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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

Time course of perfusion recovery after induced-ischemia treated by Ad/hTK- transduced EPCs.Time course of perfusion recovery after induced-ischemia evaluated by laser doppler flowmetry. Abdominal area and ventral parts of limbs and tail are shown in these digital color-coded images, red indicates regions with maximum perfusion; medium perfusion values are shown in yellow; lowest perfusion values are represented as blue. Images a, d, g and J show the 1st day after rats were injected with cells; images b, e, h and k show on day 14; images c, f, i and l show on day 21. Perfusion recovery during following weeks (days 14 and 21) was accelerated in rat injected with Ad/hTK-transduced EPCs (k, l) compared with Ad/GFP-transduced EPCs (h and i), EPCs (e, f) and 0.9% saline (b, c) (left hindlimb induced- ischemia).
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pone-0073035-g013: Time course of perfusion recovery after induced-ischemia treated by Ad/hTK- transduced EPCs.Time course of perfusion recovery after induced-ischemia evaluated by laser doppler flowmetry. Abdominal area and ventral parts of limbs and tail are shown in these digital color-coded images, red indicates regions with maximum perfusion; medium perfusion values are shown in yellow; lowest perfusion values are represented as blue. Images a, d, g and J show the 1st day after rats were injected with cells; images b, e, h and k show on day 14; images c, f, i and l show on day 21. Perfusion recovery during following weeks (days 14 and 21) was accelerated in rat injected with Ad/hTK-transduced EPCs (k, l) compared with Ad/GFP-transduced EPCs (h and i), EPCs (e, f) and 0.9% saline (b, c) (left hindlimb induced- ischemia).

Mentions: Systolic blood pressure and heart rate were not affected by the femoral artery removal or adenovirus injection (data not shown). The induction of ischemia was followed by a dramatic drop in blood flow. In rats transplanted with EPCs and Ad/GFP-transduced EPCs, this effect was followed by a gradual recovery except for the most distal part of the ischemic hindlimb. Gene therapy with EPCs accelerated the hemodynamic recovery of the whole limb (P<0.01). As shown in Fig. 12 and Fig. 13, the average of plantar blood flow ratio in saline was lower than the flow ratio in EPCs or Ad/hTK-transduced EPCs (0.30±0.07 versus 0.42±0.08, P<0.05 or 0.61±0.08, P<0.01 at day 7; 0.46±0.08 versus 0.61±0.13, P<0.05 or 0.87±0.11, P<0.01 at day 14; 0.50±0.07 versus 0.62±0.05, P<0.01 or 0.91±0.11, P<0.01 at day 21). The average values of plantar blood flow ratio in EPCs were also lower than the average values in Ad/hTK-transduced EPCs, but they presented no significant difference compared with the average values in the Ad/GFP-transduced EPCs. The blood perfusion of the rats injected with Ad/hTK-transduced EPCs exhibited highly accelerated hemodynamic recovery of the whole limb (P<0.01). After 21 days, the blood perfusion of the whole limb in the rats injected with Ad/hTK-transduced EPCs was 54% higher than that in rats injected with saline and was 25% higher than that in rats injected with non-transduced EPCs.


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)

Time course of perfusion recovery after induced-ischemia treated by Ad/hTK- transduced EPCs.Time course of perfusion recovery after induced-ischemia evaluated by laser doppler flowmetry. Abdominal area and ventral parts of limbs and tail are shown in these digital color-coded images, red indicates regions with maximum perfusion; medium perfusion values are shown in yellow; lowest perfusion values are represented as blue. Images a, d, g and J show the 1st day after rats were injected with cells; images b, e, h and k show on day 14; images c, f, i and l show on day 21. Perfusion recovery during following weeks (days 14 and 21) was accelerated in rat injected with Ad/hTK-transduced EPCs (k, l) compared with Ad/GFP-transduced EPCs (h and i), EPCs (e, f) and 0.9% saline (b, c) (left hindlimb induced- ischemia).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3760867&req=5

pone-0073035-g013: Time course of perfusion recovery after induced-ischemia treated by Ad/hTK- transduced EPCs.Time course of perfusion recovery after induced-ischemia evaluated by laser doppler flowmetry. Abdominal area and ventral parts of limbs and tail are shown in these digital color-coded images, red indicates regions with maximum perfusion; medium perfusion values are shown in yellow; lowest perfusion values are represented as blue. Images a, d, g and J show the 1st day after rats were injected with cells; images b, e, h and k show on day 14; images c, f, i and l show on day 21. Perfusion recovery during following weeks (days 14 and 21) was accelerated in rat injected with Ad/hTK-transduced EPCs (k, l) compared with Ad/GFP-transduced EPCs (h and i), EPCs (e, f) and 0.9% saline (b, c) (left hindlimb induced- ischemia).
Mentions: Systolic blood pressure and heart rate were not affected by the femoral artery removal or adenovirus injection (data not shown). The induction of ischemia was followed by a dramatic drop in blood flow. In rats transplanted with EPCs and Ad/GFP-transduced EPCs, this effect was followed by a gradual recovery except for the most distal part of the ischemic hindlimb. Gene therapy with EPCs accelerated the hemodynamic recovery of the whole limb (P<0.01). As shown in Fig. 12 and Fig. 13, the average of plantar blood flow ratio in saline was lower than the flow ratio in EPCs or Ad/hTK-transduced EPCs (0.30±0.07 versus 0.42±0.08, P<0.05 or 0.61±0.08, P<0.01 at day 7; 0.46±0.08 versus 0.61±0.13, P<0.05 or 0.87±0.11, P<0.01 at day 14; 0.50±0.07 versus 0.62±0.05, P<0.01 or 0.91±0.11, P<0.01 at day 21). The average values of plantar blood flow ratio in EPCs were also lower than the average values in Ad/hTK-transduced EPCs, but they presented no significant difference compared with the average values in the Ad/GFP-transduced EPCs. The blood perfusion of the rats injected with Ad/hTK-transduced EPCs exhibited highly accelerated hemodynamic recovery of the whole limb (P<0.01). After 21 days, the blood perfusion of the whole limb in the rats injected with Ad/hTK-transduced EPCs was 54% higher than that in rats injected with saline and was 25% higher than that in rats injected with non-transduced EPCs.

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