iNOS Activity Research Articles

ITreg cells-secreted IL10 alleviates lupus nephritis through inactivating lncRNA HAR1A transcription to suppress SMARCD1-mediated iNOS activation

Lupus nephritis (LN) is a highly prevalent complication of systemic lupus erythematosus (SLE). Long non-coding RNAs (lncRNAs) are essential modulators in multiple types of human diseases, including LN. In the current study, we searched on online GEO database to select out lncRNAs that were differentially expressed in blood samples of LN patients. Through further RT-qPCR analysis, we found that lncRNA Highly Accelerated Region 1 A (HAR1A) is most significantly upregulated in blood samples of LN patients. Functionally, we detected that overexpression of HAR1A could aggravate LPS-induced injury and inflammation. According to the results of bioinformatics analysis and mechanism experiments, we determined that HAR1A binds with miR-149-3p to upregulate SMARCD1 through competing endogenous RNA (ceRNA) mechanism. It has been proven that iNOS is an inflammation inducer. Here, we found that HAR1A/miR-149-3p/SMARCD1 upregulates iNOS expression through enhancing H3K27ac level in iNOS promoter. Previously, we proved that CD8+CD103+ iTreg cells could alleviate glomerular endothelial cell injury. Moreover, we demonstrated that CD8 + CD103+ iTreg cells-secreted IL-10 downregulated HAR1A expression by impeding NF-κB pathway activation. In conclusion, this study provides evidences revealing a novel molecular pathway blocked by CD8+CD103+ iTreg cells in LN.

Normalization of CF Immune System Reverses Intestinal Neutrophilic Inflammation and Significantly Improves the Survival of CF Mice

Cystic fibrosis (CF) is an autosomal recessive genetic disorder, affecting multiple organ systems. CF intestinal disease develops early, manifesting as intestinal bacterial overgrowth/dysbiosis, neutrophilic inflammation and obstruction. As unresolvable infection and inflammation reflect host immune deficiency, we sought to determine if the CF-affected immune system plays any significant role in CF intestinal disease pathogenesis. CF and sibling wild-type (WT) mice underwent reciprocal bone marrow transplantation. After immune reconstitution, their mortality, intestinal transit, fecal inflammatory markers, and mucosal immune cell composition were assessed. Moreover, reciprocal neutrophil transfusion was conducted to determine if neutrophil function affects intestinal movement. Furthermore, expression of induced nitric oxide synthase (iNOS) and production of nitric oxide (NO) in CF and WT neutrophils were compared. Lastly, specific iNOS inhibitor 1400W was tested to prevent CF intestinal obstruction. Immune restoration in CF mice reversed the intestinal neutrophilic inflammation, improved the intestinal dysmotility, and rescued the mice from mortality. Transfusion of WT neutrophils into CF mice ameliorated the retarded bowel movement. CF neutrophils expressed significantly more iNOS and produced significantly more NO. Pharmaceutical blocking of iNOS significantly improved intestinal transit and survival of CF mice. CF immune defect plays a critical role in CF intestinal disease development. Activation of iNOS in inflammatory cells produces excessive NO, slows the bowel movement, and facilitates intestinal paralysis and obstruction in CF. Thus, normalization of the CF immune system may offer a novel therapy to treat CF intestinal disease.

Effects of dietary succinic acid supplementation on growth performance, digestive ability, intestinal development and immunity of large yellow croaker (Larimichthys crocea) larvae

The application of artificial micro-diet is an effective way to improve and standardize the quality of aquatic animal larvae. However, the widespread adoption of micro-diet faces a bottleneck due to the limited utilization capacity of the larvae. A 30-day feeding experiment was carried out to investigate the effect of dietary succinic acid (SA) on the growth performance, digestive ability, intestinal development, and immunity of large yellow croaker larvae (initial body weight 11.33 ± 0.57 mg). Four isonitrogenous and isolipidic diets were formulated, incorporating 0.00%, 0.01%, 0.02% and 0.03% SA separately. The results showed that a diet with 0.02% SA significantly increased both the final body weight and the specific growth rate of the larvae. Regarding digestive ability, 0.01% SA supplementation significantly enhanced trypsin activity in both intestinal and pancreatic segments. In addition, 0.01% SA supplementation notably improved amylase activity in the intestinal segment, while diets with 0.01%-0.02% SA significantly improved lipase activity in the pancreatic segment. In terms of intestinal development, 0.01% SA supplementation remarkably boosted the activities of alkaline-phosphatase and leucine-aminopeptidase on brush border membrane in intestine. Furthermore, 0.03% SA supplementation significantly increased the expression of occludin. In terms of immunity, larvae fed diets with 0.01%-0.02% SA exhibited significantly higher lysozyme activity compared to the control group. Supplementation with 0.01% SA also significantly increased both iNOS activity and NO content. In summary, the findings of this study suggested that adding 0.02% SA can improve the growth performance of large yellow croaker larvae by improving digestive enzymes activities, promoting intestinal development, and enhancing nonspecific immunity.

