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Compromised Wound Healing in Ischemic Type 2 Diabetic Rats.

Yang P, Pei Q, Yu T, Chang Q, Wang D, Gao M, Zhang X, Liu Y - PLoS ONE (2016)

Bottom Line: A bipedicle flap, with length to width ratio 1.5, was performed on the back of the rat to make the flap area ischemic.Our results demonstrated that a combination of HFD feeding and a low dose of STZ is capable of inducing the rats to develop type 2 diabetes with noticeable insulin resistance, persistent hyperglycemia, moderate degree of insulinemia, as well as high serum cholesterol and high triglyceride levels.Our study suggested that HFD feeding combined with STZ injection could induce type 2 diabetes in rat.

View Article: PubMed Central - PubMed

Affiliation: Department of Burn and Plastic Surgery, Shanghai Jiao Tong University School of Medicine, Ruijin Hospital, Shanghai 200025, China.

ABSTRACT
Ischemia is one of the main epidemic factors and characteristics of diabetic chronic wounds, and exerts a profound effect on wound healing. To explore the mechanism of and the cure for diabetic impaired wound healing, we established a type 2 diabetic rat model. We used an 8 weeks high fat diet (HFD) feeding regimen followed by multiple injections of streptozotocin (STZ) at a dose of 10mg/kg to induce Wister rat to develop type 2 diabetes. Metabolic characteristics were assessed at the 5th week after the STZ injections to confirm the establishment of diabetes mellitus on the rodent model. A bipedicle flap, with length to width ratio 1.5, was performed on the back of the rat to make the flap area ischemic. Closure of excisional wounds on this bipedicle flap and related physiological and pathological changes were studied using histological, immunohistochemical, real time PCR and protein immunoblot approaches. Our results demonstrated that a combination of HFD feeding and a low dose of STZ is capable of inducing the rats to develop type 2 diabetes with noticeable insulin resistance, persistent hyperglycemia, moderate degree of insulinemia, as well as high serum cholesterol and high triglyceride levels. The excision wounds on the ischemic double pedicle flap showed deteriorative healing features comparing with non-ischemic diabetic wounds, including: delayed healing, exorbitant wound inflammatory response, excessive and prolonged ROS production and excessive production of MMPs. Our study suggested that HFD feeding combined with STZ injection could induce type 2 diabetes in rat. Our ischemic diabetic wound model is suitable for the investigation of human diabetic related wound repair; especically for diabetic chronic wounds.

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Ischemia induced exorbitant wound inflammatory response.(A) Representative images of MPO immunochemistry staining for both non-ischemic and ischemic wounds at day 5 after wounding. Scale bars = 200μm. (B) Representative images of immunohischemical staining for CD68, a specific marker of macrophage. Scale bars = 200μm. (C-F) Real time PCR analyze TNF-α, IL-6, IL-1β and IL-10 RNA expression (fold change) of non-ischemic and ischemic wounds at day 5 and 11 after wounding. Data was obtained from 12 wounds (n = 6), and are shown as the mean ± SD. **p<0.01. (G-I) Western blot analyze of wounds TNF-α, IL-6 and IL-10 at day 5 and 11 after wounding. Data was obtained from 12 wounds (n = 6), and are shown as the mean ± SD. *P<0.05, **p<0.01.
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pone.0152068.g005: Ischemia induced exorbitant wound inflammatory response.(A) Representative images of MPO immunochemistry staining for both non-ischemic and ischemic wounds at day 5 after wounding. Scale bars = 200μm. (B) Representative images of immunohischemical staining for CD68, a specific marker of macrophage. Scale bars = 200μm. (C-F) Real time PCR analyze TNF-α, IL-6, IL-1β and IL-10 RNA expression (fold change) of non-ischemic and ischemic wounds at day 5 and 11 after wounding. Data was obtained from 12 wounds (n = 6), and are shown as the mean ± SD. **p<0.01. (G-I) Western blot analyze of wounds TNF-α, IL-6 and IL-10 at day 5 and 11 after wounding. Data was obtained from 12 wounds (n = 6), and are shown as the mean ± SD. *P<0.05, **p<0.01.

Mentions: To explore whether our ischemic diabetic wound showed an excessive inflammatory response, normally found in diabetic ischemic chronic wounds, we first detected wound neutrophils and macrophages infiltration by labeling MPO and CD68, specific enzyme or marker for these two inflammatory cells. Larger amount MPO expression cells were found on day 5 ischemic wounds, suggesting more neutrophils infiltration induced by ischemia (Fig 5A). More macrophages were also found in the ischemic diabetic wounds. Furthermore, macrophage resolution was also impaired, shown by the existing of much more CD68 positive cells on wounds at day 11 after wounding (Fig 5B).


