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- În acest studiu, demonstrăm că miR de șoarece este localizat predominant în nucleul diferitelor tipuri de celule și că modelul său de localizare nucleară se schimbă rapid pe stimuli apoptotici.
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PDF, 1. MET improved global activity and increased the time spent experimento biogenesis anti aging unprotected open arms, decreased oxidative stress, both in the frontal lobe and in the hippocampus, and increased neuroglobin expression in the frontal cortex. Thus, our findings suggest that metformin has antioxidant and anxiolytic effects and may partially reverse the neurotoxic effects induced by ethanol.
Dictionar Tehnic Roman Englez
Keywords Alcohol. Oxidative stress. Patients frequently reach the dependence stage after self-treating anxiety and depressive feelings with alcohol Rehm et al. However, the same symptoms; can be secondary to alcohol withdrawal Conway et al. Chronic ethanol exposure produces neurotoxicity by affecting synaptic plasticity and the function of glial cells, but also causes neuroinflammation and neuronal death by necrosis, apoptosis, and oxidative stress Volkow et al.
In addition, chronic alcohol administration impairs the synaptic formation and synaptic plasticity, learning, and memory due to involvement of the brain extracellular matrix ECM with an important role in behavioral responses to drugs of abuse and alcohol 4. Metformin METthe most effective treatment for type2 diabetes T2Dis an oral antidiabetic drug that inhibits hepatic glucose production, with a dimethyl-biguanide structure, that was synthetized in from the European Galega officinalis plant French lilac Thomas and Gregg Additionally, it has a protective effect against T2D complications, such as diabetic angiopathy and nephropathy and an antidepressant effect by improving cognitive functions in diabetic depressed patients Guo et al.
Furthermore, MET was investigated in many other diseases, such as neurocognitive disorders, cerebral ischemia, cardiovascular disease, obesity, neoplasms, and polycystic ovarian syndrome.
MET is also used for the treatment of antipsychotic weight gain, as an antibiotic, active on gut experimento biogenesis anti aging, and as an anti-aging drug Arbelaez-Quintero and Palacios ; Balen et al. Its exact mechanism of action is not yet fully understood, thus numerous pathways were proposed. The most accepted targets are the activation of AMP-activated protein kinase AMPKleading to a catabolic state with the inhibition of ATP-consuming processes, such as glucose, protein, and lipid synthesis Lage et al.
MET experimento biogenesis anti aging shown to protect endothelial cells from apoptosis during hyperglicemia by inhibiting PTP experimento biogenesis anti aging and cytochrome-c release and by restoring the vascular endothelial growth factor VEGF A pathway Bakhashab et al.
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Another mechanism of action of MET is the reduction of reactive oxygen species production with diminished oxidative stress and reduced DNA damage Algire et al. A recent paper concluded that metformin stimulates neurogenesis in the adult brain, by experimento biogenesis anti aging the production of new neurons in the olfactory bulb and the dentate gyrus of the hippocampus, important regions for emotions and memory Wang et al.
Physiologically, adult neurogenesis is a phenomenon present in the dentate gyrus of the hippocampus, the sub-ventricular ultra corection line repair soin yeux anti ride crema antirid chanel, and in the olfactory bulb throughout the entire life Ahmed et al. All other chemicals and reagents were purchased from Sigma-Aldrich Chemicals GmbH and were of high-grade purity.
The animals were kept under standard laboratory conditions h light dark cycle, 24 ± 2 °Cand had free access to a standard pellet diet and water. The frontal cortex and hippocampus from 4 rats in each group was collected for histopathological testing.
From the other 6 rats of every group, the frontal cortex and hippocampus were harvested for oxidative stress evaluation and ELISA testing. The dose for the MET and the time point for all the assays that were performed were considered regarding other studies which had found a protective effect of MET for memory impairment in Wistar rats Ghadernezhad et al.
Alcohol dose was also established with regard to other studies that evaluated hippocampal neurodegeneration in rats Motaghinejad et al. Behavioral Testing Two different tests were used in this particular study, such as the OFT and the EPM to assess the general locomotor activity experimento biogenesis anti aging anxiety-like behavior.
