Jana Vlkovicova

Rosmarinic acid administration attenuates diabetes-induced vascular dysfunction of the rat aorta

Oxidative stress as well as inflammation processes are engaged in diabetic vascular complications. Rosmarinic acid, a natural phenol antioxidant carboxylic acid, was found to have multiple biological activity, including anti‐inflammatory and antitumour effects, which are a consequence of its inhibition of the inflammatory processes and of reactive oxygen species scavenging. The aim of this work was to study effects of rosmarinic acid administration on vascular impairment induced by experimental diabetes in rats.

Supplementation with n-3 polyunsaturated fatty acids to lipopolysaccharide-induced rats improved inflammation and functional properties of renal Na,K-ATPase

Measurements of enzyme kinetics of renal Na, K-ATPase were used for characterization of ATP- and Na⁺-binding sites in rats that were subjected to 10 days of moderate inflammation that was induced by a single dose of Escherichia coli lipopolysaccharides (LPSs) at a dose of 1 mg kg⁻¹ body weight. We hypothesized that LPSs might initiate a malfunction of renal Na, K-ATPase, which is a key enzyme involved in regulation of sodium homeostasis in the organism. We also investigated the potential effect that fish oil (FO) has in the prevention of Na, K-ATPase alterations by administering FO daily at a dose of 30 mg kg⁻¹. Alone, LPS elevated the level of C-reactive protein by more than 500% and free radicals by 36% in plasma, as indicated by an increased level of malondialdehyde. The Na, K-ATPase was slightly altered in the vicinity of the ATP-binding site as suggested by the 9% increase of the concentration of ATP necessary for half-maximal activation of the enzyme, thus indicating a deteriorated binding of ATP as a consequence of inflammation. Daily supplementation of FO partly attenuated LPS-induced injury, as observed by a significant decrease in the plasma levels of C-reactive protein and free radicals, hence maintaining the activity of renal Na, K-ATPase to the level of healthy control animals. In conclusion, our findings showed that FO prevented an excessive malondialdehyde production in LPS-treated animals and stabilized renal Na, K-ATPase.

EFFECT OF MATURATION ON RENAL Na+/K+-ATPase AND ITS SUSCEPTIBILITY TO NITRIC OXIDE-DEFICIENT HYPERTENSION IN RATS

1 The present study deals with the effect of maturation on the kinetic properties of renal Na+/K+‐ATPase and its susceptibility to nitric oxide (NO)‐deficient hypertension induced by the NO synthase inhibitor NG‐nitro‐l‐arginine methyl ester (l‐NAME). 2 Immature (4‐week‐old) and adult (12‐week‐old) male Wistar rats were administered l‐NAME (40 mg/kg per day) in their drinking water for 4 weeks. 3 The properties of the ATP‐ and Na+‐binding sites of Na+/K+‐ATPase were investigated by activation of the enzyme with increasing concentrations of the energy substrate ATP and/or cofactor Na+. Unchanged values of Km suggest that energy utilization by the enzyme in the kidney of control rats remains unaffected during maturation. Conversely, the decrease in KNa values (the concentration of Na+ necessary to achieve half‐maximal reaction velocity) indicates improved affinity for Na+ in the older group of control rats. 4 Application of l‐NAME to all young animals had no significant effect on the functional properties of Na+/K+‐ATPase. 5 In adult animals, the Vmax values remained unchanged after treatment with l‐NAME, but the affinities of the ATP‐ and Na+‐binding sites were decreased, as indicated by significant increase in Km and KNa values. 6 Maturation of control rats was accompanied by an increase in the Na+ affinity of renal Na+/K+‐ATPase without affecting ATP utilization. However, maturation increased the susceptibility of renal Na+/K+‐ATPase to the harmful effects of l‐NAME.

Dual effect of polyphenolic compounds on cardiac Na+/K+-ATPase during development and persistence of hypertension in rats.

The enzyme kinetics of cardiac Na(+)/K(+)-ATPase were used for characterizing the ATP- and Na(+)-binding sites after administration of red wine polyphenolic compounds (Provinol) during developing and sustained hypertension. Hypertension was induced in rats (LN group) by the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME, 40 mg*kg(-1)*day(-1)). Provinol (40 mg*kg(-1)*day(-1)) was applied during developing hypertension (LNPF4 group) and sustained hypertension (LNPF7/3 group). Provinol reduced the number of active Na(+)/K(+)-ATPase molecules in cardiac tissue, as indicated by decreased V(max) values (by 33% in LNPF4 and 26% in LNPF7/3 compared with LN). Concerning qualitative properties of the enzyme, Provinol induced different effects on the ATP- and Na(+)-binding sites of Na(+)/K(+)-ATPase. The ATP-binding site was impaired by Provinol, as indicated by increased K(m) value (by 52% in LNPF4 vs. LN), suggesting worsened utilization of substrate by the enzyme. In sustained hypertension, however, Provinol had no effect on the ATP-binding site, as indicated by unchanged K(m) value (LNPF7/3 vs. LN). On the other hand, the Na(+)-binding site was protected by Provinol, as suggested by decreased K(Na) value (by 72% in LNPF4 and 69% in LNPF7/3 vs. LN), indicating an increased affinity of the enzyme for sodium. Thus, Provinol appeared to stimulate the extrusion of Na(+) from cardiac cells, especially in the physiologically important range of sodium concentrations (2-10 mmol*L(-1)), during both developing and sustained hypertension.

Sex differences in endothelial function of aged hypertriglyceridemic rats – effect of atorvastatin treatment

ABSTRACT The aim of the study was to test the hypothesis that the effect of atorvastatin on endothelium-dependent relaxation of the superior mesenteric artery (SMA) may differ in male vs. female aged hypertriglyceridemic rats (HTGs). Experiments were performed on 11-month-old male and female Prague hereditary HTGs. Atorvastatin (ATO) was administered p.o. in the dose of 0.30 mg/100g/day. Controls received vehiculum. After two months of ATO administration blood pressure, serum triglycerides (TG) and total cholesterol (CHOL) were determined. Endothelial function of SMA was studied in vitro using evaluation of relaxant responses of precontracted SMA to acetylcholine. The serum TG of control male HTGs were found to be statistically higher than those of female controls, while CHOL and blood pressure did not share gender differences. Responses of SMA of female control HTGs were statistically decreased compared to their male counterparts. ATO treatment induced decrease in blood pressure and TG of both males and females, yet CHOL values were reduced only in females. The protective effect of ATO on SMA endothelial function was much more pronounced in females compared to males. We conclude that vascular endothelial dysfunction of aged HTG rats is more severe and more attenuated by ATO in females compared to males. The protective effect of ATO on vascular endothelial function does not seem to depend solely on its lipid lowering action.