Neuroprotective Potential of Tannic Acid Against Neurotoxic Outputs of Monosodium Glutamate in Rat Cerebral Cortex.

Glutamate in monosodium glutamate (MSG), which is widely used in the food industry, has an important role in major brain functions such as memory, learning, synapse formation, and stabilization. However, extensive use of MSG has been linked with neurotoxicity. Therefore, in addition to clarifying the underlying mechanisms of MSG-induced neurotoxicity, it is also important to determine safe agents that can diminish the damage caused by MSG. Tannic acid (TA) is a naturally occurring plant polyphenol that exhibits versatile physiological effects such as anti-inflammatory, anti-carcinogenic, antioxidant, and radical scavenging. This study was conducted to assess the neurotoxic and neuroprotective effects of these two dietary components in the rat cerebral cortex. Twenty-four Sprague Dawley rats were divided into 4 equal groups and were treated with MSG (2 g/kg) and TA (50 mg/kg) alone and in combination for 3 weeks. Alterations in oxidative stress indicators (MDA and GSH) were measured in the cortex tissues. In addition, changes in enzymatic activities and gene expression patterns of antioxidant system components (GST, GPx, CAT, and SOD) were investigated. Furthermore, mRNA expressions of FoxO transcription factors (Foxo1 and Foxo3) and apoptotic markers (Casp3 and Casp9) were assessed. Results revealed that dietary TA intake significantly rehabilitated MSG-induced dysregulation in cortical tissue by regulating redox balance, cellular homeostasis, and apoptosis. The present study proposes that MSG-induced detrimental effects on cortical tissue are potentially mitigated by TA via modulation of oxidative stress, cell metabolism, and programmed cell death.

Tannic acid may ameliorate doxorubicin-induced changes in oxidative stress parameters in rat spleen.

Doxorubicin (DOX) is a potent and broad-spectrum drug widely used in the treatment of cancer. However, the toxicity and side effects of DOX on various organs limit its clinical use. Approaches using natural antioxidants with these drugs have the potential to alleviate negative side effects. The aim of this study was to investigate the potential protective effect of tannic acid, a polyphenolic compound found naturally in plants, against DOX-induced spleen toxicity. Expression levels of Alox5, Inos, IL-6, Tnf-α, Casp-3, Bax, SOD, GST, CAT and GPx genes were determined using cDNAs obtained from spleen tissues of rats treated with DOX, tannic acid and both. In addition, SOD, CAT, GPx and GST enzyme activities, and GSH and MDA levels were measured in tissues. In the spleen tissues, DOX caused a decrease in the level of GSH and an increase in the level of MDA. In addition, it was determined that DOX had a suppressive effect on CAT, GST, SOD and GPx mRNA levels and its enzyme activities, which are antioxidant system components. The mRNA expression levels of proinflammatory cytokine markers, apoptotic genes, and some factors involved in cell metabolism showed a change compared to the control after DOX application. However, as a result of tannic acid treatment with DOX, these changes approached the values of the control group. The findings showed that tannic acid had a protective effect on the changes in the oxidative stress and inflammation system in the rat spleen as a result of the application of tannic acid together with DOX.

Investigation of the multi-targeted protection potential of tannic acid against doxorubicin-induced kidney damage in rats.

Doxorubicin (DOX) is an antitumor drug that is powerful but can cause worse outcomes, including nephrotoxicity, and therefore has limited clinical use. Therefore, it is necessary to identify safer agents that can minimize the damage caused by the drug without shifting the treatment performance, in addition to clarifying the underlying mechanisms of DOX-induced aberrant in vivo renal activation. In this study, we tested the prophylactic capacity and mechanisms of action of tannic acid (TA) against DOX-mediated kidney damage in rats and evaluated the nephrotoxic activity of DOX when used with TA. Rats were treated during the two weeks with cumulative (18 mg/kg with six different injections) DOX, daily TA (50 mg/kg), and the DOX + TA combination. Changes in major metabolites and components involved in antioxidant metabolism were evaluated in the kidney tissues of all animals. Further, the gene expression levels of regulatory factors that have critical importance in cell metabolism, inflammation, and apoptosis were investigated. Both biochemical and molecular examinations showed that TA improved DOX-induced dysregulations at both protein and gene levels in the kidneys. Increased lipid peroxidation and decreased glutathione levels were reversed. Consistent with oxidative stress marker metabolites, suppressed antioxidant enzyme activities and transcript levels of antioxidant system members were restored. Of note, combination treatment with TA could overcome doxorubicin-induced gene expressions markedly altered by DOX, suggesting that nephroprotection conferred by TA involved the remodeling of stress resistance, cell metabolism, inflammation, and apoptosis. Collectively, the present in vivo study suggests that TA could be used as a multitarget and effective agent for the mitigation of doxorubicin-induced nephrotoxicity without changing the therapeutic efficacy of the drug.