OFT Rats were placed in the center of an open field arena × × 40 cm that was electronically divided into 9 equal squares. The animals were freely allowed to explore the apparatus for 5 min.
Total and peripheral traveled distance and total and peripheral numbers of entries were considered as a general locomotion index. EPM The apparatus consists of a plus-shaped maze, with two open 10 × 50 cm and two closed 10 × 50 × 40 cm arms, elevated to 60 cm aboveground level. Measures of motor activity in this task are total and closed arm traveled distance and total and closed arm number of made entries.
Oxidative and Nitrosative Stress Assessment Malondialdehyde Malondialdehyde MDA levels were used to assess lipid peroxidation in the experimento biogenesis anti aging and frontal lobes and were determined using the fluorimetric method with 2-thiobarbituric acid TBA described by Conti et al.
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The samples of skin homogenate were heated on a boiling water bath for 1 h with a solution of 10 mM 2-thiobarbituric acid in 75 mM K2HPO4 pH 3. After cooling, the reaction product was extracted with 5 ml n-butanol. The MDA concentration was determined in the organic phase excitation at nm and emission at nm.
Nitrotyrosine Nitrotyrosine levels were considered as a nitrosative stress parameter. Nitric Oxide Nitric oxide NO levels in the hippocampus and in the frontal lobes were measured by Griess reaction for nitrite plus nitrate quantification using a two-step procedure.
First, the nitrate in the samples was reduced to nitrite in a conversion buffer containing glucosephosphate 2.
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After incubation, the proteins were precipitated with 3. The samples absorbance was read at nm. Catalase Catalase activity in the homogenates and lysates was measured in a reaction mixture containing 10 mM hydrogen peroxide in 50 mM kalium phosphate buffer, pH 7. The enzyme quantity which produced an absorbance reduction of 0.
Their endogenous inhibitor is the tissue inhibitor 1 of metalloproteinase TIPM After paraffin embedding, sections were cut at 5 μm and mounted on electrostatically charged glass slides. Tissue sections were dewaxed in xylene, rehydrated, and produs anti-imbatranire with Mayer hematoxylin-eosin and Nissl staining for histological exam.
The sections were then incubated at 4 °C for 12 h with rabbit monoclonal anti-neuroglobin antibody, Sigma HPA AB dilution The slides were washed with distilled water, counter-stained with hematoxylin for 3 min, and the excess removed with rinse in distilled water. After bluing in alkaline water for 5 min, 3 shorter baths of absolute ethanol, 1 bath of xylene were made, and then the sections were covered with synthetic resin Merck, Darmstadt, Germany.
Quantitative Estimation of Neuroglobin Expression Statistical Analysis Neuroglobin quantification was performed by the Western blot technique. Following the Biorad Miniprotean system BioRadthe lysates were transferred to polyvinylidenedifluoride membranes BioRad. Blots were blocked and incubated with Sigma-Aldrich neuroglobin antibodies diluted in MET administration improved total traveled distance by This test mainly assesses anxiety-like behavior high open arm traveled distance and open arms number of entries and time ratio—low level of anxietybut the locomotor activity can be recorded as well Sevastre-Berghian et al.
In experimento biogenesis anti aging EPM, the number of entries in closed arms Fig. Although the exploratory activity was decreased, in the EPM, the total traveled distance Fig. Table 1 Data for Fig. Each group consisted of 7 rats. Results are expressed as mean ± SEM. The EPM test was designed to assess the anxiety-like behavior of experimento biogenesis anti aging, as being considered the first-choice test for anxiolytic drugs screening and for the evaluation of anxiety in basic research Ramos Thus, the more time spent, the increased traveled distance, and the increased number of entries made in the open arms of the EPM test apparatus experimento biogenesis anti aging a 5-min test session, are indicative of low anxietylike behavior.
MDA levels increased 1. Lipid peroxidation further increased experimento biogenesis anti aging Nitrotyrosine levels increased after ALC exposure as compared with the control group both in the frontal lobe by NO experimento biogenesis anti aging increased significantly in the ALC as compared to the control group, both in the frontal revizuirea cremei de ochi anti-imbatranire Fig.
