miR-27b-3p Improved High Glucose-Induced Spermatogenic Cell Damage via Regulating Gfpt1/HBP Signaling

<b><i>Introduction:</i></b> Diabetes mellitus (DM)-induced testicular damage is characterized by abnormal apoptosis of spermatogenic cells. Here, we clarified the roles and the molecular mechanism of microRNA (miR)-27b-3p in high glucose (HG)-induced spermatogenic cell damage. <b><i>Methods:</i></b> GC-1 spg cells were treated with 30 mmol/L glucose for 24 h. Cell viability was assessed by 2.3 3-(4, 5-dimethylthiazolyl2)-2, 5-diphenyltetrazolium bromide (MTT) assay. And, levels of O-linked N-acetylglucosamine (OGT), apoptosis-related proteins, and autophagy-related proteins were evaluated using Western blot. Levels of tumor necrosis factor-α (TNF-α), IL-1β, IL-6, and UDP-N-acetylglucosamine (UDP-GlcNAc) were assessed by enzyme linked immunosorbent (ELISA) assay. Levels of reactive oxygen species (ROS), malonic dialdehyde (MDA) and activity of superoxide dismutase (SOD) in cells were determined using kits. Cell apoptosis was determined using flow cytometry assay. Besides, dual luciferase reporter assay was employed to verify the binding relationship between miR-27b-3p and glutamine-fructose-6-phosphate transaminase 1 (Gfpt1). <b><i>Results:</i></b> miR-27b-3p was markedly downregulated in HG-treated GC-1 spg cells. HG treatment caused decreased cell viability, increased oxidative stress and inflammation, and induced autophagy and apoptosis, which were abolished by miR-27b-3p overexpression. miR-27b-3p suppressed the activation of hexosamine biosynthetic pathway (HBP) signaling in HG-treated spermatogenic cells. miR-27b-3p directly bound to Gfpt1 and negatively regulated its expression. <b><i>Conclusion:</i></b> miR-27b-3p could improve HG-induced spermatogenic cell damage via regulating Gfpt1/HBP signaling, providing a new treatment strategy for the treatment of DM-induced testicular damage.

Biochemical characteristics of plant species of the genus Acer L.

The plants of the genus Acer section Palmata, which grow on the territory of M. M. Gryshko National Botanical Garden of NAS of Ukraine were investigated. It was found that the vast majority of A. palmatum and its cultivars were winter hardy. A study of the content of photosynthetic pigments revealed that the lowest content was in A. palmatum ‘Bloodgood’. The lowest content of malonic dialdehyde and proline was found in the leaves of A. palmatum ‘Dissectum’ and A. palmatum ‘Orange dream’, and the highest — in A. palmatum ‘Atropurpureum’ and A. palmatum. The greatest catalase activity was found in plants of A. palmatum ‘Atropurpureum’, and flavonoids — in A. palmatum.

Contemporary aspects of pain syndrome in endometriosis

The present investigation is devoted to the development of the pain syndrome in endometriosis patients. 150 women with a genital endometriosis of the reproductive age were investigated. On the base of the correlative analysis a high level of dependence between the pain syndrome severances, level of peroxidation (malonic dialdehyde) and level immune depression was revealed. Inflammation reaction (DTH) takes a significant role in the pain syndrome in endometriosis. Severance of the endometriosis development is not defined by the spreading stage, it is defined by the pain syndrome severance, which correlates with the anxiety level and depression and has extra-organic features.

Development of Inflammation in the Gastrointestinal Tract of Rats after Prolonged Administration of Alcohol