Compromised Wound Healing in Ischemic Type 2 Diabetic Rats.

Yang P, Pei Q, Yu T, Chang Q, Wang D, Gao M, Zhang X, Liu Y - PLoS ONE (2016)

Ischemia induced exorbitant wound inflammatory response.(A) Representative images of MPO immunochemistry staining for both non-ischemic and ischemic wounds at day 5 after wounding. Scale bars = 200μm. (B) Representative images of immunohischemical staining for CD68, a specific marker of macrophage. Scale bars = 200μm. (C-F) Real time PCR analyze TNF-α, IL-6, IL-1β and IL-10 RNA expression (fold change) of non-ischemic and ischemic wounds at day 5 and 11 after wounding. Data was obtained from 12 wounds (n = 6), and are shown as the mean ± SD. **p<0.01. (G-I) Western blot analyze of wounds TNF-α, IL-6 and IL-10 at day 5 and 11 after wounding. Data was obtained from 12 wounds (n = 6), and are shown as the mean ± SD. *P<0.05, **p<0.01.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4814123&req=5

pone.0152068.g005: Ischemia induced exorbitant wound inflammatory response.(A) Representative images of MPO immunochemistry staining for both non-ischemic and ischemic wounds at day 5 after wounding. Scale bars = 200μm. (B) Representative images of immunohischemical staining for CD68, a specific marker of macrophage. Scale bars = 200μm. (C-F) Real time PCR analyze TNF-α, IL-6, IL-1β and IL-10 RNA expression (fold change) of non-ischemic and ischemic wounds at day 5 and 11 after wounding. Data was obtained from 12 wounds (n = 6), and are shown as the mean ± SD. **p<0.01. (G-I) Western blot analyze of wounds TNF-α, IL-6 and IL-10 at day 5 and 11 after wounding. Data was obtained from 12 wounds (n = 6), and are shown as the mean ± SD. *P<0.05, **p<0.01.
Mentions: To explore whether our ischemic diabetic wound showed an excessive inflammatory response, normally found in diabetic ischemic chronic wounds, we first detected wound neutrophils and macrophages infiltration by labeling MPO and CD68, specific enzyme or marker for these two inflammatory cells. Larger amount MPO expression cells were found on day 5 ischemic wounds, suggesting more neutrophils infiltration induced by ischemia (Fig 5A). More macrophages were also found in the ischemic diabetic wounds. Furthermore, macrophage resolution was also impaired, shown by the existing of much more CD68 positive cells on wounds at day 11 after wounding (Fig 5B).

Bottom Line: A bipedicle flap, with length to width ratio 1.5, was performed on the back of the rat to make the flap area ischemic.Our results demonstrated that a combination of HFD feeding and a low dose of STZ is capable of inducing the rats to develop type 2 diabetes with noticeable insulin resistance, persistent hyperglycemia, moderate degree of insulinemia, as well as high serum cholesterol and high triglyceride levels.Our study suggested that HFD feeding combined with STZ injection could induce type 2 diabetes in rat.

View Article: PubMed Central - PubMed

Affiliation: Department of Burn and Plastic Surgery, Shanghai Jiao Tong University School of Medicine, Ruijin Hospital, Shanghai 200025, China.

ABSTRACT
Ischemia is one of the main epidemic factors and characteristics of diabetic chronic wounds, and exerts a profound effect on wound healing. To explore the mechanism of and the cure for diabetic impaired wound healing, we established a type 2 diabetic rat model. We used an 8 weeks high fat diet (HFD) feeding regimen followed by multiple injections of streptozotocin (STZ) at a dose of 10mg/kg to induce Wister rat to develop type 2 diabetes. Metabolic characteristics were assessed at the 5th week after the STZ injections to confirm the establishment of diabetes mellitus on the rodent model. A bipedicle flap, with length to width ratio 1.5, was performed on the back of the rat to make the flap area ischemic. Closure of excisional wounds on this bipedicle flap and related physiological and pathological changes were studied using histological, immunohistochemical, real time PCR and protein immunoblot approaches. Our results demonstrated that a combination of HFD feeding and a low dose of STZ is capable of inducing the rats to develop type 2 diabetes with noticeable insulin resistance, persistent hyperglycemia, moderate degree of insulinemia, as well as high serum cholesterol and high triglyceride levels. The excision wounds on the ischemic double pedicle flap showed deteriorative healing features comparing with non-ischemic diabetic wounds, including: delayed healing, exorbitant wound inflammatory response, excessive and prolonged ROS production and excessive production of MMPs. Our study suggested that HFD feeding combined with STZ injection could induce type 2 diabetes in rat. Our ischemic diabetic wound model is suitable for the investigation of human diabetic related wound repair; especically for diabetic chronic wounds.

Show MeSH
Related in: MedlinePlus