Integrated Bioinformatics Analysis to Identify Alternative Therapeutic Targets for Alzheimer's Disease: Insights from a Synaptic Machinery Perspective.

Alzheimer's disease (AD), the most common type of dementia, is a serious neurodegenerative disease that has no cure yet, but whose symptoms can be alleviated with available medications. Therefore, early and accurate diagnosis of the disease and elucidation of the molecular mechanisms involved in the progression of pathogenesis are critically important. This study aimed to identify dysregulated miRNAs and their target mRNAs through the integrated analysis of miRNA and mRNA expression profiling in AD patients versus unaffected controls. Expression profiles in postmortem brain samples from AD patients and healthy individuals were extracted from the Gene Expression Omnibus database and were analyzed using bioinformatics approaches to identify gene ontologies, pathways, and networks. Finally, the module analysis of the PPI network and hub gene selection was carried out. A total of five differentially expressed miRNAs were extracted from the miRNA dataset, and 4312 differentially expressed mRNAs were obtained from the mRNA dataset. By comparing the DEGs and the putative targets of the altered miRNAs, 116 (3 upregulated and 113 downregulated) coordinated genes were determined. Also, six hub genes (SNAP25, GRIN2A, GRIN2B, DLG2, ATP2B2, and SCN2A) were identified by constructing a PPI network. The results of the present study provide insight into mechanisms such as synaptic machinery and neuronal communication underlying AD pathogenesis, specifically concerning miRNAs.

Molecular docking and inhibition studies of vulpinic, carnosic and usnic acids on polyol pathway enzymes.

Aldose reductase (AR) and sorbitol dehydrogenase (SDH) are important enzymes of the polyol pathway. In the current study, inhibitory effects of vulpinic acid (VA) carnosic acid (CA) and usnic acid (UA) on purified AR and SDH enzymes were determined. These enzymes inhibition could be essential to prevent diabetic complications. AR and SDH enzymes were purified from sheep kidney. Then, VA, CA and UA were tested in various concentrations against these enzymes activity in vitro. KI values were found to be as 1.46 ± 0.04, 5.13 ± 0.25 and 11.71 ± 0.27 µΜ for VA, CA and UA, respectively, for AR. KI constants were found to be as 15.32 ± 0.34, 145.60 ± 2.17 and 213.40 ± 2.64 µΜ VA, CA and UA, respectively, for SDH. These findings indicate that VA, CA and UA could be useful in the treatment of diabetic complications.Communicated by Ramaswamy H. Sarma.

A bioinformatics approach for identifying potential molecular mechanisms and key genes involved in COVID-19 associated cardiac remodeling.

In 2019 coronavirus disease (COVID-19), whose main complication is respiratory involvement, different organs may also be affected in severe cases. However, COVID-19 associated cardiovascular manifestations are limited at present. The main purpose of this study was to identify potential candidate genes involved in COVID-19-associated heart damage by bioinformatics analysis. Differently expressed genes (DEGs) were identified using transcriptome profiles (GSE150392 and GSE4172) downloaded from the GEO database. After gene and pathway enrichment analyses, PPI network visualization, module analyses, and hub gene extraction were performed using Cytoscape software. A total of 228 (136 up and 92 downregulated) overlapping DEGs were identified at these two microarray datasets. Finally, the top hub genes (FGF2, JUN, TLR4, and VEGFA) were screened out as the critical genes among the DEGs from the PPI network. Identification of critical genes and mechanisms in any disease can lead us to better diagnosis and targeted therapy. Our findings identified core genes shared by inflammatory cardiomyopathy and SARS-CoV-2. The findings of the current study support the idea that these key genes can be used in understanding and managing the long-term cardiovascular effects of COVID-19.

