Nitric Oxide Synthase in the Skeletal Muscle and Arterioles of Rats with Streptozotosin-Induced Diabetes Mellitus

Authors: CİHAN DEMİRCİ, KADRİYE AKGÜN DAR, İLHAN UYANER

Abstract: Nitric oxide synthase (NOS) that uses NADPH as a cofactor is an enzyme which produces nitric oxide (NO) from L-arginine. Endothelial-derived (eNOS), inducible (iNOS) and neuronal (nNOS) nitric oxide synthase are 3 known isoforms. Endothelial NO regulates the vascular tonus and causes the dilatation of vessels, while NOS localized in the sarcolemma of striated muscle plays a role in the regulation of muscle hemodynamics. Many authors have found differences in NO production in some pathological conditions such as diabetes mellitus (DM), hypertension and atherosclerosis. In this study, NADPH-diaphorase (NADPH-d) was histochemically employed to explore any changes in NO production associated with the degree of DM, by detecting the enzyme in the muscle tissue. Diabetes was induced by injecting Swiss albino rats with 65mg/kg of streptozotosin (STZ). Two, 4, 6 and 12 weeks after the STZ injection, the spinotrapezius muscles of the animals were fixed in paraformaldehyde and NADPH-d histochemistry was applied to cryostat sections. Six and 12 weeks after DM, NOS increased both in the muscles and the endothelial cells of arterioles, being more prominent in the 12-week group in which the NADPH-d reaction was also localized in sarcoplasm together with the sarcolemma of muscle fibers than in controls. NOS was more abundant in the 2nd- and 3rd-order arterioles in diabetic animals than in controls. After the superfusion of the N^W-nitro-L-Arginine Methyl Ester (L-NMA) that inhibits NOS, the NADPH-d reaction was still present in 12-week DM, while not in controls. ACh superfusion caused endothelial dilation and increased NO production in both the control and diabetic groups. This increase was observed to be more pronounced in diabetics than in controls. We also superfused sodium nitroprusside as a NO donor, and found resulting dilation in arterioles, with differences between the control and DM groups, the latter being more conspicuous.

Keywords: Nitric oxide synthase (NOS), diabetes mellitus, skeletal muscle, arteriole

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