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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

mASC and mASC-CM effect on the level of anti-inflammatory cytokine IL10 and pro-angiogenic factor VEGF. IL10 was detected in BALF of LPS-challenged mice injected with mASC (A), but not mASC-CM (data not shown). LPS-induced increase in VEGF level was reduced in response to mASC-CM injection (C), but not mASC injection (B). One-way Anova with Tukey post-hoc was used to assess the significance of differences between the groups; number of animals is indicated.
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Fig7: mASC and mASC-CM effect on the level of anti-inflammatory cytokine IL10 and pro-angiogenic factor VEGF. IL10 was detected in BALF of LPS-challenged mice injected with mASC (A), but not mASC-CM (data not shown). LPS-induced increase in VEGF level was reduced in response to mASC-CM injection (C), but not mASC injection (B). One-way Anova with Tukey post-hoc was used to assess the significance of differences between the groups; number of animals is indicated.

Mentions: To determine the effect of ASC and ASC-CM on inflammation progression/resolution, we assessed the level of the pro-inflammatory cytokines TNFα, IL6, and MIP-2, and anti-inflammatory IL10 in BALF. LPS caused marked increase in the level of TNFα, IL6 and MIP-2 (Figure 6A-F). Forty-eight hours after cell or conditioned media injection, LPS-induced increase in TNFα and IL6 levels was suppressed by both agents, whereas the increase in MIP-2 level was not affected significantly. The level of anti-inflammatory IL10 remained below detection in control mice as well as mice challenged with LPS. Significant increase in BALF IL10 content was observed in response to ASC (Figure 7A), but not ASC-CM (data not shown). Interestingly, the LPS-induced level of VEGF in BALF was non-significantly increased in ASC-treated mice, but markedly suppressed in ASC-CM-treated mice (Figure 7B-C).Figure 6


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)

mASC and mASC-CM effect on the level of anti-inflammatory cytokine IL10 and pro-angiogenic factor VEGF. IL10 was detected in BALF of LPS-challenged mice injected with mASC (A), but not mASC-CM (data not shown). LPS-induced increase in VEGF level was reduced in response to mASC-CM injection (C), but not mASC injection (B). One-way Anova with Tukey post-hoc was used to assess the significance of differences between the groups; number of animals is indicated.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig7: mASC and mASC-CM effect on the level of anti-inflammatory cytokine IL10 and pro-angiogenic factor VEGF. IL10 was detected in BALF of LPS-challenged mice injected with mASC (A), but not mASC-CM (data not shown). LPS-induced increase in VEGF level was reduced in response to mASC-CM injection (C), but not mASC injection (B). One-way Anova with Tukey post-hoc was used to assess the significance of differences between the groups; number of animals is indicated.
Mentions: To determine the effect of ASC and ASC-CM on inflammation progression/resolution, we assessed the level of the pro-inflammatory cytokines TNFα, IL6, and MIP-2, and anti-inflammatory IL10 in BALF. LPS caused marked increase in the level of TNFα, IL6 and MIP-2 (Figure 6A-F). Forty-eight hours after cell or conditioned media injection, LPS-induced increase in TNFα and IL6 levels was suppressed by both agents, whereas the increase in MIP-2 level was not affected significantly. The level of anti-inflammatory IL10 remained below detection in control mice as well as mice challenged with LPS. Significant increase in BALF IL10 content was observed in response to ASC (Figure 7A), but not ASC-CM (data not shown). Interestingly, the LPS-induced level of VEGF in BALF was non-significantly increased in ASC-treated mice, but markedly suppressed in ASC-CM-treated mice (Figure 7B-C).Figure 6

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