The purpose of the work was to experimentally study the chronic alcohol intoxication on the indicators of inflammation and lipid peroxidation in the gastrointestinal system. Materials and methods. Ethyl alcohol was added to the water for 2-month-old male rats, ranging from 5% to 15% for 108 days. In homogenates of mucous membranes of the gastrointestinal tract and liver, the activity of elastase enzymes, acid phosphatase and the concentration of malonic dialdehyde were determined, in serum – elastase activity and malonic dialdehyde content. Results and discussion. Biochemical research of one of the markers of inflammation (elastase activity) in rats found a probable increase of elastase activity in different parts of the digestive tract after prolonged alcohol consumption, regardless of the sex of the animals. Thus, in the serum of rats after the introduction of ethanol, the activity of elastase increased by 71.7%, in the oral mucosa – by 29.2%, in the gastric mucosa – by 55.5%, in the liver – by 29.0%. In the small and large intestine, the level of this marker of inflammation has changed slightly. The level of elastase activity shows the degree of accumulation of leukocytes in the tissues as a result of the development of the inflammatory process. Acid phosphatase activity in the oral mucosa of rats treated with ethanol increased by 47.4%, in the gastric mucosa – by 30.3%, in the mucous membrane of the small intestine – by 37.4%, in the mucous membrane of the colon – by 40.4%, in the liver – by 112.6%. Activation of acid phosphatase, along with other lysosomal enzymes, is the primary inflammatory response that triggers the production of mediators, which in turn cause secondary tissue alteration in subsequent stages of the inflammatory process. Therefore, the results obtained on the activation of acid phosphatase along with elastase indicate the presence of inflammation in the mucous membranes of the digestive tract, and especially in the liver of rats chronically treated with ethanol. The introduction of alcohol also led to an increase in the concentration of malonic dialdehyde in the mucous membranes: the oral cavity – by 20.3%, the stomach – by 32.3%, the small intestine – by 96.6%, the colon – by 50.2%, in the liver – by 39.4%, in serum – by 33.3%. A significant increase in the level of malonic dialdehyde in the tissues of the digestive tract of rats after long-term intake of ethanol is a sign of activation of lipid peroxidation and intensification of oxidative stress reactions. Conclusion. The results of the study of elastase activity indicate the development of inflammation in the mucous membranes of the gastrointestinal tract, liver and serum of rats under the influence of chronic administration of ethanol. Increased acid phosphatase activity in the tissues of the gastrointestinal tract after prolonged use of ethanol indicates damage to cell membranes, which is a consequence of inflammation. A significant increase in the level of malonic dialdehyde in the mucous membranes of the gastrointestinal tract, liver and serum of rats after chronic ethanol intake is a sign of intensification of oxidative stress reactions

miRNA-19b-3p Stimulates Cardiomyocyte Apoptosis Induced by Myocardial Ischemia Reperfusion via Downregulating PTEN

Objective. To clarify the function of miRNA-19b-3p in accelerating myocardial ischemia-reperfusion injury- (MIRI-) induced cardiomyocyte apoptosis by downregulating gene of phosphate and tension homology deleted on chromsome ten (PTEN), thus influencing the progression of acute myocardial infarction. Materials and Methods. miRNA-19b-3p and PTEN levels in HCM cells undergoing hypoxia/reoxygenation (H/R) were determined. Meanwhile, activities of myocardium injury markers [lactate dehydrogenase (LDH), malondialdehyde; malonic dialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX)] in H/R-induced HCM cells were tested. Through dual-luciferase reporter gene assay, the binding between miRNA-19b-3p and PTEN was verified. Regulatory effects of miRNA-19b-3p and PTEN on apoptotic rate and apoptosis-associated gene expressions (proapoptotic protein Bcl-2 associated X protein (Bax), antiapoptotic protein B-cell lymphoma-2 (Bcl-2), and cytochrome C) in H/R-induced human cardiac myocytes (HCM) cells were examined. Results. miRNA-19b-3p was upregulated, while PTEN was downregulated in H/R-induced HCM cells. Knockdown of miRNA-19b-3p decreased activities of LDH, MDA, and GSH-PX, but increased SOD level in H/R-induced HCM cells. The binding between miRNA-19b-3p and PTEN was confirmed. More importantly, knockdown of miRNA-19b-3p reduced apoptotic rate, downregulated proapoptosis gene expressions (Bax and cytochrome C), and upregulated antiapoptosis gene expression (Bcl-2), which were reversed by silence of PTEN. Conclusions. miRNA-19b-3p is upregulated in HCM cells undergoing hypoxia and reoxygenation, which accelerates MIRI-induced cardiomyocyte apoptosis through downregulating PTEN.

Male Infertility: Pathogenetic Significance of Oxidative Stress and Antioxidant Defence (Review)

