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Hypoxia pretreatment of bone marrow mesenchymal stem cells facilitates angiogenesis by improving the function of endothelial cells in diabetic rats with lower ischemia.

Liu J, Hao H, Xia L, Ti D, Huang H, Dong L, Tong C, Hou Q, Zhao Y, Liu H, Fu X, Han W - PLoS ONE (2015)

Bottom Line: Endothelial dysfunction induced by unordered metabolism results in vascular reconstruction challenges in diabetic lower limb ischemia (DLLI).CM-Dil-labeled tracer experiments indicated that the survival of BM-MSCs was significantly improved, with approximately 5% of the injected cells remaining alive at 14 days.The expression levels of VEGF-1α, MMP-9 and VEGF-R were significantly increased, and the expression of pAKT was up-regulated in ischemic muscle.

View Article: PubMed Central - PubMed

Affiliation: Institute of Basic Medicine Science, College of Life Science, Chinese PLA General Hospital, Beijing, China.

ABSTRACT
Endothelial dysfunction induced by unordered metabolism results in vascular reconstruction challenges in diabetic lower limb ischemia (DLLI). Mesenchymal stem cells (MSCs) are multipotent secretory cells that are suitable for clinical DLLI treatment, but their use has been hampered by poor survival after injection. Hypoxia can significantly enhance the capacity of MSCs to secrete angiogenic factors. We investigated transient hypoxia pretreatment of MSCs to facilitate revascularization in DLLI. Rat bone marrow MSCs (BM-MSCs) were cultured at different oxygen concentrations for varying time periods. The results indicated that transient pretreatment (5% O2, 48 h) not only increased the expression of VEGF-1α, ANG, HIF-1α and MMP-9 in BM-MSCs as assessed by real-time RT-PCR, but also increased the expression of Bcl-2 as determined by western blotting. The transplantation of pretreated BM-MSCs into rats with DLLI demonstrated accelerated vascular reconstruction when assayed by angiography and immunohistochemistry. CM-Dil-labeled tracer experiments indicated that the survival of BM-MSCs was significantly improved, with approximately 5% of the injected cells remaining alive at 14 days. The expression levels of VEGF-1α, MMP-9 and VEGF-R were significantly increased, and the expression of pAKT was up-regulated in ischemic muscle. Double immunofluorescence studies confirmed that the pretreated BM-MSCs promoted the proliferation and inhibited the apoptosis of endothelial cells. In vitro, pretreated BM-MSCs increased the migratory and tube forming capacity of endothelial cells (ECs). Hypoxia pretreatment of BM-MSCs significantly improved angiogenesis in response to tissue ischemia by ameliorating endothelial cell dysfunction and is a promising therapeutic treatment for DLLI.

No MeSH data available.


Related in: MedlinePlus

Hypoxia pretreatment of BM-MSCs modulates VEGF/AKT signaling in HUVECs.(A) The expression of VEGF-1α was suppressed with lentiviral-mediated siRNA-VEGF-1α (siVEGF). (B) CM generated from hypoxia pretreated BM-MSCs treated with siVEGF downregulated the expression of pAKT and VEGFR in HUVECs, as determined by western blot. (C) The capacity of HUVECs to form tubes was inhibited by CM from VEGF-1α-silenced BM-MSCs.
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pone.0126715.g003: Hypoxia pretreatment of BM-MSCs modulates VEGF/AKT signaling in HUVECs.(A) The expression of VEGF-1α was suppressed with lentiviral-mediated siRNA-VEGF-1α (siVEGF). (B) CM generated from hypoxia pretreated BM-MSCs treated with siVEGF downregulated the expression of pAKT and VEGFR in HUVECs, as determined by western blot. (C) The capacity of HUVECs to form tubes was inhibited by CM from VEGF-1α-silenced BM-MSCs.

Mentions: The expressions of VEGF/AKT-related proteins in endothelial cells were affected by hypoxia pretreatment of BM-MSCs, as assessed by western blot. The results indicated that the expression of pAKT and VEGF-R in ECs was upregulated with H-CM. However, when the expression of VEGF-1α was silenced in BM-MSCs by lentivirus-mediated siRNA (Fig 3A), the upregulated expression of VEGFR and pAKT was suppressed in HUVECs treated with H-siVEGF-CM. (Fig 3B). Furthermore, the capacity for tube formation by HUVECs was inhibited by H-siVEGF-CM treatment (Fig 3C). The expression of VEGF/AKT-related protein in ECs was activated by BM-MSCs, and the capacity for angiogenesis was improved.