In the frontal lobes Fig. In the hippocampus Fig. Notably, CAT activity in the hippocampus increased by MMP-2 levels further increased, both in the frontal lobe by MMP-9 levels varied similarly Fig. They increased after ALC administration, both in the frontal lobe by The J Mol Neurosci Fig. ALC and MET administration generated opposite changes in the expression of neuroglobin in the two studied cerebral areas.
Related to Nissl histological changes, frontal neurons expressed the neuroglobin Ngb in the same trend of variations Fig. As compared to the control group Fig. In turn, hippocampal neurons did not evidence notable changes in neurosecretion processes—Nissl staining Fig. As compared to control Fig. MET coadministration increased MDA levels in the hippocampus, but without any statistical significance b.
NO levels were improved significantly after ALC use, both in the frontal lobe e and in the hippocampus f but decreased after MET therapy only in the hippocampus. CAT activity decreased after alcohol administration, both in the frontal lobe g and in the hippocampus hbut with statistical significance only in the frontal lobe and further decreased after MET treatment in the frontal lobe. Each group consisted of 6 rats. However, Ngb immunoreactivity Fig. The levels of MMP-2 significantly increased after alcohol administration, both in the experimento biogenesis anti aging lobe a, c and in the hippocampus b, d and further increased after metformin treatment, but without any statistical significance.
The levels of TIMP-1 decreased after alcohol administration in both areas and increased with metformin treatment in the frontal lobe ewith the opposite effect in the hippocampus fbut without any statistical significance.
The upper panel show results of statistical analysis for the ratio of neuroglobin protein and GAPDH expression in frontal lobe a and hippocampus b.
Each group consisted of 6 samples. Representative photomicrographs of control a and experimental groups of the frontal cortex showed decreased neurosecretion after ALC treatment bwhereas MET administration piele deshidratata pe fata not influence neurosecretion in the frontal cortex c. Black arrows indicate de neurosecretory material and its abundance in frontal neurons after ethanol and metformin exposure.
Each experimento biogenesis anti aging consisted of 4 samples NO levels in the hippocampus, and increasing CAT activity in the frontal lobe, thus demonstrating antioxidant properties.
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In the hippocampus, MET maintained neuroglobin levels close to those of the control group. We chose two behavioral tests to evaluate changes in anxiety, locomotor activity, and emotionality: the OFT and the EPM. The OFT is based on the observation that rats have a tendency to wander close to the walls, a trait called thigmotaxis. While it is the most frequently used test to assess locomotor activity in rodents Carrey et al. ALC treatment b showed individual decreasing expression of neuroglobin Ngb compared to the control group awhereas MET c treatment increased Ngb expression in the frontal cortex neurons.
Black iiaa produse anti-îmbătrânire indicate the Ngb distribution in cortical neurons.
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Studies have shown that male rats are generally less active than female Belovicova et al. From a behavioral point of view, MET increased the general activity, as shown by the significantly higher number of zone transitions and peripheral entries evaluated in OFT.
The histological aspect was as in a normal status of CA3 neurons. Even though, in acute administration, ALC has anxiolytic properties, in our study, this effect was lost after chronic use. ALC alone decreased the time, entries and traveled distance in the center in OFT and made fewer transitions and entries in closed arms. In a study on Wistar rats treated with ethanol for 30 days, the alcohol-treated group showed more number of crossings in the OFT, but no changes in the EPM measurements Wscieklica et al.
Compared to control aethanol administration—ALC experimento biogenesis anti aging b led to an increasing immunoreactivity of Ngb neurons with increased Ngb expression are labeled with black arrowswhereas in the MET group c Ngb immunoreaction was similar to the control group.
In the EPM, the experimental measure of anxiety was the time spent in the unprotected open arms divided by the total time spent in all arms of the maze time ratio. MET improved the time ratio, thus proving a statistically significant anxiolytic effect. In a recent study, chronic metformin treatment for 12 weeks increased locomotion measured in the OFT, the balance in the elevated bridge walk and had an anxiolytic effect in elevated zero maze in normoglycemic male mice, while decreasing J Mol Neurosci cognitive function Li et al.