Identification of hub genes associated with obesity-induced hepatocellular carcinoma risk based on integrated bioinformatics analysis.

Obesity, which has become one of the biggest public health problems of the twenty-first century, accompanies many chronic conditions, including cancer. On the other hand, liver cancer, which is known to be associated with obesity, is considered another serious threat to public health. However, the underlying drivers of the development of obesity-associated hepatocellular carcinoma (HCC) remain blurry. The current study attempted to identify the key genes and pathways in the obesity-induced development of HCC using integrated bioinformatics analyses. Obesity and HCC-associated gene expression datasets were downloaded from Gene Expression Omnibus (GEO) and analyzed to identify overlapping differentially expressed genes (DEGs) and hub genes. The prognostic potentials, survival analysis, and expression levels of hub genes were further assessed. Moreover, the correlation between hub genes and the immune cells infiltration was analyzed. The findings of this research revealed that both mRNA and protein expression levels of the four hub genes (IGF1, ACADL, CYP2C9, and G6PD) involved in many important metabolic pathways are remarkably altered in both obese individuals and patients with HCC. The results demonstrated that these dysregulated genes in both obesity and HCC may serve as considerable targets for the prevention and treatment of HCC development in obese individuals.

Muscle cellularity, growth performance and growth-related gene expression of juvenile climbing perch Anabas testudineus in response to different eggs incubation temperature.

Although indigenous climbing perch (Anabas testudineusis) is a highly valuable species, slow growth pattern during the culture period impeding its commercial success in aquaculture. In many fish species, it has been demonstrated that incubation temperature of eggs influenced the muscle development and growth rates, which persisted throughout the subsequent larval and juvenile phases. Therefore, this study aimed to investigate whether different incubation temperature of eggs prior to hatching can stimulate the muscle development, growth, and growth-related gene expression of the slow-growing indigenous species of climbing perch. The fertilized eggs of A. testudineus from an artificial breeding program were incubated under control temperature of 24 °C (IT24), 26 °C (IT26), 28 °C (IT28), and 30 °C (IT30) in 10L glass aquaria with four replicated units for each temperature treatment. After hatching, the larvae from each incubated temperature were separately reared at ambient temperature for 10 days in aquarium, 20 days in hapas, and the next 42 days in cages, totaling 72 days post-hatching (dph). The hatching rates were found significantly (P < 0.05) higher in IT28 compared to the other incubation temperature treatments. After 72 dph, the growth performances (%length gained, %weight gained and SGR) were found to be significantly highest (P < 0.05) in the IT28, followed by the treatments IT30, IT26, and IT24, respectively. Survival rate (73 ± 1.257%) was also found to be highest in the same treatment. The rate of new muscle fiber formation was identified to be significantly highest (P < 0.05) in IT28 followed by the IT26, IT30 and IT24, respectively. The relative mRNA expression level of GHRH, IGF1, IGF2 and PRL was also significantly highest in the IT28 (P < 0.05) compared to other treatments. Results from the present study clearly suggested that 28 °C is the optimum eggs incubation temperature of the native strain of A. testudineus for its highest growth performances in captive breeding condition.

Effect of a Prolonged Dietary Iron Intake on the Gene Expression and Activity of the Testicular Antioxidant Defense System in Rats.

