The effects of recombinant human erythropoietin (rHuEPO) in diabetes-related healing defects were investigated by using an incisional skin-wound model produced on the back of female diabetic C57BL/KsJm ؉/؉ Lept db mice (db ϩ /db ϩ ) and their normoglycemic littermates (db ϩ/ϩ m). Animals were treated with rHuEPO (400 units/kg in 100 l s.c.) or its vehicle alone (100 l). Mice were killed on different days (3, 6, and 12 days after skin injury) for measurement of vascular endothelial growth factor (VEGF) mRNA expression and protein synthesis, for monitoring angiogenesis by CD31 expression, and for evaluating histological changes. Furthermore, we evaluated wound-breaking strength at day 12. At day 6, rHuEPO injection in diabetic mice resulted in an increase in VEGF mRNA expression (vehicle ‫؍‬ 0.33 ؎ 0.1 relative amount of mRNA; rHuEPO ‫؍‬ 0.9 ؎ 0.09 relative amount of mRNA; P < 0.05) and protein wound content (vehicle ‫؍‬ 23 ؎ 5 pg/wound; rHuEPO ‫؍‬ 92 ؎ 12 pg/wound; P < 0.05) and caused a marked increase in CD31 gene expression (vehicle ‫؍‬ 0.18 ؎ 0.05 relative amount of mRNA; rHuEPO ‫؍‬ 0.98 ؎ 0.21 relative amount of mRNA; P < 0.05) and protein synthesis. Furthermore, rHuEPO injection improved the impaired wound healing and, at day 12, increased the wound-breaking strength in diabetic mice (vehicle ‫؍‬ 12 ؎ 2 g/mm; rHuEPO 21 ؎ 5 g/mm; P < 0.05). Erythropoietin may have a potential application in diabetes-related wound disorders.

Duchenne muscular dystrophy (DMD) is a progressive muscle-wasting disease resulting from lack of the sarcolemmal protein dystrophin. However, the mechanism leading to the final disease status is not fully understood. Several lines of evidence suggest a role for nuclear factor (NF)-B in muscle degeneration as well as regeneration in DMD patients and mdx mice. We investigated the effects of blocking NF-B by inhibition of oxidative stress/lipid peroxidation on the dystrophic process in mdx mice. Five-week-old mdx mice received three times a week for 5 weeks either IRFI-042 (20 mg/kg), a strong antioxidant and lipid peroxidation inhibitor, or its vehicle. IRFI-042 treatment increased forelimb strength (؉22%, P < 0.05) and strength normalized to weight (؉23%, P < 0.05) and decreased fatigue (؊45%, P < 0.05). It also reduced serum creatine kinase levels (P < 0.01) and reduced muscle-conjugated diene content and augmented muscle-reduced glutathione (P < 0.01). IRFI-042 blunted NF-B DNA-binding activity and tumor necrosis factor-␣ expression in the dystrophic muscles (P < 0.01), reducing muscle necrosis (P < 0.01) and enhancing regeneration (P < 0.05). Our data suggest that oxidative stress/lipid peroxidation represents one of the mechanisms activating NF-B and the consequent pathogenetic cascade in mdx muscles.

SUMMARY:Nuclear factor (NF)-B plays a central role in acute pancreatitis. We studied cerulein (CER)-induced pancreatitis in NF-B knockout (KO) mice. NF-B KO mice and normal control littermate wild-type (WT) mice were given four hyperstimulating doses of cerulein every hour to elicit secreatagogue-induced pancreatitis. Malonildialdehyde activity, glutathione levels, myeloperoxidase activity, TNF-␣, and NF-B binding activity and its inhibitory protein IB␣ were studied in the pancreas. Furthermore, we measured plasma lipase and amylase and the histological damage. KO mice had reduced malonildialdehyde levels (WT ϩ CER ϭ 4.083 Ϯ 0.95 mol/g; KO ϩ CER ϭ 1.513 Ϯ 0.63 mol/g), decreased myeloperoxidase activity (WT ϩ CER ϭ 19.3 Ϯ 2.39 mU/g; KO ϩ CER ϭ 10.21 Ϯ 2.05 mU/g), increased glutathione levels (WT ϩ CER 6.22 Ϯ 2.46 mol/g; KO ϩ CER ϭ 15. 516 Ϯ 2.92 mol/g), and reduced serum levels of amylase (WT ϩ CER ϭ 2519 Ϯ 656.9 U/L; KO ϩ CER ϭ 916 Ϯ 280.4 U/L) and lipase (WT ϩ CER ϭ 1420 Ϯ 170 U/L; KO ϩ CER ϭ 861 Ϯ 172. 3 U/L). KO mice showed reduced pancreatic NF-B activation, decreased TNF-␣ tissue content, and reduced histologic alterations. Our data suggest that KO mice have an attenuated cerulein-induced pancreatitis and help to define the possible interaction between NF-B activation and oxidative stress in this deleterious event. (Lab Invest 2003, 83:1723-1732.

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These data suggest that IRFI 042 blocks the activation of NF-kappaB, reduces TNF-alpha mRNA levels, and finally reverses endotoxic shock.