sodium-nitrite and daidzein

Endothelial nitric-oxide synthase (eNOS) acts as a common pathogenic pathway in diabetic nephropathy (DN). However, its functional consequences are still not fully understood. Caveolin, a membrane protein, inhibits the eNOS by making caveolin-eNOS complex, and its expression is upregulated during diabetes mellitus (DM). This study was designed to determine the role of caveolin in eNOS-mediated NO synthesis and release in DN. DM in rat was induced by feeding of high-fat diet (HFD) for 2 weeks, followed by single dose of streptozotocin (STZ) (35 mg/kg, ip) further followed by HFD for further 8 weeks. Serum nitrite/nitrate ratio was measured to determine the plasma level of NO. Diabetic rat, after 6 weeks of STZ, developed elevated level of BUN, protein in urine, urinary output, serum creatinine, serum cholesterol, kidney weight, kidney weight/body weight, and renal cortical collagen content, while serum nitrite/nitrate concentration was significantly decreased as compared to normal control group. Treatment with sodium nitrite (NO donor), L: -arginine (NO precursor), daidzein (caveolin inhibitor), and combination of L: -arginine and daidzein for 2 weeks markedly attenuated these changes and increased serum nitrite/nitrate ratio. However, treatment with L-NAME, a eNOS inhibitor, significantly attenuated the L: -arginine-, daidzein-, or combination of L: -arginine and daidzein-induced ameliorative effects in DN. The finding of this study suggests that caveolin plays a vital role in the eNOS-mediated decrease in renal level of NO, which may be responsible for the development of DN in rats.

Topics: Animals; Arginine; Blood Glucose; Blood Urea Nitrogen; Body Weight; Caveolin 1; Cholesterol; Collagen; Creatinine; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Dietary Fats; Endothelium, Vascular; Isoflavones; Kidney; Male; Nitric Oxide Synthase Type III; Nitrites; Organ Size; Proteinuria; Rats; Rats, Wistar; Sodium Nitrite; Urine

We have already found that nitrite-treated isoflavones exhibit genotoxic activities toward Salmonella typhimurium TA 100 and 98 strains (submitted: nitrite-treated genistein). However, we have not demonstrated genotoxic activity induced by simultaneous treatment with isoflavones and NaNO(2)in vivo. In the present study, we examined whether coadministration of isoflavones (such as daidzein and genistein) and NaNO(2) induces DNA damage in the stomach of ICR male mice. Mice were coadministered with isoflavones (1mg/kg body weight) and NaNO(2) (10mg/kg body weight), and dissected to collect tissues at 1, 3, and 6h after administration. We used comet assay combined with repair enzyme formamidopyrimidine-N-glycosylase (FPG) to detect FPG-sensitive sites. An HPLC-ECD system was employed to determine 8-oxo-2'-deoxyguanosine (8-oxodG) in the stomach. In addition, we observed leukocyte infiltration by histopathological investigation, and measured total superoxide dismutase (SOD) in the stomach. We confirmed that oxidative DNA damage in the stomach was significantly increased by coadministration. Total SOD activities were also significantly stimulated by coadministration. However, the induction of inflammation in the stomach was not found. These data suggest that coadministration of isoflavones and NaNO(2) can cause DNA damage in the stomach because of the formation of radicals.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Animals; Comet Assay; Deoxyguanosine; DNA; DNA Damage; Genistein; Glycine max; Isoflavones; Male; Mice; Mice, Inbred ICR; Neutrophil Infiltration; Sodium Nitrite; Stomach; Superoxide Dismutase