Despite the fact that iron represents a crucial element for the catalysis of many metabolic reactions, its accumulation in the cell leads to the production of reactive oxygen species (ROS), provoking pathological conditions such as cancer, cardiovascular diseases, diabetes, neurodegenerative diseases, and fertility. Thus, ROS are neutralized by the enzymatic antioxidant system for the purpose of protecting cells against any damage. Iron is a potential risk factor for male fertility. However, the mechanism of action of iron on the testicular antioxidant system at the gene and protein levels is not fully understood. Thus, the purpose of the current research was to ensure a better understanding of how the long-term iron treatment influences both gene expression and enzyme activities of the testicular antioxidant system in rat testis. The data of our study showed that a significant dose-dependent increase occurred in the iron level in rat testis. A reduction occurred in reduced glutathione (GSH) levels, which represent a marker of oxidative stress, along with long-term iron overload. The expression and activity of glucose 6-phosphate dehydrogenase (G6pd), glutathione reductase (Gr), glutathione peroxidase (Gpx), and glutathione S-transferases (Gst) were significantly affected by the presence of iron. The findings of the current research demonstrate that the long-term toxic dietary iron overload influences the gene expression and enzyme activity of the testicular antioxidant defense system, but the actual effect occurs at the protein level. This may modify the sperm function and dysfunction of the male reproductive system.

Examining the link between dose-dependent dietary iron intake and Alzheimer's disease through oxidative stress in the rat cortex.

BACKGROUND: Neurodegenerative diseases such as Alzheimer's and Parkinson's disease are characterized by the progressive deterioration of the structure and function of the nervous system. A number of environmental risk factors including potentially toxic elements such as iron, lead to negative effects on many metabolic reactions as well as neuroprotection. The aim of this study is to reveal whether long-term iron overload is one of the underlying factors in the pathogenesis of Alzheimer's disease (AD). METHODS: 15 young-adult male rats were randomly divided into 5 groups treated with iron through drinking water for 4 months. Following feeding, the iron content, reduced glutathione (GSH), and hydrogen peroxide (H2O2) levels of cortex tissues were measured. Specific enzyme activities were determined spectrophotometrically. mRNA expression profiles were measured using real-time PCR (qPCR). RESULTS: Iron levels were elevated in case of non-toxic (0.87 and 3 µg/mL) iron administration. However, no changes were observed in toxic (30 and 300 µg/mL) iron administration. GSH and H2O2 levels altered with long-term iron overload. Glutathione peroxidase (GPx) enzyme activities significantly increased in all groups, while glutathione S-transferase (GST) activity increased only in case of 0.87 and 30 µg/mL iron administration. Expression levels of neuroprotective and AD-related genes were altered by 3 µg/mL iron overload in a dose-dependent manner. The expression and activity of acetylcholinesterase (AChE) were elevated at 3 µg/mL iron concentration. CONCLUSION: The findings of the present study allow us to conclude that long-term dietary iron intake, especially at a dose of 3 µg/mL demonstrates negative effects on the rat cortex by provoking antioxidant metabolism and AD pathology in a dose-dependently.

Inhibitory Effects of Usnic and Carnosic Acid on Some Metabolic Enzymes: An In vitro Study.

BACKGROUND: Natural products are produced via primary and secondary metabolism in different organisms. The compounds obtained via secondary metabolism are not essential for the survival of the organism, but they can have a different value for humans. OBJECTIVE: The objective of this study was to examine inhibitory effects of Usnic Acid (UA), a well-known lichen secondary metabolite, and Carnosic Acid (CA), the primary antioxidant compound of Rosmarinus officinalis L., on purified Human Paraoxonase, (PON1), Glutathione Reductase (GR) and Glutathione S-Transferase (GST). These enzymes have antioxidant properties and a protective effect on the oxidation of free radicals. Hence, deficiencies of such enzymes inside cells can result in a buildup of toxic substances and cause some metabolic disorders. METHODS: UA and CA were tested in various concentrations against human GST, PON1, and GR activity in vitro and they reduced human GST, PON1, and GR activity. RESULTS: UA Ki constants were calculated as 0.012±0.0019, 0.107±0.06 and 0.21±0.1 mM for GST, PON1, and GR enzymes. CA Ki constants were determined as 0.028±0.009, 0.094±0.03 and 0.79±0.33 mM, for GST, PON1, and GR enzymes. UA and CA showed competitive inhibition for GR and GST enzymes, while they exhibited non-competitive inhibition for PON1. CONCLUSION: These findings indicate that UA and CA could be useful in drug development studies.

Multiple transcription factors mediating the expressional regulation of myosin heavy chain gene involved in the indeterminate muscle growth of fish.

