Our previous study demonstrated that an impaired sonic hedgehog (Shh) pathway contributed to cardiac dysfunction in type 1 diabetic mice with
myocardial infarction (MI). The present study aimed to test the hypothesis that oxidative stress may contribute to the impaired Shh pathway and cardiac dysfunction in type 1 diabetic mice with MI.
Streptozotocin-induced type 1 diabetic mice (C57/Bl6, male) and rat neonatal cardiomyocytes were used in the present study. Mice were randomly assigned to undergo
ligation of the coronary artery or pseudosurgery. A potent
antioxidant Tempol was administered in vivo and in vitro. Cardiac function was assessed by echocardiography, capillary density by immunohistochemisty, percentage of
myocardial infarct using Massons trichrome staining,
reactive oxygen species detection using
dihydroethidium dye or 2,7-dichlorofluorescein diacetate probe and
protein expression levels of the Shh pathway by western blot analysis. The
antioxidant Tempol was shown to significantly increase myocardial
protein expression levels of Shh and patched-1 (Ptc1) at 7-18 weeks and improved cardiac function at 18 weeks in type 1 diabetic mice, as compared with mice receiving no
drug treatment. Furthermore, myocardial
protein expression levels of Shh and Ptc1 were significantly upregulated on day 7 after MI, and capillary density was enhanced. In addition, the percentage area of
myocardial infarct was reduced, and the cardiac dysfunction and survival rate were improved on day 21 in diabetic mice treated with
Tempol. In vitro, treatment of rat neonatal cardiomyocytes with a mixture of
xanthine oxidase and
xanthine decreased
protein expression levels of Shh and Ptc1 in a concentration-dependent manner, and
Tempol attenuated this effect. These results indicate that oxidative stress may contribute to an impaired Shh pathway in type 1 diabetic mice, leading to diminished myocardial healing and cardiac dysfunction. Antioxidative strategies aimed at restoring the endogenous Shh pathway may offer a useful means for improving diabetic cardiac function.