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The redox enzyme p66Shc contributes to diabetes and ischemia-induced delay in cutaneous wound healing.

Fadini GP, Albiero M, Menegazzo L, Boscaro E, Pagnin E, Iori E, Cosma C, Lapolla A, Pengo V, Stendardo M, Agostini C, Pelicci PG, Giorgio M, Avogaro A - Diabetes (2010)

Bottom Line: Genetic deletion of p66Shc prolongs life span and protects against oxidative stress.Migration of p66Shc(-/-) dermal fibroblasts in vitro was significantly faster than WT fibroblasts under both high glucose and hypoxia. p66Shc is involved in the delayed wound-healing process in the setting of diabetes and ischemia.Thus, p66Shc may represent a potential therapeutic target against this disabling diabetes complication.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Experimental Medicine, University of Padova Medical School, Padova, Italy. gianpaolofadini@hotmail.com

ABSTRACT

Objective: The redox enzyme p66Shc produces hydrogen peroxide and triggers proapoptotic signals. Genetic deletion of p66Shc prolongs life span and protects against oxidative stress. In the present study, we evaluated the role of p66Shc in an animal model of diabetic wound healing.

Research design and methods: Skin wounds were created in wild-type (WT) and p66Shc(-/-) control and streptozotocin-induced diabetic mice with or without hind limb ischemia. Wounds were assessed for collagen content, thickness and vascularity of granulation tissue, apoptosis, reepithelialization, and expression of c-myc and beta-catenin. Response to hind limb ischemia was also evaluated.

Results: Diabetes delayed wound healing in WT mice with reduced granulation tissue thickness and vascularity, increased apoptosis, epithelial expression of c-myc, and nuclear localization of beta-catenin. These nonhealing features were worsened by hind limb ischemia. Diabetes induced p66Shc expression and activation; wound healing was significantly faster in p66Shc(-/-) than in WT diabetic mice, with or without hind limb ischemia, at 1 and 3 months of diabetes duration and in both SV129 and C57BL/6 genetic backgrounds. Deletion of p66Shc reversed nonhealing features, with increased collagen content and granulation tissue thickness, and reduced apoptosis and expression of c-myc and beta-catenin. p66Shc deletion improved response to hind limb ischemia in diabetic mice in terms of tissue damage, capillary density, and perfusion. Migration of p66Shc(-/-) dermal fibroblasts in vitro was significantly faster than WT fibroblasts under both high glucose and hypoxia.

Conclusions: p66Shc is involved in the delayed wound-healing process in the setting of diabetes and ischemia. Thus, p66Shc may represent a potential therapeutic target against this disabling diabetes complication.

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Related in: MedlinePlus

Wound healing. Healing of a 4-mm–diameter cutaneous wound was monitored using digital photography in wild-type (WT) and p66Shc-knockout (KO) diabetic (DM) and nondiabetic (ND) mice with or without hind limb ischemia. Representative photos are shown. Wound size is reported as fraction of the initial wound area. On the right, healing time is shown for WT and p66Shc−/− mice with or without diabetes and limb ischemia on the SV129 or the C57BLC57BL/6 background at 4 or 12 weeks of diabetes duration. n ≥ 3 mice for each group. *P < 0.05 in KO vs. WT. (A high-quality digital representation of this figure is available in the online issue.)
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Figure 1: Wound healing. Healing of a 4-mm–diameter cutaneous wound was monitored using digital photography in wild-type (WT) and p66Shc-knockout (KO) diabetic (DM) and nondiabetic (ND) mice with or without hind limb ischemia. Representative photos are shown. Wound size is reported as fraction of the initial wound area. On the right, healing time is shown for WT and p66Shc−/− mice with or without diabetes and limb ischemia on the SV129 or the C57BLC57BL/6 background at 4 or 12 weeks of diabetes duration. n ≥ 3 mice for each group. *P < 0.05 in KO vs. WT. (A high-quality digital representation of this figure is available in the online issue.)

