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Lack of inhibitory effects of the anti-fibrotic drug imatinib on endothelial cell functions in vitro and in vivo.

Venalis P, Maurer B, Akhmetshina A, Busch N, Dees C, Stürzl M, Zwerina J, Jüngel A, Gay S, Schett G, Distler O, Distler JH - J. Cell. Mol. Med. (2009)

Bottom Line: No reduction in proliferation or metabolic activity of endothelial cells was observed.Imatinib did not affect migration of endothelial cells and did not reduce the formation of capillary tubes.Imatinib does not inhibit activation, viability, proliferation, migration or tube forming of endothelial cells in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department for Internal Medicine 3 and Institute for Clinical Immunology, Friedrich-Alexander-University Erlangen-Nuremberg, Erlangen, Germany.

ABSTRACT
Systemic sclerosis (SSc) is a systemic autoimmune disease that is characterized by microangiopathy with progressive loss of capillaries and tissue fibrosis. Imatinib exerts potent anti-fibrotic effects and is currently evaluated in clinical trials. The aim of the present study was to exclude that the anti-fibrotic effects of imatinib are complicated by inhibitory effects on endothelial cell functions, which might augment vascular disease in SSc. Endothelial cells and mice were treated with pharmacologically relevant concentrations of imatinib. The expression of markers of vascular activation was assessed with real-time PCR. Proliferation was analysed with the cell counting experiments and the MTT assay. Apoptosis was quantified with caspase 3 assays, annexin V in vitro and with TUNEL staining in vivo. Migration was studied with scratch and transwell assays. Tube forming was investigated with the matrigel assay. Imatinib did not alter the expression of markers of vascular activation. Imatinib did not increase the percentage of annexin V positive cells or the activity of caspase 3. No reduction in proliferation or metabolic activity of endothelial cells was observed. Imatinib did not affect migration of endothelial cells and did not reduce the formation of capillary tubes. Consistent with the in vitro data, no difference in the number of apoptotic endothelial cells was observed in vivo in mice treated with imatinib. Imatinib does not inhibit activation, viability, proliferation, migration or tube forming of endothelial cells in vitro and in vivo. Thus, treatment with imatinib might not augment further endothelial cell damage in SSc.

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Imatinib does not induce apoptosis, reduce the metabolic activity or decrease proliferation of endothelial cells. (A and B) Incubation with imatinib in concentrations from 0.001 to 1.0 μg/ml for 24 hrs did not increase the activity of caspase 3 (n= 3) (A) or the number of annexin V positive HMEC-1 cells (n= 9) (B). (C) Incubation with imatinib in pharmacologically relevant concentrations for 48 hrs did not reduce the metabolic activity of HMEC-1 cells compared with cells incubated with medium only (n= 5). In contrast, incubation with 50% DMSO profoundly reduced the metabolic activity. (D) Imatinib did not reduce proliferation of HMEC-1 cells as analysed by direct cell counting (n= 6).
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fig01: Imatinib does not induce apoptosis, reduce the metabolic activity or decrease proliferation of endothelial cells. (A and B) Incubation with imatinib in concentrations from 0.001 to 1.0 μg/ml for 24 hrs did not increase the activity of caspase 3 (n= 3) (A) or the number of annexin V positive HMEC-1 cells (n= 9) (B). (C) Incubation with imatinib in pharmacologically relevant concentrations for 48 hrs did not reduce the metabolic activity of HMEC-1 cells compared with cells incubated with medium only (n= 5). In contrast, incubation with 50% DMSO profoundly reduced the metabolic activity. (D) Imatinib did not reduce proliferation of HMEC-1 cells as analysed by direct cell counting (n= 6).

Mentions: The effect of imatinib on apoptosis of microvascular endothelial cells was analysed by quantification of caspase 3 activity and staining for annexin V. After incubation with imatinib mesylate in concentrations from 0.001 to 1.0 μg/ml, no changes of the activity of caspase 3 were detectable (Fig. 1A). Incubation of HMEC-1 cells with increasing concentrations of imatinib did also not increase the number of annexin V positive, apoptotic cells (Fig. 1B). To investigate whether imatinib augments pre-existing endothelial cell damage, HMEC-1 cells were serum starved and then incubated with imatinib. As under normal culture conditions, imatinib did neither induce activation of caspase 3 nor increase the number of apoptotic cells.


