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Kallikrein gene-modified EPCs induce angiogenesis in rats with ischemic hindlimb and correlate with integrin αvβ3 expression.

Fu SS, Li FJ, Wang YY, You AB, Qie YL, Meng X, Li JR, Li BC, Zhang Y, Da Li Q - PLoS ONE (2013)

Bottom Line: Expressions of integrin αvβ3 and endothelial nitric oxide synthase (eNOS) were detected on the surface of EPCs.The levels of integrin αvβ3 expression were reduced by PI3K and eNOS blockade, and the inhibitor of integrin αvβ3 abrogated the migration and adhesion of hTK-transduced EPCs (P<0.05). hTK gene delivery in vivo improves the natural angiogenic response to ischemia.The ability of hTK gene-transduced EPCs can be enhanced in vitro, in which integrin αvβ3 plays a role in the process.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Qilu Hospital, Shandong University, Jinan, Shandong Province, China.

ABSTRACT

Background: Human tissue kallikrein (hTK) plays an essential role in the physiological and pathological mechanisms of blood vessels. This study aimed to determine whether angiogenesis induced by endothelial progenitor cells (EPCs) transduced with the adenovirus-mediated hTK gene could improve blood flow in rat hindlimb ischemia in vivo and to establish a promising mechanism in vitro.

Methods: EPCs transduced with adenovirus encoding hTK-162 (i.e., Ad/hTK-transduced EPCs or Ad/GFP-transduced EPCs) were administered to Wister rats with hindlimb ischemia through therapeutic neovascularization. Muscular capillary density (MCD), blood flow (BF), and the number of myofibers were measured at days 7, 14, and 21 after treatment. Expressions of integrin αvβ3 and endothelial nitric oxide synthase (eNOS) were detected on the surface of EPCs.

Results: MCD, BF, and the number of myofibers in rats with Ad/hTK-transduced EPCs remarkably increased at day 21 after treatment compared with rats with Ad/GFP-transduced EPCs or the control group (P<0.01). Expressions of integrin αvβ3 and eNOS protein on the surface of EPCs also increased in rats with Ad/hTK-transduced EPCs. The levels of integrin αvβ3 expression were reduced by PI3K and eNOS blockade, and the inhibitor of integrin αvβ3 abrogated the migration and adhesion of hTK-transduced EPCs (P<0.05).

Conclusion: hTK gene delivery in vivo improves the natural angiogenic response to ischemia. The ability of hTK gene-transduced EPCs can be enhanced in vitro, in which integrin αvβ3 plays a role in the process.

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Related in: MedlinePlus

Collateral capillaries identified by staining with antibodies for smooth muscle actin.Collateral capillaries were identified by staining with antibodies for smooth muscle actin. The ratio of the average number of collateral capillaries in the ischemic thigh to the average number in the non-ischemic thigh was determined for each rat and used as a single data point.*P<0.01 versus control; +P<0.01 versus EPCs.
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pone-0073035-g015: Collateral capillaries identified by staining with antibodies for smooth muscle actin.Collateral capillaries were identified by staining with antibodies for smooth muscle actin. The ratio of the average number of collateral capillaries in the ischemic thigh to the average number in the non-ischemic thigh was determined for each rat and used as a single data point.*P<0.01 versus control; +P<0.01 versus EPCs.

Mentions: The ischemic/non-ischemic collateral capillary ratio was 36% lower in EPCs and 48% lower in saline than that in Ad/hTK-transduced EPCs (Fig. 14). The ratio of collateral myofibro number of ischemic versus non-ischemic was 37% lower than that in EPCs and 54% lower in saline compared with that in Ad/hTK-transduced EPCs (Fig. 15). Capillary densities were significantly greater in Ad/hTK-transduced EPCs and non-transduced EPCs than in saline (P<0.01). The immunohistochemical identification of vascular ECs using antibodies against CD31+ and myofibers through antibodies’ anti-alpha smooth muscle actins is shown in Fig. 16.


