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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 promoted angiogenesis in DLLI muscle, as assessed by immunohistochemical staining.(A) Histological analysis of muscle tissues was performed by immunostaining. The specimens were stained with H&E (hematoxylin-eosin). Hypoxia pretreatment of BM-MSCs accelerated the repair of muscle fibers at day 7. At day 14, the hypoxia pretreatment BM-MSCs group exhibited a de novo rebound in the microvasculature. (B) Immunohistochemical staining with a specific antibody recognizing the endothelial marker CD31 revealed the presence of more mature vessels (muscle), predominantly in the hypoxia pretreatment BM-MSCs group rather than in the control group, and (C) the number of vessels was measured.*, ** indicate P<0.05, 0.01 versus control group, respectively.
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pone.0126715.g005: Hypoxia pretreatment of BM-MSCs promoted angiogenesis in DLLI muscle, as assessed by immunohistochemical staining.(A) Histological analysis of muscle tissues was performed by immunostaining. The specimens were stained with H&E (hematoxylin-eosin). Hypoxia pretreatment of BM-MSCs accelerated the repair of muscle fibers at day 7. At day 14, the hypoxia pretreatment BM-MSCs group exhibited a de novo rebound in the microvasculature. (B) Immunohistochemical staining with a specific antibody recognizing the endothelial marker CD31 revealed the presence of more mature vessels (muscle), predominantly in the hypoxia pretreatment BM-MSCs group rather than in the control group, and (C) the number of vessels was measured.*, ** indicate P<0.05, 0.01 versus control group, respectively.

Mentions: The ischemic muscle tissues were collected for paraffin sections. H&E staining indicated that hypoxia pretreatment of BM-MSCs significantly repaired muscle fibers in ischemic tissue at 7 d and 14 d, compared to the other groups (Fig 5A). Next, immunohistochemical staining demonstrated that the mature microvessel density, counted by immunostaining of endothelial cells marked with anti- CD31 antibodyies, was markedly increased in the hypoxia pretreatment BM-MSCs group at 7 d (Fig 5B and 5C). Hypoxia pretreatment of BM-MSCs increased the repair of ischemic tissues in DLLI.


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 promoted angiogenesis in DLLI muscle, as assessed by immunohistochemical staining.(A) Histological analysis of muscle tissues was performed by immunostaining. The specimens were stained with H&E (hematoxylin-eosin). Hypoxia pretreatment of BM-MSCs accelerated the repair of muscle fibers at day 7. At day 14, the hypoxia pretreatment BM-MSCs group exhibited a de novo rebound in the microvasculature. (B) Immunohistochemical staining with a specific antibody recognizing the endothelial marker CD31 revealed the presence of more mature vessels (muscle), predominantly in the hypoxia pretreatment BM-MSCs group rather than in the control group, and (C) the number of vessels was measured.*, ** indicate P<0.05, 0.01 versus control group, respectively.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0126715.g005: Hypoxia pretreatment of BM-MSCs promoted angiogenesis in DLLI muscle, as assessed by immunohistochemical staining.(A) Histological analysis of muscle tissues was performed by immunostaining. The specimens were stained with H&E (hematoxylin-eosin). Hypoxia pretreatment of BM-MSCs accelerated the repair of muscle fibers at day 7. At day 14, the hypoxia pretreatment BM-MSCs group exhibited a de novo rebound in the microvasculature. (B) Immunohistochemical staining with a specific antibody recognizing the endothelial marker CD31 revealed the presence of more mature vessels (muscle), predominantly in the hypoxia pretreatment BM-MSCs group rather than in the control group, and (C) the number of vessels was measured.*, ** indicate P<0.05, 0.01 versus control group, respectively.
Mentions: The ischemic muscle tissues were collected for paraffin sections. H&E staining indicated that hypoxia pretreatment of BM-MSCs significantly repaired muscle fibers in ischemic tissue at 7 d and 14 d, compared to the other groups (Fig 5A). Next, immunohistochemical staining demonstrated that the mature microvessel density, counted by immunostaining of endothelial cells marked with anti- CD31 antibodyies, was markedly increased in the hypoxia pretreatment BM-MSCs group at 7 d (Fig 5B and 5C). Hypoxia pretreatment of BM-MSCs increased the repair of ischemic tissues in DLLI.

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