Lack of inhibitory effects of the anti-fibrotic drug imatinib on endothelial cell functions in vitro and in vivo.

Venalis P, Maurer B, Akhmetshina A, Busch N, Dees C, Stürzl M, Zwerina J, Jüngel A, Gay S, Schett G, Distler O, Distler JH - J. Cell. Mol. Med. (2009)

Imatinib does not induce apoptosis, reduce the metabolic activity or decrease proliferation of endothelial cells. (A and B) Incubation with imatinib in concentrations from 0.001 to 1.0 μg/ml for 24 hrs did not increase the activity of caspase 3 (n= 3) (A) or the number of annexin V positive HMEC-1 cells (n= 9) (B). (C) Incubation with imatinib in pharmacologically relevant concentrations for 48 hrs did not reduce the metabolic activity of HMEC-1 cells compared with cells incubated with medium only (n= 5). In contrast, incubation with 50% DMSO profoundly reduced the metabolic activity. (D) Imatinib did not reduce proliferation of HMEC-1 cells as analysed by direct cell counting (n= 6).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4496125&req=5

fig01: Imatinib does not induce apoptosis, reduce the metabolic activity or decrease proliferation of endothelial cells. (A and B) Incubation with imatinib in concentrations from 0.001 to 1.0 μg/ml for 24 hrs did not increase the activity of caspase 3 (n= 3) (A) or the number of annexin V positive HMEC-1 cells (n= 9) (B). (C) Incubation with imatinib in pharmacologically relevant concentrations for 48 hrs did not reduce the metabolic activity of HMEC-1 cells compared with cells incubated with medium only (n= 5). In contrast, incubation with 50% DMSO profoundly reduced the metabolic activity. (D) Imatinib did not reduce proliferation of HMEC-1 cells as analysed by direct cell counting (n= 6).
Mentions: The effect of imatinib on apoptosis of microvascular endothelial cells was analysed by quantification of caspase 3 activity and staining for annexin V. After incubation with imatinib mesylate in concentrations from 0.001 to 1.0 μg/ml, no changes of the activity of caspase 3 were detectable (Fig. 1A). Incubation of HMEC-1 cells with increasing concentrations of imatinib did also not increase the number of annexin V positive, apoptotic cells (Fig. 1B). To investigate whether imatinib augments pre-existing endothelial cell damage, HMEC-1 cells were serum starved and then incubated with imatinib. As under normal culture conditions, imatinib did neither induce activation of caspase 3 nor increase the number of apoptotic cells.

Bottom Line: No reduction in proliferation or metabolic activity of endothelial cells was observed.Imatinib did not affect migration of endothelial cells and did not reduce the formation of capillary tubes.Imatinib does not inhibit activation, viability, proliferation, migration or tube forming of endothelial cells in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department for Internal Medicine 3 and Institute for Clinical Immunology, Friedrich-Alexander-University Erlangen-Nuremberg, Erlangen, Germany.

ABSTRACT
Systemic sclerosis (SSc) is a systemic autoimmune disease that is characterized by microangiopathy with progressive loss of capillaries and tissue fibrosis. Imatinib exerts potent anti-fibrotic effects and is currently evaluated in clinical trials. The aim of the present study was to exclude that the anti-fibrotic effects of imatinib are complicated by inhibitory effects on endothelial cell functions, which might augment vascular disease in SSc. Endothelial cells and mice were treated with pharmacologically relevant concentrations of imatinib. The expression of markers of vascular activation was assessed with real-time PCR. Proliferation was analysed with the cell counting experiments and the MTT assay. Apoptosis was quantified with caspase 3 assays, annexin V in vitro and with TUNEL staining in vivo. Migration was studied with scratch and transwell assays. Tube forming was investigated with the matrigel assay. Imatinib did not alter the expression of markers of vascular activation. Imatinib did not increase the percentage of annexin V positive cells or the activity of caspase 3. No reduction in proliferation or metabolic activity of endothelial cells was observed. Imatinib did not affect migration of endothelial cells and did not reduce the formation of capillary tubes. Consistent with the in vitro data, no difference in the number of apoptotic endothelial cells was observed in vivo in mice treated with imatinib. Imatinib does not inhibit activation, viability, proliferation, migration or tube forming of endothelial cells in vitro and in vivo. Thus, treatment with imatinib might not augment further endothelial cell damage in SSc.

Show MeSH
Related in: MedlinePlus