Melatonin Mediates Cardiac Tissue Damage under Septic Conditions Induced by Lipopolysaccharide.

Lipopolysaccharide (LPS) is known to induce oxidative stress and inflammation, leading to significant damage in cardiac tissues. This study investigates the protective effects of melatonin (MLT) against LPS-induced oxidative damage, inflammation, and apoptosis in rat heart tissue. Rats were divided into four groups (n = 6 per group): control, melatonin-treated, LPS-treated, and LPS + melatonin-treated. Oxidative stress markers, including thiobarbituric acid-reactive substances (TBARSs) and advanced oxidation protein products (AOPPs), were measured. Additionally, inflammatory markers, such as interleukin-6 (IL-6) levels, inducible nitric oxide synthase (iNOS) and nitric oxide (NO) content, and apoptotic markers, caspase-3, caspase-9, and acidic DNase activity, were evaluated. LPS treatment significantly increased TBARS, AOPP, and IL-6 levels, as well as the activity of caspase-3, acidic DNase and iNOS and NO content compared to the control group. Co-treatment with melatonin significantly reduced the levels of TBARS and AOPP levels, and caspase-3 and acidic DNase activities nearly matched those of the control group, while caspse-9 was still slightly increased. Interestingly, IL-6, iNOS and NO levels were significantly decreased but did not fully match the values in the control group. Melatonin mitigates LPS-induced oxidative stress, inflammation, and apoptosis in rat heart tissue by affecting all studied parameters, demonstrating its potential as a therapeutic agent for conditions characterized by oxidative stress and inflammation. Further research is warranted to explore the clinical applications of melatonin in cardiovascular diseases.

Neuroprotective effect of Bouvardia ternifolia (Cav.) Schltdl via inhibition of TLR4/NF-κB, caspase-3/Bax/Bcl-2 pathways in ischemia/reperfusion injury in rats.

Bouvardia ternifolia is a plant known for its traditional medicinal uses, particularly in treating inflammation and oxidative stress. Recent studies have explored its potential in neuroprotection, especially in the context of cerebral ischemia/reperfusion injury, a condition where blood supply returns to the brain after a period of ischemia, leading to oxidative stress and inflammation. This damage is a major contributor to neuronal death and neurodegenerative diseases. A BCCAO/reperfusion model was induced, followed by treatment with B. ternifolia extract. Various molecular biology methods were employed, including Western blot analysis, gene expression assessment via RT-qPCR, and the measurement of oxidative stress mediators. In the BCCAO/reperfusion model, the compounds in the dichloromethane extract work by targeting various signaling pathways. They prevent the activation of iNOS and nNOS, reducing harmful reactive oxygen and nitrogen species, and boosting antioxidant enzymes like catalase and superoxide dismutase. This lowers oxidative stress and decreases the expression of proteins and genes linked to cell death, such as Bax, Bcl-2, and caspase-3. The extract also blocks the TLR4 receptor, preventing NF-κB from triggering inflammation. Additionally, it reduces the activation of microglia and astrocytes, as shown by lower levels of glial activation genes like GFAP and AiF1. The dichloromethane extract of B. ternifolia demonstrated significant neuroprotective effects in the BCCAO/reperfusion model by modulating multiple signaling pathways. It effectively reduced oxidative stress, inhibited inflammation, and attenuated apoptosis, primarily through the downregulation of key proteins and genes associated with these processes. These findings suggest that the extract holds therapeutic potential for mitigating ischemia/reperfusion-induced neuronal damage.

Effects of L-NAME and air exposure on mitochondrial energetic markers, thyroid hormone receptor/regulator system and stress/ease-responsive receptor expression in the brain/gut axis of zebrafish

As a signal molecule, nitric oxide (NO) has several physiological actions in fish. However, the action of NO on the brain/gut axis, a classic inter-organal axis that bridges the gastrointestinal tract and the CNS, still requires more understanding. The short-term in vivo action of a NO inhibitor, N-omega-nitro-L-arginine methyl ester (L-NAME), on mitochondrial energetic markers and the receptor expression of thyroid hormone (TH) and neuroendocrine hormones involved in stress/ease response was tested in the brain/gut axis of zebrafish exposed to either in non-stressed or air-exposed condition. L-NAME treatment decreased the NO content in brain and gut segments in non-stressed fish but rose upon L-NAME treatment in air-exposed fish that corresponded with the activation of inos, nnos, hif1a and hif1an transcript expressions. The brain/gut segments that showed spatial and differential sensitivity to L-NAME, modified the transcript expression patterns of stress (adra2da, adrb1, nr3c2)- and ease-responsive (htr2b, slc6a4a, mtnr1aa) hormone receptors. The expression pattern of the TH receptor/regulator system (thra, thrb, dio1, dio2, dio3) becomes more active in gut segments than brain segments upon L-NAME challenge in stressed zebrafish. The data provide evidence for a novel role of NO as an integrator of brain/gut axis segments in zebrafish, where the endogenously produced NO in mid-brain/posterior-gut axis aligns together upon air-exposure stress, providing a lead role to the posterior gut that activates and directs the neuroendocrine receptor expressions of stress/ease responsive genes. The data further invites studies exploring the therapeutic potential of L-NAME in this biomedical model to control the brain/gut axis segments.

