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Propionyl-L-Carnitine Enhances Wound Healing and Counteracts Microvascular Endothelial Cell Dysfunction.

Scioli MG, Lo Giudice P, Bielli A, Tarallo V, De Rosa A, De Falco S, Orlandi A - PLoS ONE (2015)

Bottom Line: A daily oral PLC treatment improved skin flap viability and associated with reactive oxygen species (ROS) reduction, inducible nitric oxide synthase (iNOS) and NO up-regulation, accelerated wound healing and increased capillary density, likely favoring dermal angiogenesis by up-regulation for iNOS, vascular endothelial growth factor (VEGF), placental growth factor (PlGF) and reduction of NADPH-oxidase 4 (Nox4) expression.Interestingly, inhibition of β-oxidation counteracted the beneficial effects of PLC on oxidative stress and endothelial dysfunction.The beneficial effects of PLC likely derived from improvement of mitochondrial β-oxidation and reduction of Nox4-mediated oxidative stress and endothelial dysfunction.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedicine and Prevention, Anatomic Pathology, University of Tor Vergata, Rome, Italy.

ABSTRACT

Background: Impaired wound healing represents a high cost for health care systems. Endothelial dysfunction characterizes dermal microangiopathy and contributes to delayed wound healing and chronic ulcers. Endothelial dysfunction impairs cutaneous microvascular blood flow by inducing an imbalance between vasorelaxation and vasoconstriction as a consequence of reduced nitric oxide (NO) production and the increase of oxidative stress and inflammation. Propionyl-L-carnitine (PLC) is a natural derivative of carnitine that has been reported to ameliorate post-ischemic blood flow recovery.

Methods and results: We investigated the effects of PLC in rat skin flap and cutaneous wound healing. A daily oral PLC treatment improved skin flap viability and associated with reactive oxygen species (ROS) reduction, inducible nitric oxide synthase (iNOS) and NO up-regulation, accelerated wound healing and increased capillary density, likely favoring dermal angiogenesis by up-regulation for iNOS, vascular endothelial growth factor (VEGF), placental growth factor (PlGF) and reduction of NADPH-oxidase 4 (Nox4) expression. In serum-deprived human dermal microvascular endothelial cell cultures, PLC ameliorated endothelial dysfunction by increasing iNOS, PlGF, VEGF receptors 1 and 2 expression and NO level. In addition, PLC counteracted serum deprivation-induced impairment of mitochondrial β-oxidation, Nox4 and cellular adhesion molecule (CAM) expression, ROS generation and leukocyte adhesion. Moreover, dermal microvascular endothelial cell dysfunction was prevented by Nox4 inhibition. Interestingly, inhibition of β-oxidation counteracted the beneficial effects of PLC on oxidative stress and endothelial dysfunction.

Conclusion: PLC treatment improved rat skin flap viability, accelerated wound healing and dermal angiogenesis. The beneficial effects of PLC likely derived from improvement of mitochondrial β-oxidation and reduction of Nox4-mediated oxidative stress and endothelial dysfunction. Antioxidant therapy and pharmacological targeting of endothelial dysfunction may represent a promising tool for the treatment of delayed wound healing or chronic ulcers.

No MeSH data available.


