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Conditioned media from adipose stromal cells limit lipopolysaccharide-induced lung injury, endothelial hyperpermeability and apoptosis.

Lu H, Poirier C, Cook T, Traktuev DO, Merfeld-Clauss S, Lease B, Petrache I, March KL, Bogatcheva NV - J Transl Med (2015)

Bottom Line: ASC-CM markedly reduced LPS-induced histopathologic changes of lung, protein extravasation into BALF, and suppressed the secretion of proinflammatory cytokines TNFα and IL6.ASC-CM exposure reduced the percentage of endothelial cells expressing ICAM-1, and suppressed TNFα-induced expression of E-selectin and cleavage of caspase-3.ASC-CM reduced the endothelial level of pro-apoptotic protein Bim, but did not affect the level of Bcl-2, Bad, or Bad phosphorylation.

View Article: PubMed Central - PubMed

Affiliation: Division of Cardiology, Indiana University, Indianapolis, IN, USA. honlu@iu.edu.

ABSTRACT

Background: Acute Respiratory Distress Syndrome (ARDS) is a condition that contributes to morbidity and mortality of critically ill patients. We investigated whether factors secreted by adipose stromal cells (ASC) into conditioned media (ASC-CM) will effectively decrease lung injury in the model of lipopolysaccharide (LPS)-induced ARDS.

Methods: To assess the effect of ASC-CM on ARDS indices, intravenous delivery of ASC and ASC-CM to C57Bl/6 mice was carried out 4 h after LPS oropharyngeal aspiration; Evans Blue Dye (EBD) was injected intravenously 1 h prior to animal sacrifice (48 h post-LPS). Lungs were either fixed for histopathology, or used to extract bronchoalveolar lavage fluid (BALF) or EBD. To assess the effect of ASC-CM on endothelial barrier function and apoptosis, human pulmonary artery endothelial cells were treated with ASC-CM for 48-72 h.

Results: ASC-CM markedly reduced LPS-induced histopathologic changes of lung, protein extravasation into BALF, and suppressed the secretion of proinflammatory cytokines TNFα and IL6. White Blood Cells (WBC) from BALF of LPS-challenged mice receiving ASC-CM had decreased reactive oxygen species (ROS) generation compared to WBC from LPS-challenged mice receiving control media injection. Treatment of pulmonary endothelial monolayers with ASC-CM significantly suppressed H2O2-induced leakage of FITC dextran and changes in transendothelial resistance, as well as gap formation in endothelial monolayer. ASC-CM exposure reduced the percentage of endothelial cells expressing ICAM-1, and suppressed TNFα-induced expression of E-selectin and cleavage of caspase-3. ASC-CM reduced the endothelial level of pro-apoptotic protein Bim, but did not affect the level of Bcl-2, Bad, or Bad phosphorylation.

Conclusions: Factors secreted by ASC efficiently reduce ARDS indices, endothelial barrier hyperpermeability, and activation of pro-inflammatory and pro-apoptotic pathways in endothelium.

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Related in: MedlinePlus

Surface marker expression of mASC (A) and hASC (B). IC-isotype controls, AS- antigen-specific antibodies. As expected, the majority of mASCs expressed the surface antigen of mesenchymal stem cells Sca-1, and was negative for the hematopoietic stem cell marker (CD45) and endothelial marker (CD31). The majority of hASC expressed stromal markers CD13, CD73, CD90, and CD105, and was negative for CD31, CD45.
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Fig1: Surface marker expression of mASC (A) and hASC (B). IC-isotype controls, AS- antigen-specific antibodies. As expected, the majority of mASCs expressed the surface antigen of mesenchymal stem cells Sca-1, and was negative for the hematopoietic stem cell marker (CD45) and endothelial marker (CD31). The majority of hASC expressed stromal markers CD13, CD73, CD90, and CD105, and was negative for CD31, CD45.

Mentions: Both murine and human ASC used in our study were produced by propagation of the cell fraction adherent to plastic under standard culture conditions. mASC and hASC demonstrated typical size, spindle shape, and morphology in culture consistent with adipose stem cells. mASC as well as hASC were evaluated with respect to their surface marker phenotype at passage 3. mASC were positive for Sca-1, one of the markers routinely used to characterize mesenchymal stem cells of murine origin [30], and negative for the endothelial cell marker CD31 and the marker of cells of hematopoietic origin CD45 (Figure 1A). Similarly, analysis of hASC at passage 3 demonstrated positive staining for the stromal markers CD13, CD73, CD90 and CD105 and negative staining for CD31 and CD45 (Figure 1B).Figure 1


Conditioned media from adipose stromal cells limit lipopolysaccharide-induced lung injury, endothelial hyperpermeability and apoptosis.

