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Transforming growth factor-β1 requires NADPH oxidase 4 for angiogenesis in vitro and in vivo.

Peshavariya HM, Chan EC, Liu GS, Jiang F, Dusting GJ - J. Cell. Mol. Med. (2014)

Bottom Line: TGF-β1-stimulated Nox4 expression and ROS formation in endothelial cells.In cells from Nox4-deficient mice, TGF-β1-induced cell proliferation, migration and tube formation were abolished.In vivo, TGF-β1 stimulated growth of blood vessels into sponges implanted subcutaneously, and this angiogenesis was markedly reduced in Nox4 knockout mice.

View Article: PubMed Central - PubMed

Affiliation: Centre for Eye Research Australia, Department of Ophthalmology, University of Melbourne, East Melbourne, Victoria, Australia; O'Brien Institute, Fitzroy, Victoria, Australia.

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Nox4 is required for transforming growth factor-β1 (TGF-β1)-induced endothelial cell capillary formation. Adv-Nox4i abrogated the stimulatory effect of TGF-β1 (10 ng/ml) on capillary formation of (A and B) in HUVECs and (C) HMECs in growth factor reduced Matrigel. Similarly, TGF-β1 (10 ng/ml) induced capillary formation within 8 hrs of application in MHEC derived from wild-type (WT), and this response was abrogated in Nox4 knockout mouse cells (Nox4 KO; D and E). Representative high magnification images of tube formation assay performed on (A) HUVECs and (D) MHEC (scale bar represents 100 μm). Quantitative measures (mean ± SEM from n = 6) of capillary formation are expressed as a percentage of cells infected with Adv-GFP controls or WT cells in the absence of TGF-β1. *P < 0.05 from Adv-GFP or WT without TGF-β1 treatment; †P < 0.05 from Adv-GFP or WT treatment in the presence of TGF-β1.
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fig05: Nox4 is required for transforming growth factor-β1 (TGF-β1)-induced endothelial cell capillary formation. Adv-Nox4i abrogated the stimulatory effect of TGF-β1 (10 ng/ml) on capillary formation of (A and B) in HUVECs and (C) HMECs in growth factor reduced Matrigel. Similarly, TGF-β1 (10 ng/ml) induced capillary formation within 8 hrs of application in MHEC derived from wild-type (WT), and this response was abrogated in Nox4 knockout mouse cells (Nox4 KO; D and E). Representative high magnification images of tube formation assay performed on (A) HUVECs and (D) MHEC (scale bar represents 100 μm). Quantitative measures (mean ± SEM from n = 6) of capillary formation are expressed as a percentage of cells infected with Adv-GFP controls or WT cells in the absence of TGF-β1. *P < 0.05 from Adv-GFP or WT without TGF-β1 treatment; †P < 0.05 from Adv-GFP or WT treatment in the presence of TGF-β1.

Mentions: In other studies TGF-β1 has been suggested to promote angiogenesis [32,33]. To explore the functional significance of TGF-β1 in regulation of Nox4, we therefore examined the effect of TGF-β1 on angiogenic responses of endothelial cells in vitro and then in vivo. Formation of capillary-like structures was assessed by plating HUVEC and HMECs on solidified growth factor-reduced Matrigel in the absence of serum. Within 8 hrs, TGF-β1-treated cells formed capillary-like structures more efficiently than untreated cells as shown in representative pictures of HUVEC in Figure5A. Transforming growth factor β1-enhanced capillary-like structure formation was indeed prevented by Adv-Nox4i treatment in HUVEC and HMECs (Fig.5B and C). To confirm that Nox4 is required for the TGF-β1-induced angiogenesis, we also studied MHECs. Similar as in human endothelial cells, TGF-β1 enhanced capillary-like structures in WT mouse-derived MHEC suspended in Matrigel, and this response was abolished in Nox4 KO mouse-derived MHEC (Fig.5D and E). These findings suggest that TGF-β1-induced Nox4 plays a role in the formation of endothelial capillary-like structures by endothelial cells, at least in vitro.


