DOI: 10.1556/2060.103.2016.4.5

Abstract: The mediodorsal prefrontal cortex (mdPFC) is a key structure of the central glucose-monitoring (GM) neural network. Previous studies indicate that intracerebral streptozotocin (STZ) microinjection-induced destruction of local chemosensory neurons results in feeding and metabolic alterations. The present experiments aimed to examine whether STZ microinjection into the mdPFC causes metabolic deficits. To do so, glucose tolerance test (GTT) and measurements of plasma metabolites were performed in STZ-treated or control rats. Intraperitoneal D-glucose load was delivered 20 min or 4 weeks following the intracerebral microinjection of STZ or saline (acute or subacute GTT, respectively). The STZ-treated rats displayed acute glucose intolerance: at the 120th min of the test, blood glucose level of these rats was significantly higher than that of the ones in the control group. When determining the plasma level of various metabolites, 30 min following the intracerebral STZ or saline microinjection, the triglyceride concentration of the STZ-treated rats was found to be reduced compared with that of the control rats. The GM neurons of the mdPFC are suggested to be involved in the organization of complex metabolic processes by which these chemosensory cells contribute to adaptive control mechanisms of the maintenance of homeostasis.

Keywords: forebrain limbic circuitry, mediodorsal prefrontal cortex, glucose-monitoring neurons, STZ, glucose intolerance, hypotriglyceridemia

DOI: 10.1556/2060.103.2016.4.001

Abstract: Diabetic nephropathy (DN) is one of the most common microvascular diabetic complications. This study was designed to evaluate the possible protective effect and underlying mechanisms of HO-1 induction in streptozotocin (STZ)-induced early DN in rats. The diabetic rats were divided into three groups: STZ-diabetic, cobalt protoporphyrin (CoPP)-treated diabetic, and zinc protoporphyrin IX (ZnPP)-treated diabetic groups. Compared to the STZdiabetic group, CoPP-induced HO-1 upregulation improved the diabetic state and renal functional parameters, suppressed the renal proinflammatory marker, NF-κB, abrogated the elevated renal hydroxyprolin, and decreased the enhanced renal nicotinamide adenine dinucleotide phosphate oxidase activity with parallel reduction of urinary oxidative stress markers. On the contrary, treatment with ZnPP abrogated HO-1 levels, aggravated the diabetic condition with further increases in renal oxidative stress, fibrotic and inflammatory markers, and exacerbated renal dysfunction in diabetic animals. These findings suggest that the reduced diabetic renal injury upon HO-1 induction implicates the role of HO-1 induction as a potential treatment for DN.

Keywords: diabetic nephropathy, heme oxygenase, cobalt protoporphyrin, zinc protoporphyrin IX, renal hydroxyprolin

DOI: 10.1556/2060.103.2016.4.3

Abstract: This study aimed to assess the role of H2S in homocysteine-induced cardiodynamic effects in the isolated rat heart. The hearts were retrogradely perfused according to the Langendorff technique. The maximum and minimum rates of pressure in the left ventricle (dp/dt max, dp/dt min), systolic and diastolic left ventricular pressures (SLVP, DLVP), heart rate (HR), and coronary flow (CF) were measured. A spectrophotometrical method was used to measure the following oxidative stress markers: index of lipid peroxidation (thiobarbituric acid reactive substances, TBARS), nitrite level (NO2 −), superoxide anion radicals (O2 •−), and hydrogen peroxide (H2O2) concentrations. The administration of 10 μmol/l DL-homocysteine (DL-Hcy) alone decreased dp/dt max, SLVP, and CF but did not change any oxidative stress parameters. The administration of 10 μmol/l DL-propargylglycine (DL-PAG) decreased all cardiodynamic parameters and increased the concentration of O2 •−. The co-administration of DL-Hcy and DL-PAG induced a significant decrease in all estimated cardiodynamic parameters and decreased the concentration of NO2 − and O2 •− but increased the levels of TBARS and H2O2. Homocysteine shows a lower pro-oxidative effect in the presence of hydrogen sulfide (H2S), which indicates a potential anti-oxidative capacity of H2S.

Keywords: cardiodynamics, homocysteine, H2S, Langendorff technique, oxidative stress

DOI: 10.1556/2060.103.2016.4.4

Abstract: Quinones are among the rare compounds successfully used as therapeutic agents to correct mitochondrial diseases and as specific regulators of mitochondrial function within cells. The aim of the present study was to elucidate the redox-dependent effects of quinones on mitochondrial function. The functional parameters [respiratory activity, membrane potential, and reactive oxygen species (ROS) generation] of isolated rat liver mitochondria and mitochondria in intact cells were measured in the presence of eight exogenously applied quinones that differ in lipophilicity and one-electron reduction potential. The quinones affected the respiratory parameters of mitochondria, and dissipated the mitochondrial membrane potential as well as influenced (either decreased or enhanced) ROS generation, and restored the electron flow during electron transport chain inhibition. The stimulation of ROS production by juglone and 2,5-di-tert-butyl-1,4-benzoquinone was accompanied by a decrease in the acceptor control and respiration control ratios, dissipation of the mitochondrial membrane potential and induction of the reverse electron flow under succinate oxidation in isolated mitochondria. Menadione and 2,3,5-trimethyl-1,4-benzoquinone, which decreased the mitochondrial ROS generation, did not affect the mitochondrial potential and, vice versa, were capable of restoring electron transport during Complex I inhibition. In intact C6 cells, all the quinones, except for coenzyme Q10, decreased the mitochondrial membrane potential. Juglone, 1,4-benzoquinone, and menadione showed the most pronounced effects. These findings indicate that quinones with the reduction potential values E1/2 in the range from −99 to −260 mV were effective redox regulators of mitochondrial electron transport.