The basis of the pathogenesis of male infertility is the processes of peroxide oxidation of biological substrates, especially lipids and proteins. By destroying the sperm membrane, toxic peroxidation products reduce its motility and ability to fertilize the egg, which is determined by a decrease in the number of motile sperm in the ejaculate. These changes lead to complete or partial male infertility. The authors of the review found that is accompanied by a damaging effect on the structural and functional activity of the gonads and is manifested, in particular, by an imbalance in the hormonal background of the male body. Similar effects are characteristic of an increase in the content of reactive Nitrogen species and its metabolites, which cause nitrosative stress, which is also the cause of male hypofertility and is inseparable from the state of oxidative stress. In scientific work it is determined that the accumulation of harmful peroxidation products leads to damage and destruction of sperm DNA, reduced activity of acrosomal enzymes and mitochondrial potential of sperm, reduced overall antioxidant activity. This makes it impossible for an adequate response of the body. Multi component antioxidant defense system resists stress. It is represented by enzymatic and non-enzymatic links, which can neutralize harmful radicals and peroxidation products. It contributes to the full manifestation of reproductive function. The presence of powerful antioxidant properties of catalase, superoxide dismutase, and enzymes of the thiol-disulfide system, which form the enzymatic system of antioxidant protection, as well as selenium, zinc, copper, other trace elements, retinol, tocopherol, ascorbic acid, and vitamins as parts of the non-enzymatic system is shown. The efficiency of registration is substantiated thin biochemical shift detectors or complex methods, such as total antioxidant status of sperm or sperm plasma, mitochondrial membrane potential, etc along with simple markers of oxidative stress, such as diene conjugates, malonic dialdehyde, and metabolites of the Nitrogen Oxide cycle. Given the leading role of oxidative stress in the development of male hypofertility, the prospect of further research is the search for modern means for correction, especially among substances with pronounced redox activity

The systemic pro- and antioxidant processes in preschool children with recurrent acute respiratory diseases

Background. The imbalance of innate defense early mechanisms in children from pathogens — peroxidation and antioxidant system, can lead to a decrease in the efficiency of the entire immune system. The study of the lipid peroxidation (LPO) and antioxidant protection (AOP) status in children with recurrent acute respiratory diseases will complement the scientific data on the antimicrobial immunity formation processes. The study was aimed to increase the information value of timely detected pro- and antioxidant processes disorders in children with recurrent acute respiratory diseases by studying the primary, secondary, tertiary, and quaternary compounds of LPO and AOP in the serum. Materials and methods. Sixty children aged from 2 to 5 years were examined. Two groups were formed: group 1 — children with respiratory acute infectious more than six times per year (n = 30); group 2 — children with respiratory acute infectious six or fewer times per year (n = 30). Results. The children in group 1 more often presented with high serum levels of lipid hydroperoxides (by 80.0 %; p < 0.05), diene conjugates (by 80.0 %; p < 0.05), malonic dialdehyde (76.7 %; p < 0.05), diene ketones (76.7 %; p < 0.05), Schiff bases (76.7 %; p < 0.05). ceruloplasmin (80.0 %; p < 0.05), superoxide dismutase (80.0 %; p < 0.05), glutathione peroxidase (86.7 %; p < 0.05), catalase activity (86.7 %; p < 0.05). The values of retinol, tocopherol, ascorbic acid in children in the observation groups did not differ statistically significantly. Conclusions. The children aged from 2 to 5 years old with recurrent acute respiratory diseases have an imbalance of the LPO and AOP systems. It was characterized by a simultaneous increase in the serum content of the primary, secondary, and end-products of LPO, enzymes of anti-peroxide and anti-oxygen protection against the background of the lack of adaptive increase in the content of anti-radical compounds.

CHANGES IN ACTIVITY OF INDUCIBLE NITRIC OXIDE SYNTHASE IN EXPERIMENTAL DIABETIC RETINOPATHY AND METHODS FOR ITS CORRECTION

Diabetic retinopathy according to the World Health Organization reports is known as the main cause of decreased vision and blindness in diabetes. The purpose of the study is to analysis of changes in the activity of inducible nitric oxide synthase in experimental diabetic retinopathy and in various methods of its correction. Materials and methods. The research was carried out on white Wistar rats weighting 180-220 g. The animals were divided into 7 groups: 1st group included 60 intact animals; 2nd group involved 60 animals with modelled diabetic retinopathy (DR) without further correction; 3rd group included 60 animals with modelled DR and subsequent hyperglycemia correction; 4th group included 60 animals with modelled DR, which received subsequent hyperglycemia correction with aflibercept and L-arginine solution administration; 5th group consisted of 60 animals with modelled DR and subsequent hyperglycemia correction by aflibercept and bromfenac administration; 6th group was formed by 60 animals with modelled DR and subsequent hyperglycemia correction by aflibercept, L-carnitine and bromfenac administration; and the 7th group included 60 animals with modelled DR and subsequent hyperglycemia correction by aflibercept, L-arginine solution and citicoline. Results and conclusion. The results obtained demonstrate an increase in the activity of inducible nitric oxide synthase, starting from the 30th day of the experiment and with subsequent progression to the 60 and 108 days of experimental diabetic retinopathy that points out the deterioration of the physiological pathway of nitric oxide synthesis. Correction with hypoglycemic agents in group 3 had a positive effect, but was not able to reduce the activity of inducible nitric oxide synthase, which increased in the 2nd and 3rd stages, thus, there was a need for additional agents. The use of aflibercept and nitric oxide donor in group 4 to correct the progression of diabetic retinopathy significantly reduced the activity of inducible nitric oxide synthase (most effectively on the one hundred and eighty days of the experiment), but did not reach the control values. It has been proved that the correction applied in groups 5 and 6 significantly reduced the activity of inducible nitric oxide synthase but did not reach the control values. Moreover, the marker activity in the group 5 grew up on the 108 day. The study has shown the most effective correction includes a combination of metformin, aflibercept, L-arginine and citicoline given to the rats of group 7, as evidenced by the normalization of malonic dialdehyde levels on the 60 day of the experiment; on the 108 days of experiment there has been a decrease in marker content to control values.