Lu H, Poirier C, Cook T, Traktuev DO, Merfeld-Clauss S, Lease B, Petrache I, March KL, Bogatcheva NV - J Transl Med (2015)

Surface marker expression of mASC (A) and hASC (B). IC-isotype controls, AS- antigen-specific antibodies. As expected, the majority of mASCs expressed the surface antigen of mesenchymal stem cells Sca-1, and was negative for the hematopoietic stem cell marker (CD45) and endothelial marker (CD31). The majority of hASC expressed stromal markers CD13, CD73, CD90, and CD105, and was negative for CD31, CD45.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4358867&req=5

Fig1: Surface marker expression of mASC (A) and hASC (B). IC-isotype controls, AS- antigen-specific antibodies. As expected, the majority of mASCs expressed the surface antigen of mesenchymal stem cells Sca-1, and was negative for the hematopoietic stem cell marker (CD45) and endothelial marker (CD31). The majority of hASC expressed stromal markers CD13, CD73, CD90, and CD105, and was negative for CD31, CD45.
Mentions: Both murine and human ASC used in our study were produced by propagation of the cell fraction adherent to plastic under standard culture conditions. mASC and hASC demonstrated typical size, spindle shape, and morphology in culture consistent with adipose stem cells. mASC as well as hASC were evaluated with respect to their surface marker phenotype at passage 3. mASC were positive for Sca-1, one of the markers routinely used to characterize mesenchymal stem cells of murine origin [30], and negative for the endothelial cell marker CD31 and the marker of cells of hematopoietic origin CD45 (Figure 1A). Similarly, analysis of hASC at passage 3 demonstrated positive staining for the stromal markers CD13, CD73, CD90 and CD105 and negative staining for CD31 and CD45 (Figure 1B).Figure 1

Bottom Line: ASC-CM markedly reduced LPS-induced histopathologic changes of lung, protein extravasation into BALF, and suppressed the secretion of proinflammatory cytokines TNFα and IL6.ASC-CM exposure reduced the percentage of endothelial cells expressing ICAM-1, and suppressed TNFα-induced expression of E-selectin and cleavage of caspase-3.ASC-CM reduced the endothelial level of pro-apoptotic protein Bim, but did not affect the level of Bcl-2, Bad, or Bad phosphorylation.

View Article: PubMed Central - PubMed

Affiliation: Division of Cardiology, Indiana University, Indianapolis, IN, USA. honlu@iu.edu.

ABSTRACT

Background: Acute Respiratory Distress Syndrome (ARDS) is a condition that contributes to morbidity and mortality of critically ill patients. We investigated whether factors secreted by adipose stromal cells (ASC) into conditioned media (ASC-CM) will effectively decrease lung injury in the model of lipopolysaccharide (LPS)-induced ARDS.

Methods: To assess the effect of ASC-CM on ARDS indices, intravenous delivery of ASC and ASC-CM to C57Bl/6 mice was carried out 4 h after LPS oropharyngeal aspiration; Evans Blue Dye (EBD) was injected intravenously 1 h prior to animal sacrifice (48 h post-LPS). Lungs were either fixed for histopathology, or used to extract bronchoalveolar lavage fluid (BALF) or EBD. To assess the effect of ASC-CM on endothelial barrier function and apoptosis, human pulmonary artery endothelial cells were treated with ASC-CM for 48-72 h.

Results: ASC-CM markedly reduced LPS-induced histopathologic changes of lung, protein extravasation into BALF, and suppressed the secretion of proinflammatory cytokines TNFα and IL6. White Blood Cells (WBC) from BALF of LPS-challenged mice receiving ASC-CM had decreased reactive oxygen species (ROS) generation compared to WBC from LPS-challenged mice receiving control media injection. Treatment of pulmonary endothelial monolayers with ASC-CM significantly suppressed H2O2-induced leakage of FITC dextran and changes in transendothelial resistance, as well as gap formation in endothelial monolayer. ASC-CM exposure reduced the percentage of endothelial cells expressing ICAM-1, and suppressed TNFα-induced expression of E-selectin and cleavage of caspase-3. ASC-CM reduced the endothelial level of pro-apoptotic protein Bim, but did not affect the level of Bcl-2, Bad, or Bad phosphorylation.

Conclusions: Factors secreted by ASC efficiently reduce ARDS indices, endothelial barrier hyperpermeability, and activation of pro-inflammatory and pro-apoptotic pathways in endothelium.

Show MeSH
Related in: MedlinePlus