Related in: MedlinePlus

Endothelial dysfunction in serum-deprived HMVECs is mediated by NADPH oxidase 4-dependent oxidative stress.(A) Representative blot for Nox4 protein level in HMVECs transfected with non-targeting siRNA (Ctr siRNA) and siNox4. (B) FAD level (β-oxidation impairment) measured as optical density (OD) assay in basal condition (5% FBS) or serum-deprived HMVECs treated with PBS (vehicle, 12h), plumbagin (10μM in PBS, 12h), siNox4 or non-targeting siRNA (Ctr siRNA). (C) ROS level detection by dichlorodihydrofluorescein fluorescence intensity (DCF F.I). (D,E) Real-time PCR of ICAM-1 and VCAM-1 mRNA in basal, vehicle, plumbagin, siNox4 or non-targeting siRNA treated HMVECs. (F) Leukocyte adhesion assay in basal, vehicle, plumbagin, siNox4 or non-targeting siRNA treated HMVECs. t-Student: * and ** indicate p< 0.01 and p< 0.001. Values are expressed as mean ± SEM of three separate experiments. Abbreviations: OD, optical density; HPF, high power field.
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pone.0140697.g007: Endothelial dysfunction in serum-deprived HMVECs is mediated by NADPH oxidase 4-dependent oxidative stress.(A) Representative blot for Nox4 protein level in HMVECs transfected with non-targeting siRNA (Ctr siRNA) and siNox4. (B) FAD level (β-oxidation impairment) measured as optical density (OD) assay in basal condition (5% FBS) or serum-deprived HMVECs treated with PBS (vehicle, 12h), plumbagin (10μM in PBS, 12h), siNox4 or non-targeting siRNA (Ctr siRNA). (C) ROS level detection by dichlorodihydrofluorescein fluorescence intensity (DCF F.I). (D,E) Real-time PCR of ICAM-1 and VCAM-1 mRNA in basal, vehicle, plumbagin, siNox4 or non-targeting siRNA treated HMVECs. (F) Leukocyte adhesion assay in basal, vehicle, plumbagin, siNox4 or non-targeting siRNA treated HMVECs. t-Student: * and ** indicate p< 0.01 and p< 0.001. Values are expressed as mean ± SEM of three separate experiments. Abbreviations: OD, optical density; HPF, high power field.

Mentions: To verify if Nox4 activity was responsible for oxidative stress in serum-deprived HMVECs, we used the specific Nox4 inhibitor plumbagin [38] and siRNA for Nox4 (siNox4). Specific knockdown of Nox4 was evaluated by blot analysis (Fig 7A). Plumbagin or siNox4 counteracted serum deprivation-induced oxidative stress, as FAD and ROS accumulation (Fig 7B and 7C, p< 0.0001). Plumbagin and siNox4 also prevented serum deprivation-induced increase of ICAM-1 and VCAM-1 expression (Fig 7D and 7E, p< 0.001 and p< 0.01, respectively), as well as leukocyte adhesion (Fig 7F, p< 0.001). Non-targeting siRNA (Ctr siRNA) had no effect. These findings strongly suggest that oxidative stress-induced endothelial dysfunction is mediated by Nox4 activity.


Propionyl-L-Carnitine Enhances Wound Healing and Counteracts Microvascular Endothelial Cell Dysfunction.

Scioli MG, Lo Giudice P, Bielli A, Tarallo V, De Rosa A, De Falco S, Orlandi A - PLoS ONE (2015)

Endothelial dysfunction in serum-deprived HMVECs is mediated by NADPH oxidase 4-dependent oxidative stress.(A) Representative blot for Nox4 protein level in HMVECs transfected with non-targeting siRNA (Ctr siRNA) and siNox4. (B) FAD level (β-oxidation impairment) measured as optical density (OD) assay in basal condition (5% FBS) or serum-deprived HMVECs treated with PBS (vehicle, 12h), plumbagin (10μM in PBS, 12h), siNox4 or non-targeting siRNA (Ctr siRNA). (C) ROS level detection by dichlorodihydrofluorescein fluorescence intensity (DCF F.I). (D,E) Real-time PCR of ICAM-1 and VCAM-1 mRNA in basal, vehicle, plumbagin, siNox4 or non-targeting siRNA treated HMVECs. (F) Leukocyte adhesion assay in basal, vehicle, plumbagin, siNox4 or non-targeting siRNA treated HMVECs. t-Student: * and ** indicate p< 0.01 and p< 0.001. Values are expressed as mean ± SEM of three separate experiments. Abbreviations: OD, optical density; HPF, high power field.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608702&req=5

