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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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Rat bone marrow EPCs isolation.The morphological characteristics of mononuclear cells from bone marrow (MCBM) were cultured in differential medium with time: (A) after cultured days 3; (B): days 7, the MCBMs differentiated into EPCs; (C): days 10; (D).capillary-like structures formed on culture bottle at days 21 (×100 magnification).
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pone-0073035-g001: Rat bone marrow EPCs isolation.The morphological characteristics of mononuclear cells from bone marrow (MCBM) were cultured in differential medium with time: (A) after cultured days 3; (B): days 7, the MCBMs differentiated into EPCs; (C): days 10; (D).capillary-like structures formed on culture bottle at days 21 (×100 magnification).

Mentions: Total MCBMs isolated and cultured for days 1, 3, 7 and 10 resulted in a spindle-shaped, PEC-like morphology (Fig. 1). After one week, most of cultured cells demonstrated a typical spindle-shaped morphology. Such morphological appearance resembled that of endothelial progenitor cells developed from the MCBM cells. Cellular immunostaining demonstrated that most adherent cells presented double positive staining for an uptake of Dil-acLDL and binding of FITC-UEA-1, which indicate the nature of EPCs (Fig. 2A). These cells positively expressed endothelial cell-specific antigens (KDR and CD31), and a small fraction of the adherent cells expressed marker CD34 and CD133 (Fig. 2B and Fig. 2C).


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)

Rat bone marrow EPCs isolation.The morphological characteristics of mononuclear cells from bone marrow (MCBM) were cultured in differential medium with time: (A) after cultured days 3; (B): days 7, the MCBMs differentiated into EPCs; (C): days 10; (D).capillary-like structures formed on culture bottle at days 21 (×100 magnification).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0073035-g001: Rat bone marrow EPCs isolation.The morphological characteristics of mononuclear cells from bone marrow (MCBM) were cultured in differential medium with time: (A) after cultured days 3; (B): days 7, the MCBMs differentiated into EPCs; (C): days 10; (D).capillary-like structures formed on culture bottle at days 21 (×100 magnification).
Mentions: Total MCBMs isolated and cultured for days 1, 3, 7 and 10 resulted in a spindle-shaped, PEC-like morphology (Fig. 1). After one week, most of cultured cells demonstrated a typical spindle-shaped morphology. Such morphological appearance resembled that of endothelial progenitor cells developed from the MCBM cells. Cellular immunostaining demonstrated that most adherent cells presented double positive staining for an uptake of Dil-acLDL and binding of FITC-UEA-1, which indicate the nature of EPCs (Fig. 2A). These cells positively expressed endothelial cell-specific antigens (KDR and CD31), and a small fraction of the adherent cells expressed marker CD34 and CD133 (Fig. 2B and Fig. 2C).

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