Mentions: Wounds 4 mm in diameter were created on the dorsal surface of the right hind limb in WT and p66Shc−/− SV129 mice, in the presence of either 4-week STZ-induced diabetes, 2-week hind limb ischemia, or both. In separated experiments, wounds and ischemia were induced in WT and p66Shc−/− C57BL/6 mice after 4 and 12 weeks of diabetes and in coeval nondiabetic C57BL/6 mice. Wound area was monitored with a digital camera till complete macroscopic closure. As shown in Fig. 1, with both the SV129 and C57BL/6 backgrounds, in nondiabetic nonischemic animals, wound healing was not different between WT and p66Shc−/− mice. Four-week diabetes delayed wound healing by about 70% in WT SV129 animals and 30% in p66Shc−/− animals, resulting in a significantly faster healing in p66Shc−/− than in WT diabetic nonischemic mice (wound area at day 4: 23.6 ± 2.2 vs. 35.9 ± 5.0%; P = 0.028). Hind limb ischemia delayed wound healing by about 185% in WT and 70% in p66Shc−/− mice, resulting in a faster healing in p66Shc−/− versus WT nondiabetic ischemic mice (P < 0.05 from day 1 to 11). The coexistence of both diabetes and hind limb ischemia further exacerbated the differences between WT and p66Shc−/− mice. Similarly, p66Shc deletion in C57BL/6 mice prevented the delay in wound healing induced by 4-week diabetes and hind limb ischemia. In ischemic and nonischemic C57BL/6 mice 12 weeks after diabetes induction, absence of p66Shc was associated with significantly faster wound healing compared with the WT micce (Fig. 1, right panels). With this diabetes duration and genetic background, induction of ischemia was often followed by hind limb auto-amputation in WT mice (30 and 50% in nondiabetic and diabetic animals, respectively), while no amputation was seen in p66Shc−/− mice (supplementary Fig. II).


The redox enzyme p66Shc contributes to diabetes and ischemia-induced delay in cutaneous wound healing.

Fadini GP, Albiero M, Menegazzo L, Boscaro E, Pagnin E, Iori E, Cosma C, Lapolla A, Pengo V, Stendardo M, Agostini C, Pelicci PG, Giorgio M, Avogaro A - Diabetes (2010)

