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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

EPCs migration assays.A: Representative macroscopic photographs of Ad/hTK-transduced EPCs, Ad/GFP-transduced EPCs after 24 hours in response to growth factors respectively. Left panel shows Ad/hTK-transduced EPCs; right panel, Ad/GFP-transduced EPCs (×100 magnification); B: Quantitative analysis of EPC migration (**P<0.01 versus Ad/GFP transduced-EPCs or EPCs = .
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pone-0073035-g005: EPCs migration assays.A: Representative macroscopic photographs of Ad/hTK-transduced EPCs, Ad/GFP-transduced EPCs after 24 hours in response to growth factors respectively. Left panel shows Ad/hTK-transduced EPCs; right panel, Ad/GFP-transduced EPCs (×100 magnification); B: Quantitative analysis of EPC migration (**P<0.01 versus Ad/GFP transduced-EPCs or EPCs = .

Mentions: The proliferation activity of Ad/hTK-transduced EPCs exceeded that of Ad/GFP-transduced EPCs (0.60±0.02 versus 0.50±0.02, corrected absorbance at 490 nm, P<0.05) and non-transduced EPCs (0.51±0.03, P<0.05) (Fig. 4), which was detected by CCK-8 kits. After 24 h of hTK transduction a 29% increase was observed in the migration activity in Ad/hTK-transduced EPCs than that in non-transduced EPCs (98.60±7.83 versus 76.40±6.20, P<0.01) (Fig. 5). The adhesion activity of Ad/hTK-transduced EPCs in vitro was significantly different from that of Ad/GFP-transduced EPCs or non-transduced EPCs (204.20±10.46 versus 186.40±16.22, P<0.05, or 187.30±16.65, P<0.05) (Fig. 6).


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)

EPCs migration assays.A: Representative macroscopic photographs of Ad/hTK-transduced EPCs, Ad/GFP-transduced EPCs after 24 hours in response to growth factors respectively. Left panel shows Ad/hTK-transduced EPCs; right panel, Ad/GFP-transduced EPCs (×100 magnification); B: Quantitative analysis of EPC migration (**P<0.01 versus Ad/GFP transduced-EPCs or EPCs = .
© Copyright Policy
Related In: Results  -  Collection

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

pone-0073035-g005: EPCs migration assays.A: Representative macroscopic photographs of Ad/hTK-transduced EPCs, Ad/GFP-transduced EPCs after 24 hours in response to growth factors respectively. Left panel shows Ad/hTK-transduced EPCs; right panel, Ad/GFP-transduced EPCs (×100 magnification); B: Quantitative analysis of EPC migration (**P<0.01 versus Ad/GFP transduced-EPCs or EPCs = .
Mentions: The proliferation activity of Ad/hTK-transduced EPCs exceeded that of Ad/GFP-transduced EPCs (0.60±0.02 versus 0.50±0.02, corrected absorbance at 490 nm, P<0.05) and non-transduced EPCs (0.51±0.03, P<0.05) (Fig. 4), which was detected by CCK-8 kits. After 24 h of hTK transduction a 29% increase was observed in the migration activity in Ad/hTK-transduced EPCs than that in non-transduced EPCs (98.60±7.83 versus 76.40±6.20, P<0.01) (Fig. 5). The adhesion activity of Ad/hTK-transduced EPCs in vitro was significantly different from that of Ad/GFP-transduced EPCs or non-transduced EPCs (204.20±10.46 versus 186.40±16.22, P<0.05, or 187.30±16.65, P<0.05) (Fig. 6).

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