Hypoxia pretreatment of bone marrow mesenchymal stem cells facilitates angiogenesis by improving the function of endothelial cells in diabetic rats with lower ischemia.

Liu J, Hao H, Xia L, Ti D, Huang H, Dong L, Tong C, Hou Q, Zhao Y, Liu H, Fu X, Han W - PLoS ONE (2015)

Hypoxia pretreatment of BM-MSCs modulates VEGF/AKT signaling in HUVECs.(A) The expression of VEGF-1α was suppressed with lentiviral-mediated siRNA-VEGF-1α (siVEGF). (B) CM generated from hypoxia pretreated BM-MSCs treated with siVEGF downregulated the expression of pAKT and VEGFR in HUVECs, as determined by western blot. (C) The capacity of HUVECs to form tubes was inhibited by CM from VEGF-1α-silenced BM-MSCs.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440823&req=5

pone.0126715.g003: Hypoxia pretreatment of BM-MSCs modulates VEGF/AKT signaling in HUVECs.(A) The expression of VEGF-1α was suppressed with lentiviral-mediated siRNA-VEGF-1α (siVEGF). (B) CM generated from hypoxia pretreated BM-MSCs treated with siVEGF downregulated the expression of pAKT and VEGFR in HUVECs, as determined by western blot. (C) The capacity of HUVECs to form tubes was inhibited by CM from VEGF-1α-silenced BM-MSCs.
Mentions: The expressions of VEGF/AKT-related proteins in endothelial cells were affected by hypoxia pretreatment of BM-MSCs, as assessed by western blot. The results indicated that the expression of pAKT and VEGF-R in ECs was upregulated with H-CM. However, when the expression of VEGF-1α was silenced in BM-MSCs by lentivirus-mediated siRNA (Fig 3A), the upregulated expression of VEGFR and pAKT was suppressed in HUVECs treated with H-siVEGF-CM. (Fig 3B). Furthermore, the capacity for tube formation by HUVECs was inhibited by H-siVEGF-CM treatment (Fig 3C). The expression of VEGF/AKT-related protein in ECs was activated by BM-MSCs, and the capacity for angiogenesis was improved.

Bottom Line: Endothelial dysfunction induced by unordered metabolism results in vascular reconstruction challenges in diabetic lower limb ischemia (DLLI).CM-Dil-labeled tracer experiments indicated that the survival of BM-MSCs was significantly improved, with approximately 5% of the injected cells remaining alive at 14 days.The expression levels of VEGF-1α, MMP-9 and VEGF-R were significantly increased, and the expression of pAKT was up-regulated in ischemic muscle.

View Article: PubMed Central - PubMed

Affiliation: Institute of Basic Medicine Science, College of Life Science, Chinese PLA General Hospital, Beijing, China.

ABSTRACT
Endothelial dysfunction induced by unordered metabolism results in vascular reconstruction challenges in diabetic lower limb ischemia (DLLI). Mesenchymal stem cells (MSCs) are multipotent secretory cells that are suitable for clinical DLLI treatment, but their use has been hampered by poor survival after injection. Hypoxia can significantly enhance the capacity of MSCs to secrete angiogenic factors. We investigated transient hypoxia pretreatment of MSCs to facilitate revascularization in DLLI. Rat bone marrow MSCs (BM-MSCs) were cultured at different oxygen concentrations for varying time periods. The results indicated that transient pretreatment (5% O2, 48 h) not only increased the expression of VEGF-1α, ANG, HIF-1α and MMP-9 in BM-MSCs as assessed by real-time RT-PCR, but also increased the expression of Bcl-2 as determined by western blotting. The transplantation of pretreated BM-MSCs into rats with DLLI demonstrated accelerated vascular reconstruction when assayed by angiography and immunohistochemistry. CM-Dil-labeled tracer experiments indicated that the survival of BM-MSCs was significantly improved, with approximately 5% of the injected cells remaining alive at 14 days. The expression levels of VEGF-1α, MMP-9 and VEGF-R were significantly increased, and the expression of pAKT was up-regulated in ischemic muscle. Double immunofluorescence studies confirmed that the pretreated BM-MSCs promoted the proliferation and inhibited the apoptosis of endothelial cells. In vitro, pretreated BM-MSCs increased the migratory and tube forming capacity of endothelial cells (ECs). Hypoxia pretreatment of BM-MSCs significantly improved angiogenesis in response to tissue ischemia by ameliorating endothelial cell dysfunction and is a promising therapeutic treatment for DLLI.

No MeSH data available.


Related in: MedlinePlus