Torafugu myosin heavy chain gene, MYHM2528-1, is specifically expressed in neonatal slow and fast muscle fibers, suggesting its functional role in indeterminate muscle growth in fish. However, the transcriptional regulatory mechanisms of MYHM2528-1 involved in indeterminate muscle growth in fish remained unknown. We previously isolated a 2100 bp 5'- flanking sequence of torafugu MYHM2528-1 that showed sufficient promoter activity to allow specific gene expression in neonatal muscle fibers of zebrafish. Here, we examined the cis-regulatory mechanism of 2100 bp 5'-flanking region of torafugu MYHM2528-1 using deletion-mutation analysis in zebrafish embryo. We discovered that myoblast determining factor (MyoD) binding elements play a key role and participate in the transcriptional regulation of MYHM2528-1 expression in zebrafish embryos. We further discovered that paired box protein (Pax3) are required for promoting MYHM2528-1 expression and myocyte enhancer factor-2 (MEF2) binding sites participate in the transcriptional regulation of MYHM2528-1 expression in slow/fast skeletal muscles. Our study also confirmed that the nuclear factor of activated T-cell (NFAT) binding sites take part in the transcriptional regulation of MYHM2528-1 expression in slow and fast muscles fiber in relation to indeterminate muscle growth. These results obviously confirmed that multiple cis-elements in the 5'-flanking region of MYHM2528-1 function in the transcriptional regulation of its expression.

In vitro and in vivo effects of iron on the expression and activity of glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and glutathione reductase in rat spleen.

Iron is an indispensable element for vital activities in almost all living organisms. It is also a cofactor for many proteins, enzymes, and other essential complex biochemical processes. Therefore, iron trafficking is firmly regulated by Hepcidin (Hamp), which is regarded as the marker for iron accumulation. The disruption of iron homeostasis leads to oxidative stress that causes various human diseases, but this mechanism is still unclear. The aim of this study is to provide a better in vivo and in vitro understanding of how long-term iron overload affects the gene expression and activities of some antioxidant enzymes, such as glucose 6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), and glutathione reductase (GR) in the spleen. The findings of this study show that iron overload reduces the gene expression of G6pd, 6pgd, and Gr, but its actual effect was on the protein level.

An approach to clarify the effect mechanism of glyphosate on body malformations during embryonic development of zebrafish (Daino rerio).

In this study, it has been investigated that the effects of glyphosate, which is a herbicide within organophosphate and unselective widely used in agriculture on enzyme activity of carbonic anhydrase, production of reactive oxygen species, cell apoptosis and body morphology during the embryonic development of zebrafish. To this end, it has been treated embryo with 1, 5, 10 and 100 mg/L gyphosate at 96 h. The embryos treated with glyphosate from 4 hpf were evaluated by considering the survival rates, hatching rates, body malformations under the stereo microscope in 24, 48, 72 and 96th hours. In order to clarify the mechanism of the abnormalities ROS, enzyme activity of carbonic anhydrase and cellular death were detected end of the 96th hour. The data obtained in the present study have shown that glyphosate treatment inhibited CA activity, caused production of ROS especially branchial regions, triggered cellular apoptosis and caused several types of malformations including pericardial edema, yolk sac edema, spinal curvature and body malformation in a dose-dependent manner. As a conclusion, in light of present and previous studies, we can deduce that (1) the probable reason of ROS production was CA inhibition via decreasing of CO2 extraction and developing respiratory acidosis (however, one needs to clarify), (2) abundance of ROS triggered cellular apoptosis and (3) as a result of cellular apoptosis malformations increased. These data will enable us to further understand potential toxic mechanism of glyphosate on embryonic development stage of zebrafish and may be useful for assessment in the toxicology studies.

Can chondral healing be improved following microfracture? The effect of adipocyte tissue derived stem cell therapy.