Kallikrein gene-modified EPCs induce angiogenesis in rats with ischemic hindlimb and correlate with integrin αvβ3 expression.

Fu SS, Li FJ, Wang YY, You AB, Qie YL, Meng X, Li JR, Li BC, Zhang Y, Da Li Q - PLoS ONE (2013)

Collateral capillaries identified by staining with antibodies for smooth muscle actin.Collateral capillaries were identified by staining with antibodies for smooth muscle actin. The ratio of the average number of collateral capillaries in the ischemic thigh to the average number in the non-ischemic thigh was determined for each rat and used as a single data point.*P<0.01 versus control; +P<0.01 versus EPCs.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3760867&req=5

pone-0073035-g015: Collateral capillaries identified by staining with antibodies for smooth muscle actin.Collateral capillaries were identified by staining with antibodies for smooth muscle actin. The ratio of the average number of collateral capillaries in the ischemic thigh to the average number in the non-ischemic thigh was determined for each rat and used as a single data point.*P<0.01 versus control; +P<0.01 versus EPCs.
Mentions: The ischemic/non-ischemic collateral capillary ratio was 36% lower in EPCs and 48% lower in saline than that in Ad/hTK-transduced EPCs (Fig. 14). The ratio of collateral myofibro number of ischemic versus non-ischemic was 37% lower than that in EPCs and 54% lower in saline compared with that in Ad/hTK-transduced EPCs (Fig. 15). Capillary densities were significantly greater in Ad/hTK-transduced EPCs and non-transduced EPCs than in saline (P<0.01). The immunohistochemical identification of vascular ECs using antibodies against CD31+ and myofibers through antibodies’ anti-alpha smooth muscle actins is shown in Fig. 16.

Bottom Line: Expressions of integrin αvβ3 and endothelial nitric oxide synthase (eNOS) were detected on the surface of EPCs.The levels of integrin αvβ3 expression were reduced by PI3K and eNOS blockade, and the inhibitor of integrin αvβ3 abrogated the migration and adhesion of hTK-transduced EPCs (P<0.05). hTK gene delivery in vivo improves the natural angiogenic response to ischemia.The ability of hTK gene-transduced EPCs can be enhanced in vitro, in which integrin αvβ3 plays a role in the process.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Qilu Hospital, Shandong University, Jinan, Shandong Province, China.

ABSTRACT

Background: Human tissue kallikrein (hTK) plays an essential role in the physiological and pathological mechanisms of blood vessels. This study aimed to determine whether angiogenesis induced by endothelial progenitor cells (EPCs) transduced with the adenovirus-mediated hTK gene could improve blood flow in rat hindlimb ischemia in vivo and to establish a promising mechanism in vitro.

Methods: EPCs transduced with adenovirus encoding hTK-162 (i.e., Ad/hTK-transduced EPCs or Ad/GFP-transduced EPCs) were administered to Wister rats with hindlimb ischemia through therapeutic neovascularization. Muscular capillary density (MCD), blood flow (BF), and the number of myofibers were measured at days 7, 14, and 21 after treatment. Expressions of integrin αvβ3 and endothelial nitric oxide synthase (eNOS) were detected on the surface of EPCs.

Results: MCD, BF, and the number of myofibers in rats with Ad/hTK-transduced EPCs remarkably increased at day 21 after treatment compared with rats with Ad/GFP-transduced EPCs or the control group (P<0.01). Expressions of integrin αvβ3 and eNOS protein on the surface of EPCs also increased in rats with Ad/hTK-transduced EPCs. The levels of integrin αvβ3 expression were reduced by PI3K and eNOS blockade, and the inhibitor of integrin αvβ3 abrogated the migration and adhesion of hTK-transduced EPCs (P<0.05).

Conclusion: hTK gene delivery in vivo improves the natural angiogenic response to ischemia. The ability of hTK gene-transduced EPCs can be enhanced in vitro, in which integrin αvβ3 plays a role in the process.

Show MeSH
Related in: MedlinePlus