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Superoxide production pathways in aortas of diabetic rats: beneficial effects of insulin therapy and endurance training

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Abstract

Superoxide (O ·−2 ) overproduction, by decreasing the nitric oxide (·NO) bioavailability, contributes to vascular complications in type 1 diabetes. In this disease, the vascular O ·−2 can be produced by the NADPH oxidase (NOX), nitric oxide synthase (NOS), and xanthine oxidase (XO). This study aimed to determine the contribution of each enzymatic pathway in hyperglycemia-induced O ·−2 overproduction, and the effects of an endurance training program and insulin therapy, associated or not, on the O ·−2 production (amount and related enzymes) in diabetic rats. Forty male Wistar rats were divided into diabetic (D), diabetic treated with insulin (D-Ins), diabetic trained (D-Tr), or diabetic insulin-treated and trained (D-Ins + Tr) groups. An additional healthy group was used as control. Insulin therapy (Glargine Lantus, Sanofi) and endurance training (treadmill run: 60 min/day, 25 m/min, 5 days/week) started 1 week after diabetes induction by streptozotocin (45 mg/kg), and lasted for 8 weeks. At the end of the protocol, the O ·−2 production in aorta rings was evaluated by histochemical analyses (DHE staining). Each production pathway was studied by inhibiting NOX (apocynin), NOS (L-Name), or XO (allopurinol) before DHE staining. Diabetic rats exhibited hyperglycemia-induced O ·−2 overproduction, resulting from NOX, NOS, and XO activation. Insulin therapy and endurance training, associated or not, decreased efficiently and similarly the O ·−2 overproduction. Insulin therapy reduced the hyperglycemia and decreased the three enzymatic pathways implicated in the O ·−2 production. Endurance training decreased directly the NOS and XO activity. While both therapeutic strategies activated different pathways, their association did not reduce the O ·−2 overproduction more significantly.

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Notes

  1. Among the two types of NOS isoforms in aorta, uncoupled eNOS has been reported as the major source of superoxide anion production, comparative to iNOS (negligible). Hereinafter, L-Name was considered to essentially inhibit superoxide anion production by eNOS uncoupling.

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Acknowledgments

The authors thank the platform of fluorescence microscopy (H2P2, Rennes), Pascale Bellaud and Roselyne Viel for their technical support.

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The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Laboratoire M2S, Université Rennes 2 – ENS Cachan, Avenue Robert Schuman, 35170, Bruz, France

    L. Malardé, A. Rebillard, S. Le Douairon-Lahaye, S. Vincent, M. S. Zguira, S. Lemoine-Morel, A. Gratas-Delamarche & C. Groussard

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  1. L. Malardé
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  2. A. Rebillard
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  3. S. Le Douairon-Lahaye
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  4. S. Vincent
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  5. M. S. Zguira
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  6. S. Lemoine-Morel
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  7. A. Gratas-Delamarche
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  8. C. Groussard
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Correspondence to L. Malardé.

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Malardé, L., Rebillard, A., Le Douairon-Lahaye, S. et al. Superoxide production pathways in aortas of diabetic rats: beneficial effects of insulin therapy and endurance training. Mol Cell Biochem 389, 113–118 (2014). https://doi.org/10.1007/s11010-013-1932-z

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  • DOI: https://doi.org/10.1007/s11010-013-1932-z

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