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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

ASC-CM suppresses activation of pro-apoptotic pathways in HPAEC. HPAEC were treated with vehicle control, NHDF-CM, and hASC-CM, and then stimulated with indicated concentration of TNFα (4 h). Cell lysates were analyzed with antibodies to cleaved caspase 3, Bad, phospho-Bad, Bim, and Bcl2. β-actin staining was used as loading control.
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Fig11: ASC-CM suppresses activation of pro-apoptotic pathways in HPAEC. HPAEC were treated with vehicle control, NHDF-CM, and hASC-CM, and then stimulated with indicated concentration of TNFα (4 h). Cell lysates were analyzed with antibodies to cleaved caspase 3, Bad, phospho-Bad, Bim, and Bcl2. β-actin staining was used as loading control.

Mentions: To elucidate whether anti-inflammatory effects of hASC-CM are mediated by the suppression of pro-apoptotic changes in endothelium, we assessed the level of caspase-3 cleavage in response to TNFα (Figure 11). TNFα induced cleavage and activation of caspase-3 evident at 4 h. HPAEC pretreated with hASC-CM showed a marked reduction of cleaved caspase-3 level in response to TNFα. As caspases activation is known to be regulated by pro-apoptotic and anti-apoptotic members of Bcl2 family [31], we next investigated whether levels of these proteins are affected by hASC secreted factors. Levels of anti-apoptotic protein Bcl2 and pro-apoptotic protein Bad did not change in HPAEC preconditioned with hASC-CM. We did not observe consistent increase in Bad phosphorylation (evident of Bad deactivation) in response to hASC-CM pretreatment. However, exposure to hASC-secreted factors markedly reduced the level of pro-apoptotic protein Bim.Figure 11


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 activation of pro-apoptotic pathways in HPAEC. HPAEC were treated with vehicle control, NHDF-CM, and hASC-CM, and then stimulated with indicated concentration of TNFα (4 h). Cell lysates were analyzed with antibodies to cleaved caspase 3, Bad, phospho-Bad, Bim, and Bcl2. β-actin staining was used as loading control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig11: ASC-CM suppresses activation of pro-apoptotic pathways in HPAEC. HPAEC were treated with vehicle control, NHDF-CM, and hASC-CM, and then stimulated with indicated concentration of TNFα (4 h). Cell lysates were analyzed with antibodies to cleaved caspase 3, Bad, phospho-Bad, Bim, and Bcl2. β-actin staining was used as loading control.
Mentions: To elucidate whether anti-inflammatory effects of hASC-CM are mediated by the suppression of pro-apoptotic changes in endothelium, we assessed the level of caspase-3 cleavage in response to TNFα (Figure 11). TNFα induced cleavage and activation of caspase-3 evident at 4 h. HPAEC pretreated with hASC-CM showed a marked reduction of cleaved caspase-3 level in response to TNFα. As caspases activation is known to be regulated by pro-apoptotic and anti-apoptotic members of Bcl2 family [31], we next investigated whether levels of these proteins are affected by hASC secreted factors. Levels of anti-apoptotic protein Bcl2 and pro-apoptotic protein Bad did not change in HPAEC preconditioned with hASC-CM. We did not observe consistent increase in Bad phosphorylation (evident of Bad deactivation) in response to hASC-CM pretreatment. However, exposure to hASC-secreted factors markedly reduced the level of pro-apoptotic protein Bim.Figure 11

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