BACKGROUND: We aimed to investigate the effect of adipose tissue-derived mesenchymal stem cells (ADSCs) on chondral healing using the microfracture (MF) technique. METHODS: Thirty male rabbits were randomly divided into three groups. Standard cylindrical osteochondral defects (OCDs) were created in the weight-bearing areas of the medial condyles of all the right knees; the defects were four millimeters in diameter and two millimeters in depth. The control group (group A) was restricted to spontaneous healing. For group B, we performed MF with a 1.5-mm drill. For group C, we applied MF using the same method and then applied 3×10(6) ADSCs to the defect area. At eight weeks post-operation, the subjects were sacrificed, and the distal femoral joint surfaces were evaluated histopathologically for chondral healing. The samples were scored according to the International Cartilage Repair Society (ICRS) scale. RESULTS: The results for group C were significantly better than those for group A in terms of the surface properties (p=0.003). The matrix evaluation was better for group A than for group C (p=0.01). The cell distribution, cell viability and subchondral bone parameters were similar between the groups (p=0.198, p=0.387 and p=0.699). The cartilage mineralization parameter was better for group C than for group A (p=0.001). The signs of healing were better for group C than for group B, but the differences were not significant (p=0.185). CONCLUSIONS: Improvements with additional ADSC treatments were not statistically significant in cases in which ADSC treatment was compared with isolated MF treatment. CLINICAL RELEVANCE: Additional ADSCs treatment may have positive effect on chondral healing but it doesn't seem significant.

Stimulation of gene expression and activity of antioxidant related enzyme in Sprague Dawley rat kidney induced by long-term iron toxicity.

The trace elements such as iron are vital for various enzyme activities and for other cellular proteins, but iron toxicity causes the production of reactive oxygen species (ROS) that causes alterations in morphology and function of the nephron. The present study was designed to determine the effect of long-term iron overload on the renal antioxidant system and to determine any possible correlation between enzymatic and molecular levels. Our data showed that reduced glutathione (GSH) levels, which is a marker for oxidative stress, strikingly decreased with a long-term iron overload in rat kidney. While renal mRNA levels of glucose 6-phosphate dehydrogenase (G6pd), 6-phosphogluconate dehydrogenase (6pgd) and glutathione peroxidase (Gpx) were significantly affected in the presence of ferric iron, no changes were seen for glutathione reductase (Gsr) and glutathione S-transferases (Gst). While the iron affected the enzymatic activity of G6PD, GSR, GST, and GPX, it had no significant effect on 6PGD activity in the rat kidney. In conclusion, we reported here that the gene expression of G6pd, 6pgd, Gsr, Gpx, and Gst did not correlate to enzyme activity, and the actual effect of long-term iron overload on renal antioxidant system is observed at protein level. Furthermore, the influence of iron on the renal antioxidant system is different from its effect on the hepatic antioxidant system.

Expression of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in oxidative stress induced by long-term iron toxicity in rat liver.

Reactive oxygen species (ROS) are highly reactive and oxygen-containing molecules that are derived by metabolic activities or from environmental sources. Toxicity of heavy metals including iron has the ability to generate ROS in all living organisms. The pentose phosphate pathway enzymes, which are glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase, produce nicotinamide adenine dinucleotide phosphate (NADPH) that enables cells to counterbalance the oxidative stress via the action of the glutathione system. The results presented here have shown that toxic and nontoxic levels of iron have a strong effect on the expression of both genes. While toxic levels of iron exhibited significant changes in enzyme activity, nontoxic levels had no effect on enzymes in rat liver. Our results are the first evidence to elucidate how oxidative stress induced by long-term iron toxicity affects both enzymes at the enzymatic and molecular level and also to determine any possible correlation between the enzymatic and molecular levels.

Impact of long term Fe³âº toxicity on expression of glutathione system in rat liver.

The free radicals within the body, produced by metabolic activities or derived from environmental sources are relatively related to hepatoxicity. Since heavy metals including iron have the ability to produce free radicals, the liver glutathione system neutralizes them to protect cells against any damage. The objective of this study is to indicate the toxic effects of iron on the glutathione system at the enzymatic and molecular level. Thus, any possible correlation between enzymatic and molecular levels can be determined. According to our results, while mRNA expression of glutathione reductase (Gsr) and glutathione S-transferases (Gsta5) genes were not affected by long-term exposure to various concentrations of iron (Fe(3+)), transcription level of glutathione peroxidase (Gpx2) was influenced in the presence of toxic iron. Whereas the enzyme activites of GSR (GR), GPX and GST were significantly affected in rat liver.