Keywords: quinone, mitochondria, respiration, membrane potential, reactive oxygen species, one-electron reduction potential

DOI: 10.1556/2060.103.2016.4.6

Abstract: Background: Oxidative stress plays a critical role in the pathogenesis and progression of type 2 diabetes and diabeticassociated cardiovascular complications. This study investigated the impact of crocin combined with voluntary exercise on heart oxidative stress indicator in high-fat diet-induced type 2 diabetic rats. Materials and methods: Rats were divided into four groups: diabetes, diabetic-crocin, diabetic-voluntary exercise, diabetic-crocin-voluntary exercise. Type 2 diabetes was induced by high-fat diet (4 weeks) and injection of streptozotocin (intraperitoneally, 35 mg/kg). Animals received crocin orally (50 mg/kg); voluntary exercise was performed alone or combined with crocin treatment for 8 weeks. Finally, malondialdehyde (MDA), activity of antioxidant enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) were measured spectrophotometrically. Results: Treatment of diabetic rats with crocin and exercise significantly decreased the levels of MDA (p < 0.001) and increased the activity of SOD, GPx, and CAT compared with the untreated diabetic group. In addition, combination of exercise and crocin amplified their effect on antioxidant levels in the heart tissue of type 2 diabetic rats. Conclusion: We suggest that a combination of crocin with voluntary exercise treatment may cause more beneficial effects in antioxidant defense system of heart tissues than the use of crocin or voluntary exercise alone.

Keywords: type 2 diabetes, crocin, voluntary exercise, antioxidant, oxidative stress

DOI: 10.1556/2060.103.2016.4.1

Abstract: Background: Type 2 diabetes mellitus (T2DM)-induced neuropathy and ischemia-reperfusion post-surgery prolong carpal tunnel syndrome (CTS) pathology, but the effect of T2DM on the prognostic outcome of carpal tunnel (CT) release surgery needs to be investigated. Materials and methods: A total of 64 individuals with CTS underwent CT release surgery. HbA1c levels identified their diabetic status. The individual prognostic outcomes were measured by nerve conduction velocity (NCV), amplitude, and latency. Measurement of [Ca2+]c and reactive oxygen species (ROS) from isolated endothelial cells (ECs) revealed the oxidative burden of the normal and diabetic CTS phenotypes. Results: CTS individuals with HbA1c >7 showed decreased NCV (≈22 m/s) and amplitude (≈4.2 mV) with increased latency (≈6 ms), compared to groups with HbA1c ≤ 7. Further to CT release surgery, the reversal of the nerve conduction to normalcy was greatly influenced by the diabetic profile of the individuals. Our results showed elevated basal [Ca2+]c and corresponding high cytosolic ROS in the ECs isolated from individuals with HbA1c > 7 compared to the diabetic and healthy control groups. Conclusion: The individuals with diabetic index showed suboptimal neuronal performance pre- and post-CT release surgery. Oxidative stress mediated by high [Ca2+]c and ROS of ECs dissipates to adjoining cells worsening the pathology of the untreated CTS.

Keywords: carpal tunnel syndrome, diabetic CTS, endothelial calcium, endothelial ROS, post surgery prolonged CTS, oxidative stress, T2DM complications

DOI: 10.1556/2060.103.2016.4.2

Abstract: The effects of moderate hypothermia (28 °C) on the response of human varicose spermatic vein to α1-adrenoceptor agonist phenylephrine and the role of endothelial nitric oxide (NO) in these effects were studied. Concentration– response curves for phenylephrine (10−9 to 3 × 10−4 M) were recorded in rings with and without endothelium at 37 and 28 °C. To further analyze the role of NO, in the response to phenylephrine during hypothermia, the effects of this agonist in the presence of NG-nitro-L-arginine methyl ester (10−4 M) were also determined. Under every condition tested, phenylephrine produced a marked, concentration-dependent contraction. Sensitivity of intact veins to the agonist was consistently lower at 28 °C than at 37 °C. There was no significant difference in phenylephrine response at 28 and 37 °C in vessels without endothelium but at 28 °C veins without endothelium showed a higher sensitivity than intact veins to phenylephrine. The sensitivity of veins with and without endothelium to nitroprusside (10−9 to 3 × 10−3 M) was significantly decreased during hypothermia, and endothelium removal did not affect the relaxation to this nitrovasodilator. These results suggest that moderate hypothermia decreases the sensitivity of human varicose spermatic vein to phenylephrine probably by increasing the availability of endothelial NO.

Keywords: endothelium, hypothermia, NO, phenylephrine, spermatic vein, temperature

DOI: 10.1556/2060.103.2016.4.002