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Kinetics of lung tissue factor expression and procoagulant activity in bleomycin induced acute lung injury.

Ma L, Shaver CM, Grove BS, Mitchell DB, Wickersham NE, Carnahan RH, Cooper TL, Brake BE, Ware LB, Bastarache JA - Clin Transl Med (2015)

Bottom Line: BAL protein and lung wet-to-dry weight ratio increased significantly by day 3.Changes in permeability and procoagulant activity preceded inflammatory cell influx which was maximal at day 6 while whole lung TF protein peaked along with inflammation.These data demonstrate that cytokine upregulation is the earliest response to bleomycin administration, followed by increased lung permeability, upregulation of TF, and recruitment of inflammatory cells.

View Article: PubMed Central - PubMed

Affiliation: Division of Emergency Intensive Care Unit, the Second Hospital of Lanzhou University, Lanzhou, China, mali1105@126.com.

ABSTRACT

Background: Activation of coagulation by expression of tissue factor (TF) in the airspace is a hallmark of acute lung injury (ALI) but the timing of TF activation in relationship to increases in lung permeability and inflammation are unknown.

Methods: To test the hypothesis that TF is upregulated early in the course of acute bleomycin lung injury and precedes increased permeability and inflammation we studied the early course of bleomycin-induced ALI in mice. Mice were treated with 0.04U intratracheal bleomycin or vehicle control and bronchoalveolar lavage (BAL) and lung tissue were collected daily for 7 days. Whole lung TF mRNA was determined by QT-PCR. TF protein was assessed by ELISA and immunostaining. BAL procoagulant activity was measured by BAL clot time and thrombin-antithrombin complexes. Inflammation was assessed by BAL cell count, differentials and CXCL1/KC concentration. Lung permeability was assessed by BAL protein and lung wet to dry weight ratio.

Results: Expression of CXCL1 occurred by day 1. BAL protein and lung wet-to-dry weight ratio increased significantly by day 3. TF mRNA and BAL procoagulant activity peaked on day 4 while whole lung TF protein peaked on day 6. Changes in permeability and procoagulant activity preceded inflammatory cell influx which was maximal at day 6 while whole lung TF protein peaked along with inflammation.

Conclusion: These data demonstrate that cytokine upregulation is the earliest response to bleomycin administration, followed by increased lung permeability, upregulation of TF, and recruitment of inflammatory cells.

No MeSH data available.


Related in: MedlinePlus

BAL procoagulant activity over time. TF procoagulant activity was measured by BAL clot time (Panel a) and BAL thrombin-antithrombin complexes (Panel b) over time in response to IT bleomycin. Clot time is expressed as percent of the saline-treated animals on the same day. N = 5-6 per group. *p = 0.024 versus day 1 **p < 0.001 and §p = 0.028 versus PBS control by t test on each day
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Fig3: BAL procoagulant activity over time. TF procoagulant activity was measured by BAL clot time (Panel a) and BAL thrombin-antithrombin complexes (Panel b) over time in response to IT bleomycin. Clot time is expressed as percent of the saline-treated animals on the same day. N = 5-6 per group. *p = 0.024 versus day 1 **p < 0.001 and §p = 0.028 versus PBS control by t test on each day

Mentions: In order to define the time course of TF regulation we measured TF mRNA and protein (Fig. 1), cellular localization by immunostaining (Fig. 2) and activity (Fig. 3) over time in response to IT bleomycin treatment. In the bleomycin treated group, TF mRNA increased slightly starting at day 2 with peaks at day 4 (Fig. 1a). TF protein in whole lung homogenates as measured by ELISA (normalized to total protein) increased daily and peaked at day 6 (Fig. 1b). Increased TF protein expression in the lung epithelium was evident by day 1 by TF immunostaining (Fig. 2). Staining became more patchy and intense on days 3 and 5 and persisted on day 7. These increases in TF mRNA and protein were paralleled by increased TF procoagulant activity in BAL fluid (Fig. 3a) which also peaked at day 4. Consistent with TF procoagulant activity and generation of thrombin, thrombin anti-thrombin complexes started to increase at day 3 and peak on day 6 (Fig. 3b).Fig. 1


Kinetics of lung tissue factor expression and procoagulant activity in bleomycin induced acute lung injury.

