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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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ASC-CM suppresses ICAM-1 and E-selectin expression by HPAEC. HPAEC were treated with vehicle control (NoPr, black bars), NHDF-CM (DF-CM, grey bars), and hASC-CM (ASC-CM, white bars), and then stimulated with vehicle or 1 ng/ml TNFα (4 h). Harvested cells were analyzed for the surface expression of ICAM-1 and E-selectin. Experiments with independent treatment/stimulation were repeated in triplicate. T-test was used to assess the differences between analyzed groups. Exposure to hASC-CM lowers the percentage of ICAM-1-expressing quiescent HPAEC and reduces the level of E-selectin expression in TNFα-stimulated HPAEC.
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Fig10: ASC-CM suppresses ICAM-1 and E-selectin expression by HPAEC. HPAEC were treated with vehicle control (NoPr, black bars), NHDF-CM (DF-CM, grey bars), and hASC-CM (ASC-CM, white bars), and then stimulated with vehicle or 1 ng/ml TNFα (4 h). Harvested cells were analyzed for the surface expression of ICAM-1 and E-selectin. Experiments with independent treatment/stimulation were repeated in triplicate. T-test was used to assess the differences between analyzed groups. Exposure to hASC-CM lowers the percentage of ICAM-1-expressing quiescent HPAEC and reduces the level of E-selectin expression in TNFα-stimulated HPAEC.

Mentions: To ascertain whether hASC-CM anti-inflammatory effect is dependent on the suppression of endothelial expression of leucocyte receptors, we assessed the level of expression of VCAM, ICAM-1, and E-selectin in naïve and TNFα-stimulated HPAEC. Low percentage of unstimulated cells appeared positive for VCAM (data not shown) and E-selectin, whereas expression of ICAM-1 was detected in at least 70% cells (Figure 10). TNFα caused dramatic increase in the percentage of VCAM- (data not shown) and E-selectin-positive cells, also increasing ICAM-1 expression evident by the shift in ICAM-1 geo-mean value. HPAEC exposed to hASC-CM did not show significant difference in the expression of VCAM either in absence or presence of TNFα (data not shown). Percentage of ICAM-positive cells detected in the absence of TNFα was significantly suppressed by hASC-CM treatment; however, this effect was not detected in the presence of TNFα. On the contrary, E-selectin expression (evident by geo-mean) was significantly suppressed in the presence of TNFα.Figure 10


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)

ASC-CM suppresses ICAM-1 and E-selectin expression by HPAEC. HPAEC were treated with vehicle control (NoPr, black bars), NHDF-CM (DF-CM, grey bars), and hASC-CM (ASC-CM, white bars), and then stimulated with vehicle or 1 ng/ml TNFα (4 h). Harvested cells were analyzed for the surface expression of ICAM-1 and E-selectin. Experiments with independent treatment/stimulation were repeated in triplicate. T-test was used to assess the differences between analyzed groups. Exposure to hASC-CM lowers the percentage of ICAM-1-expressing quiescent HPAEC and reduces the level of E-selectin expression in TNFα-stimulated HPAEC.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig10: ASC-CM suppresses ICAM-1 and E-selectin expression by HPAEC. HPAEC were treated with vehicle control (NoPr, black bars), NHDF-CM (DF-CM, grey bars), and hASC-CM (ASC-CM, white bars), and then stimulated with vehicle or 1 ng/ml TNFα (4 h). Harvested cells were analyzed for the surface expression of ICAM-1 and E-selectin. Experiments with independent treatment/stimulation were repeated in triplicate. T-test was used to assess the differences between analyzed groups. Exposure to hASC-CM lowers the percentage of ICAM-1-expressing quiescent HPAEC and reduces the level of E-selectin expression in TNFα-stimulated HPAEC.
Mentions: To ascertain whether hASC-CM anti-inflammatory effect is dependent on the suppression of endothelial expression of leucocyte receptors, we assessed the level of expression of VCAM, ICAM-1, and E-selectin in naïve and TNFα-stimulated HPAEC. Low percentage of unstimulated cells appeared positive for VCAM (data not shown) and E-selectin, whereas expression of ICAM-1 was detected in at least 70% cells (Figure 10). TNFα caused dramatic increase in the percentage of VCAM- (data not shown) and E-selectin-positive cells, also increasing ICAM-1 expression evident by the shift in ICAM-1 geo-mean value. HPAEC exposed to hASC-CM did not show significant difference in the expression of VCAM either in absence or presence of TNFα (data not shown). Percentage of ICAM-positive cells detected in the absence of TNFα was significantly suppressed by hASC-CM treatment; however, this effect was not detected in the presence of TNFα. On the contrary, E-selectin expression (evident by geo-mean) was significantly suppressed in the presence of TNFα.Figure 10

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