Wound healing. Healing of a 4-mm–diameter cutaneous wound was monitored using digital photography in wild-type (WT) and p66Shc-knockout (KO) diabetic (DM) and nondiabetic (ND) mice with or without hind limb ischemia. Representative photos are shown. Wound size is reported as fraction of the initial wound area. On the right, healing time is shown for WT and p66Shc−/− mice with or without diabetes and limb ischemia on the SV129 or the C57BLC57BL/6 background at 4 or 12 weeks of diabetes duration. n ≥ 3 mice for each group. *P < 0.05 in KO vs. WT. (A high-quality digital representation of this figure is available in the online issue.)
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Figure 1: Wound healing. Healing of a 4-mm–diameter cutaneous wound was monitored using digital photography in wild-type (WT) and p66Shc-knockout (KO) diabetic (DM) and nondiabetic (ND) mice with or without hind limb ischemia. Representative photos are shown. Wound size is reported as fraction of the initial wound area. On the right, healing time is shown for WT and p66Shc−/− mice with or without diabetes and limb ischemia on the SV129 or the C57BLC57BL/6 background at 4 or 12 weeks of diabetes duration. n ≥ 3 mice for each group. *P < 0.05 in KO vs. WT. (A high-quality digital representation of this figure is available in the online issue.)
Mentions: Wounds 4 mm in diameter were created on the dorsal surface of the right hind limb in WT and p66Shc−/− SV129 mice, in the presence of either 4-week STZ-induced diabetes, 2-week hind limb ischemia, or both. In separated experiments, wounds and ischemia were induced in WT and p66Shc−/− C57BL/6 mice after 4 and 12 weeks of diabetes and in coeval nondiabetic C57BL/6 mice. Wound area was monitored with a digital camera till complete macroscopic closure. As shown in Fig. 1, with both the SV129 and C57BL/6 backgrounds, in nondiabetic nonischemic animals, wound healing was not different between WT and p66Shc−/− mice. Four-week diabetes delayed wound healing by about 70% in WT SV129 animals and 30% in p66Shc−/− animals, resulting in a significantly faster healing in p66Shc−/− than in WT diabetic nonischemic mice (wound area at day 4: 23.6 ± 2.2 vs. 35.9 ± 5.0%; P = 0.028). Hind limb ischemia delayed wound healing by about 185% in WT and 70% in p66Shc−/− mice, resulting in a faster healing in p66Shc−/− versus WT nondiabetic ischemic mice (P < 0.05 from day 1 to 11). The coexistence of both diabetes and hind limb ischemia further exacerbated the differences between WT and p66Shc−/− mice. Similarly, p66Shc deletion in C57BL/6 mice prevented the delay in wound healing induced by 4-week diabetes and hind limb ischemia. In ischemic and nonischemic C57BL/6 mice 12 weeks after diabetes induction, absence of p66Shc was associated with significantly faster wound healing compared with the WT micce (Fig. 1, right panels). With this diabetes duration and genetic background, induction of ischemia was often followed by hind limb auto-amputation in WT mice (30 and 50% in nondiabetic and diabetic animals, respectively), while no amputation was seen in p66Shc−/− mice (supplementary Fig. II).

Bottom Line: Genetic deletion of p66Shc prolongs life span and protects against oxidative stress.Migration of p66Shc(-/-) dermal fibroblasts in vitro was significantly faster than WT fibroblasts under both high glucose and hypoxia. p66Shc is involved in the delayed wound-healing process in the setting of diabetes and ischemia.Thus, p66Shc may represent a potential therapeutic target against this disabling diabetes complication.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Experimental Medicine, University of Padova Medical School, Padova, Italy. gianpaolofadini@hotmail.com

ABSTRACT

Objective: The redox enzyme p66Shc produces hydrogen peroxide and triggers proapoptotic signals. Genetic deletion of p66Shc prolongs life span and protects against oxidative stress. In the present study, we evaluated the role of p66Shc in an animal model of diabetic wound healing.

Research design and methods: Skin wounds were created in wild-type (WT) and p66Shc(-/-) control and streptozotocin-induced diabetic mice with or without hind limb ischemia. Wounds were assessed for collagen content, thickness and vascularity of granulation tissue, apoptosis, reepithelialization, and expression of c-myc and beta-catenin. Response to hind limb ischemia was also evaluated.

Results: Diabetes delayed wound healing in WT mice with reduced granulation tissue thickness and vascularity, increased apoptosis, epithelial expression of c-myc, and nuclear localization of beta-catenin. These nonhealing features were worsened by hind limb ischemia. Diabetes induced p66Shc expression and activation; wound healing was significantly faster in p66Shc(-/-) than in WT diabetic mice, with or without hind limb ischemia, at 1 and 3 months of diabetes duration and in both SV129 and C57BL/6 genetic backgrounds. Deletion of p66Shc reversed nonhealing features, with increased collagen content and granulation tissue thickness, and reduced apoptosis and expression of c-myc and beta-catenin. p66Shc deletion improved response to hind limb ischemia in diabetic mice in terms of tissue damage, capillary density, and perfusion. Migration of p66Shc(-/-) dermal fibroblasts in vitro was significantly faster than WT fibroblasts under both high glucose and hypoxia.

Conclusions: p66Shc is involved in the delayed wound-healing process in the setting of diabetes and ischemia. Thus, p66Shc may represent a potential therapeutic target against this disabling diabetes complication.

Show MeSH
Related in: MedlinePlus