Ma L, Shaver CM, Grove BS, Mitchell DB, Wickersham NE, Carnahan RH, Cooper TL, Brake BE, Ware LB, Bastarache JA - Clin Transl Med (2015)

BAL procoagulant activity over time. TF procoagulant activity was measured by BAL clot time (Panel a) and BAL thrombin-antithrombin complexes (Panel b) over time in response to IT bleomycin. Clot time is expressed as percent of the saline-treated animals on the same day. N = 5-6 per group. *p = 0.024 versus day 1 **p < 0.001 and §p = 0.028 versus PBS control by t test on each day
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495096&req=5

Fig3: BAL procoagulant activity over time. TF procoagulant activity was measured by BAL clot time (Panel a) and BAL thrombin-antithrombin complexes (Panel b) over time in response to IT bleomycin. Clot time is expressed as percent of the saline-treated animals on the same day. N = 5-6 per group. *p = 0.024 versus day 1 **p < 0.001 and §p = 0.028 versus PBS control by t test on each day
Mentions: In order to define the time course of TF regulation we measured TF mRNA and protein (Fig. 1), cellular localization by immunostaining (Fig. 2) and activity (Fig. 3) over time in response to IT bleomycin treatment. In the bleomycin treated group, TF mRNA increased slightly starting at day 2 with peaks at day 4 (Fig. 1a). TF protein in whole lung homogenates as measured by ELISA (normalized to total protein) increased daily and peaked at day 6 (Fig. 1b). Increased TF protein expression in the lung epithelium was evident by day 1 by TF immunostaining (Fig. 2). Staining became more patchy and intense on days 3 and 5 and persisted on day 7. These increases in TF mRNA and protein were paralleled by increased TF procoagulant activity in BAL fluid (Fig. 3a) which also peaked at day 4. Consistent with TF procoagulant activity and generation of thrombin, thrombin anti-thrombin complexes started to increase at day 3 and peak on day 6 (Fig. 3b).Fig. 1

Bottom Line: BAL protein and lung wet-to-dry weight ratio increased significantly by day 3.Changes in permeability and procoagulant activity preceded inflammatory cell influx which was maximal at day 6 while whole lung TF protein peaked along with inflammation.These data demonstrate that cytokine upregulation is the earliest response to bleomycin administration, followed by increased lung permeability, upregulation of TF, and recruitment of inflammatory cells.

View Article: PubMed Central - PubMed

Affiliation: Division of Emergency Intensive Care Unit, the Second Hospital of Lanzhou University, Lanzhou, China, mali1105@126.com.

ABSTRACT

Background: Activation of coagulation by expression of tissue factor (TF) in the airspace is a hallmark of acute lung injury (ALI) but the timing of TF activation in relationship to increases in lung permeability and inflammation are unknown.

Methods: To test the hypothesis that TF is upregulated early in the course of acute bleomycin lung injury and precedes increased permeability and inflammation we studied the early course of bleomycin-induced ALI in mice. Mice were treated with 0.04U intratracheal bleomycin or vehicle control and bronchoalveolar lavage (BAL) and lung tissue were collected daily for 7 days. Whole lung TF mRNA was determined by QT-PCR. TF protein was assessed by ELISA and immunostaining. BAL procoagulant activity was measured by BAL clot time and thrombin-antithrombin complexes. Inflammation was assessed by BAL cell count, differentials and CXCL1/KC concentration. Lung permeability was assessed by BAL protein and lung wet to dry weight ratio.

Results: Expression of CXCL1 occurred by day 1. BAL protein and lung wet-to-dry weight ratio increased significantly by day 3. TF mRNA and BAL procoagulant activity peaked on day 4 while whole lung TF protein peaked on day 6. Changes in permeability and procoagulant activity preceded inflammatory cell influx which was maximal at day 6 while whole lung TF protein peaked along with inflammation.

Conclusion: These data demonstrate that cytokine upregulation is the earliest response to bleomycin administration, followed by increased lung permeability, upregulation of TF, and recruitment of inflammatory cells.

No MeSH data available.


Related in: MedlinePlus