pone.0140697.g007: Endothelial dysfunction in serum-deprived HMVECs is mediated by NADPH oxidase 4-dependent oxidative stress.(A) Representative blot for Nox4 protein level in HMVECs transfected with non-targeting siRNA (Ctr siRNA) and siNox4. (B) FAD level (β-oxidation impairment) measured as optical density (OD) assay in basal condition (5% FBS) or serum-deprived HMVECs treated with PBS (vehicle, 12h), plumbagin (10μM in PBS, 12h), siNox4 or non-targeting siRNA (Ctr siRNA). (C) ROS level detection by dichlorodihydrofluorescein fluorescence intensity (DCF F.I). (D,E) Real-time PCR of ICAM-1 and VCAM-1 mRNA in basal, vehicle, plumbagin, siNox4 or non-targeting siRNA treated HMVECs. (F) Leukocyte adhesion assay in basal, vehicle, plumbagin, siNox4 or non-targeting siRNA treated HMVECs. t-Student: * and ** indicate p< 0.01 and p< 0.001. Values are expressed as mean ± SEM of three separate experiments. Abbreviations: OD, optical density; HPF, high power field.
Mentions: To verify if Nox4 activity was responsible for oxidative stress in serum-deprived HMVECs, we used the specific Nox4 inhibitor plumbagin [38] and siRNA for Nox4 (siNox4). Specific knockdown of Nox4 was evaluated by blot analysis (Fig 7A). Plumbagin or siNox4 counteracted serum deprivation-induced oxidative stress, as FAD and ROS accumulation (Fig 7B and 7C, p< 0.0001). Plumbagin and siNox4 also prevented serum deprivation-induced increase of ICAM-1 and VCAM-1 expression (Fig 7D and 7E, p< 0.001 and p< 0.01, respectively), as well as leukocyte adhesion (Fig 7F, p< 0.001). Non-targeting siRNA (Ctr siRNA) had no effect. These findings strongly suggest that oxidative stress-induced endothelial dysfunction is mediated by Nox4 activity.

Bottom Line: A daily oral PLC treatment improved skin flap viability and associated with reactive oxygen species (ROS) reduction, inducible nitric oxide synthase (iNOS) and NO up-regulation, accelerated wound healing and increased capillary density, likely favoring dermal angiogenesis by up-regulation for iNOS, vascular endothelial growth factor (VEGF), placental growth factor (PlGF) and reduction of NADPH-oxidase 4 (Nox4) expression.Interestingly, inhibition of β-oxidation counteracted the beneficial effects of PLC on oxidative stress and endothelial dysfunction.The beneficial effects of PLC likely derived from improvement of mitochondrial β-oxidation and reduction of Nox4-mediated oxidative stress and endothelial dysfunction.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedicine and Prevention, Anatomic Pathology, University of Tor Vergata, Rome, Italy.

ABSTRACT

Background: Impaired wound healing represents a high cost for health care systems. Endothelial dysfunction characterizes dermal microangiopathy and contributes to delayed wound healing and chronic ulcers. Endothelial dysfunction impairs cutaneous microvascular blood flow by inducing an imbalance between vasorelaxation and vasoconstriction as a consequence of reduced nitric oxide (NO) production and the increase of oxidative stress and inflammation. Propionyl-L-carnitine (PLC) is a natural derivative of carnitine that has been reported to ameliorate post-ischemic blood flow recovery.

Methods and results: We investigated the effects of PLC in rat skin flap and cutaneous wound healing. A daily oral PLC treatment improved skin flap viability and associated with reactive oxygen species (ROS) reduction, inducible nitric oxide synthase (iNOS) and NO up-regulation, accelerated wound healing and increased capillary density, likely favoring dermal angiogenesis by up-regulation for iNOS, vascular endothelial growth factor (VEGF), placental growth factor (PlGF) and reduction of NADPH-oxidase 4 (Nox4) expression. In serum-deprived human dermal microvascular endothelial cell cultures, PLC ameliorated endothelial dysfunction by increasing iNOS, PlGF, VEGF receptors 1 and 2 expression and NO level. In addition, PLC counteracted serum deprivation-induced impairment of mitochondrial β-oxidation, Nox4 and cellular adhesion molecule (CAM) expression, ROS generation and leukocyte adhesion. Moreover, dermal microvascular endothelial cell dysfunction was prevented by Nox4 inhibition. Interestingly, inhibition of β-oxidation counteracted the beneficial effects of PLC on oxidative stress and endothelial dysfunction.

Conclusion: PLC treatment improved rat skin flap viability, accelerated wound healing and dermal angiogenesis. The beneficial effects of PLC likely derived from improvement of mitochondrial β-oxidation and reduction of Nox4-mediated oxidative stress and endothelial dysfunction. Antioxidant therapy and pharmacological targeting of endothelial dysfunction may represent a promising tool for the treatment of delayed wound healing or chronic ulcers.

No MeSH data available.


Related in: MedlinePlus