Metabolic Changes in Seed Embryos of Hypoxia-Tolerant Rice and Hypoxia-Sensitive Barley at the Onset of Germination

Rice (Oryza sativa L.) and barley (Hordeum vulgare L.) are the cereal species differing in tolerance to oxygen deficiency. To understand metabolic differences determining the sensitivity to low oxygen, we germinated rice and barley seeds and studied changes in the levels of reactive oxygen species (ROS) and reactive nitrogen species (RNS), activities of the enzymes involved in their scavenging, and measured cell damage parameters. The results show that alcohol dehydrogenase activity was higher in rice than in barley embryos providing efficient anaerobic fermentation. Nitric oxide (NO) levels were also higher in rice embryos indicating higher NO turnover. Both fermentation and NO turnover can explain higher ATP/ADP ratio values in rice embryos as compared to barley. Rice embryos were characterized by higher activity of S-nitrosoglutathione reductase than in barley and a higher level of free thiols in proteins. The activities of antioxidant enzymes (superoxide dismutase, ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase) in imbibed embryos were higher in rice than in barley, which corresponded to the reduced levels of ROS, malonic dialdehyde and electrolyte leakage. The observed differences in metabolic changes in embryos of the two cereal species differing in tolerance to hypoxia can partly explain the adaptation of rice to low oxygen environments.

Immunological and genetic indices in workers under long-term exposure to low-doses of acrylonitrile

Introduction. Nowadays there is very relevant research on the study of the characteristics of the impact on the health of workers of low levels of harmful factors (acrylonitrile) of production during long-term exposure. Aim of the study was to examine peculiarities of immunologic and genetic indices in workers under the long-term exposure to acrylonitrile in low doses. Materials and methods. Our research object was working area air (MPCw.ar.=0.5 mg/m3) and biological media (blood and exhaled air) of workers employed at industrial rubber manufacture. Acrylonitrile was determined via a non-invasive procedure in exhaled air with samples being concentrated on sorption tubes that were then analyzed with capillary gas chromatography. Blood samples were examined to determine contents of malonic dialdehyde, lymphocytes (absolute and relative activated T-lymphocytes CD3+CD25+, absolute and relative activated T-lymphocytes CD3+CD95+), cytokines (VEGF), oncomarkers (PSA), and adrenals hormones; to do that, we applied ELISA tests and flow cytometry. Results. Acrylonitrile was established to occur in working area air in concentrations varying within MPCw.ar. range (0.007-0.015 mg/m3) being 2-3 times higher than in air inside offices at the same enterprise. We obtained statistically significant linear dependence between concentrations of acrylonitrile in the air exhaled by workers (y) and their working experience (x) that was given with the following equation: y=0.00046+0.00027x. According to the results of the laboratory examination of the workers, violations of the antioxidant defense were established. Contents of malonic dialdehyde and steroid hormones including progesterone, estradiol, and hydrocortisone that were pathogenetically linked to each other were authentically up to 3.2 times higher in the test group than in the reference one (p<0.05). Risk for antioxidant protection disorders such as elevated malonic dialdehyde contents in blood plasma might occur in the test group was 1.58 times higher than in the reference one. Conclusion. We revealed certain peculiarities in polymorphism of PPARGC1A Gly482Ser rs8192678 gene, the variability of which contributes to the formation of pathology of the cardiovascular, endocrine systems, oncoproliferative states that increase the likelihood of these undesirable events.