoul.kr
  • 2Department of Emergency Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea.
  • 3Asan Institute for Life Sciences, Seoul, Korea.

Abstract

BACKGROUND
Mesenchymal stem cells (MSCs) attenuate injury in various lung injury models through paracrine effects. We hypothesized that intratracheal transplantation of allogenic MSCs could attenuate lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice, mediated by anti-inflammatory responses.
METHODS
Six-week-old male mice were randomized to either the control or the ALI group. ALI was induced by intratracheal LPS instillation. Four hours after LPS instillation, MSCs or phosphate-buffered saline was randomly intratracheally administered. Neutrophil count and protein concentration in bronchoalveolar lavage fluid (BALF); lung histology; levels of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and macrophage inflammatory protein-2; and the expression of proliferation cell nuclear antigen (PCNA), caspase-3, and caspase-9 were evaluated at 48 hours after injury.
RESULTS
Treatment with MSCs attenuated lung injury in ALI mice by decreasing protein level and neutrophil recruitment into the BALF and improving the histologic change. MSCs also decreased the protein levels of proinflammatory cytokines including IL-1β, IL-6, and TNF-α, but had little effect on the protein expression of PCNA, caspase-3, and caspase-9.
CONCLUSIONS
Intratracheal injection of bone marrow-derived allogenic MSCs attenuates LPSinduced ALI via immunomodulatory effects.

Keyword

acute lung injury; cytokines; immunomodulation; inflammation; lipopolysaccharides

MeSH Terms

Acute Lung Injury*
Animals
Bronchoalveolar Lavage Fluid
Caspase 3
Caspase 9
Cytokines
Humans
Immunomodulation
Inflammation
Interleukin-6
Interleukins
Lipopolysaccharides
Lung
Lung Injury
Macrophages
Male
Mesenchymal Stromal Cells*
Mice*
Neutrophil Infiltration
Neutrophils
Proliferating Cell Nuclear Antigen
Tumor Necrosis Factor-alpha
Caspase 3
Caspase 9
Cytokines
Interleukin-6
Interleukins
Lipopolysaccharides
Proliferating Cell Nuclear Antigen
Tumor Necrosis Factor-alpha

Figure

  • Figure 1. Mesenchymal stem cells attenuate lung injury in lipopolysaccharide-induced acute lung injury. (A) Total cell counts of bronchoalveolar lavage fluid (BALF). (B) Percentage of neutrophils in BALF. (C) Total protein level in BALF. (D) Myeloperoxidase activity in serum. CTL: control group; MSC: mesenchymal stem cell injected group; LPS: lipopolysaccharide-induced acute lung injury group; LPS/MSC: MSC injection following LPS induced acute lung injury group. *P<0.05 compared to the CTL group; **P<0.05 compared to the LPS group.

  • Figure 2. Light microscopy findings of the lung. (A, B) In the control (CTL) group, the normal parenchymal architecture, including the alveolar septa, alveolar lumen, and capillaries were well preserved. The infiltration of inflammatory cells was not observed. (C, D) In the mesenchymal stem cell (MSC) injected group, the normal pulmonary architecture was well preserved similar to that in the CTL group. (E, F) In the lipopolysaccharide (LPS)-induced acute lung injury group, the pulmonary architecture, including the alveolar septa and alveolar lumen were not preserved. Perivascular edema (asterisk) and intrapulmonary hemorrhage (arrow heads) were observed. The infiltration of inflammatory cells, perivascular cuffing (black arrows), and floating of phagocytes in the alveolar lumen (white arrows) were observed. (G, H) In the MSC injection following LPS induced acute lung injury (LPS/MSC) group, the alveolar septa and alveolar lumen were relatively well preserved. Intrapulmonary hemorrhage and perivascular cuffing was less prominent compared to those of the lungs in the LPS group. The infiltration of inflammatory cells was not observed. (H&E; magnification, A, C, E, G: ×200; B, D, F, H: ×400).

  • Figure 3. Lung injury scores. CTL: control group; MSC: mesenchymal stem cell injected group; LPS: lipopolysaccharide-induced acute lung injury group; LPS/MSC: MSC injection following LPS induced acute lung injury group. *P<0.05, compared to the CTL group; **P<0.05, compared to the LPS group.

  • Figure 4. Mesenchymal stem cells attenuate lipopolysaccharide-induced acute lung injury by anti-inflammatory effects. (A) Tumor necrosis factor (TNF)-α, (B) interleukin (IL)-1β, (C) IL-6, and (D) macrophage inflammatory protein (MIP)-2 in bronchoalveolar lavage fluid (BALF). CTL: control group; MSC: mesenchymal stem cell injected group; LPS: lipopolysaccharide-induced acute lung injury group; LPS/MSC: MSC injection following LPS induced acute lung injury group. *P<0.05, compared to the CTL group; **P<0.05, compared to the LPS group.

  • Figure 5. The effect of mesenchymal stem cells on proliferation and cell death. (A) Western blot of proliferation cell nuclear antigen (PCNA), caspase-9 and cleaved-caspase-9, caspase-3, and β-actin. (B) Relative protein level (PCNA/β-actin ratio). CTL: control group; MSC: mesenchymal stem cell injected group; LPS: lipopolysaccharide-induced acute lung injury group; LPS/MSC: MSC injection following LPS induced acute lung injury group.


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