Transforming growth factor-β1 requires NADPH oxidase 4 for angiogenesis in vitro and in vivo.

Peshavariya HM, Chan EC, Liu GS, Jiang F, Dusting GJ - J. Cell. Mol. Med. (2014)

Nox4 is required for transforming growth factor-β1 (TGF-β1)-induced endothelial cell capillary formation. Adv-Nox4i abrogated the stimulatory effect of TGF-β1 (10 ng/ml) on capillary formation of (A and B) in HUVECs and (C) HMECs in growth factor reduced Matrigel. Similarly, TGF-β1 (10 ng/ml) induced capillary formation within 8 hrs of application in MHEC derived from wild-type (WT), and this response was abrogated in Nox4 knockout mouse cells (Nox4 KO; D and E). Representative high magnification images of tube formation assay performed on (A) HUVECs and (D) MHEC (scale bar represents 100 μm). Quantitative measures (mean ± SEM from n = 6) of capillary formation are expressed as a percentage of cells infected with Adv-GFP controls or WT cells in the absence of TGF-β1. *P < 0.05 from Adv-GFP or WT without TGF-β1 treatment; †P < 0.05 from Adv-GFP or WT treatment in the presence of TGF-β1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4508156&req=5

fig05: Nox4 is required for transforming growth factor-β1 (TGF-β1)-induced endothelial cell capillary formation. Adv-Nox4i abrogated the stimulatory effect of TGF-β1 (10 ng/ml) on capillary formation of (A and B) in HUVECs and (C) HMECs in growth factor reduced Matrigel. Similarly, TGF-β1 (10 ng/ml) induced capillary formation within 8 hrs of application in MHEC derived from wild-type (WT), and this response was abrogated in Nox4 knockout mouse cells (Nox4 KO; D and E). Representative high magnification images of tube formation assay performed on (A) HUVECs and (D) MHEC (scale bar represents 100 μm). Quantitative measures (mean ± SEM from n = 6) of capillary formation are expressed as a percentage of cells infected with Adv-GFP controls or WT cells in the absence of TGF-β1. *P < 0.05 from Adv-GFP or WT without TGF-β1 treatment; †P < 0.05 from Adv-GFP or WT treatment in the presence of TGF-β1.
Mentions: In other studies TGF-β1 has been suggested to promote angiogenesis [32,33]. To explore the functional significance of TGF-β1 in regulation of Nox4, we therefore examined the effect of TGF-β1 on angiogenic responses of endothelial cells in vitro and then in vivo. Formation of capillary-like structures was assessed by plating HUVEC and HMECs on solidified growth factor-reduced Matrigel in the absence of serum. Within 8 hrs, TGF-β1-treated cells formed capillary-like structures more efficiently than untreated cells as shown in representative pictures of HUVEC in Figure5A. Transforming growth factor β1-enhanced capillary-like structure formation was indeed prevented by Adv-Nox4i treatment in HUVEC and HMECs (Fig.5B and C). To confirm that Nox4 is required for the TGF-β1-induced angiogenesis, we also studied MHECs. Similar as in human endothelial cells, TGF-β1 enhanced capillary-like structures in WT mouse-derived MHEC suspended in Matrigel, and this response was abolished in Nox4 KO mouse-derived MHEC (Fig.5D and E). These findings suggest that TGF-β1-induced Nox4 plays a role in the formation of endothelial capillary-like structures by endothelial cells, at least in vitro.

Bottom Line: TGF-β1-stimulated Nox4 expression and ROS formation in endothelial cells.In cells from Nox4-deficient mice, TGF-β1-induced cell proliferation, migration and tube formation were abolished.In vivo, TGF-β1 stimulated growth of blood vessels into sponges implanted subcutaneously, and this angiogenesis was markedly reduced in Nox4 knockout mice.

View Article: PubMed Central - PubMed

Affiliation: Centre for Eye Research Australia, Department of Ophthalmology, University of Melbourne, East Melbourne, Victoria, Australia; O'Brien Institute, Fitzroy, Victoria, Australia.

Show MeSH
Related in: MedlinePlus