Sulfate-Reducing Bacteria Induce Pro-Inflammatory TNF-α and iNOS via PI3K/Akt Pathway in a TLR 2-Dependent Manner.

Desulfovibrio, resident gut sulfate-reducing bacteria (SRB), are found to overgrow in diseases such as inflammatory bowel disease and Parkinson's disease. They activate a pro-inflammatory response, suggesting that Desulfovibrio may play a causal role in inflammation. Class I phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway regulates key events in the inflammatory response to infection. Dysfunctional PI3K/Akt signaling is linked to numerous diseases. Bacterial-induced PI3K/Akt pathway may be activated downstream of toll-like receptor (TLR) signaling. Here, we tested the hypothesis that Desulfovibrio vulgaris (DSV) may induce tumor necrosis factor alpha (TNF-α) and inducible nitric oxide synthase (iNOS) expression via PI3K/Akt in a TLR 2-dependent manner. RAW 264.7 macrophages were infected with DSV, and protein expression of p-Akt, p-p70S6K, p-NF-κB, p-IkB, TNF-α, and iNOS was measured. We found that DSV induced these proteins in a time-dependent manner. Heat-killed and live DSV, but not bacterial culture supernatant or a probiotic Lactobacillus plantarum, significantly caused PI3K/AKT/TNF/iNOS activation. LY294002, a PI3K/Akt signaling inhibitor, and TL2-C29, a TLR 2 antagonist, inhibited DSV-induced PI3K/AKT pathway. Thus, DSV induces pro-inflammatory TNF-α and iNOS via PI3K/Akt pathway in a TLR 2-dependent manner. Taken together, our study identifies a novel mechanism by which SRB such as Desulfovibrio may trigger inflammation in diseases associated with SRB overgrowth.

Зміни концентрації sST2, активності мієлопероксидази та iндуцибельної NO-синтази в сироватці крові та лімфоцитах як показник важкості перебігу захворювання у пацієнтів з артеріальною гіпертензією та на тлі коронавірусної хвороби

The prevalence of COVID-19 and its polymorphic clinical manifestations are attributed to a systemic inflammatory response, which also plays a key role in the development of arterial hypertension (AH). The prognosis and effectiveness of treatment in patients with AH and COVID-19 should be assessed based on the levels of inflammatory biomarkers sST2, myeloperoxidase, and iNOS. The aim of the work was to analyze the prognostic value sST2, myeloperoxidase and iNOS levels in the blood serum of patients with arterial hypertension (AH) and COVID-19. Methods. 2 groups of patients were examined: group 1- 36 patients with AH and hypertensive crisis. Group 2 – 35 patients with AH and polysegmental pneumonia on the background of COVID-19. The control group – 16 practically healthy individuals. All patients underwent anthropometry, determination of biochemical blood tests, echocardiography, level of sST2, and activity of iNOS and MPO using ELISA in blood serum and lymphocytes. Results. A 2.4-fold increase in sST2 content in blood serum was noted in AH and 2.9-fold in the background of COVID-19. The level of myeloperoxidase in blood serum increased 2.5 times in hypertension and 3.4 times in coronavirus disease. In lymphocytes, iNOS activity increased 3.25 times in hypertension and 4.3 times in COVID-19. sST2 level has a significant correlation with the size of the left atrium, left ventricle, and ejection fraction in patients with AH. A positive correlation with age was noted in the group of patients with AH and COVID-19. Conclusions. Levels of sST2, iNOS and myeloperoxidase are important markers of the severity of cardiovascular pathology, as well as in COVID-19 patients. In patients with arterial hypertension and in combination with COVID-19, a significant increase in the concentration of sST2, myeloperoxidase and iNOS was observed compared to practically healthy individuals. A significant elevation in myeloperoxidase levels has been noted in patients with AH without coronavirus disease, indicating the utility of its use as a highly sensitive marker for low-intensity inflammation, particularly in arterial hypertension. This makes this marker more promising for use than C-reactive protein.

Exposure to polystyrene nanoplastics causes immune damage, oxidative stress and intestinal flora disruption in salamander (Andrias davidianus) larvae

The toxic effects of nanoparticles have been increasingly investigated, but there has been limited research on amphibians, especially those of conservation value. This study examined the effects of different concentrations (0, 0.04, 0.2, 1, 5 mg/L) of polystyrene nanoplastics (PS-NPs, 80 nm) on the short-term exposure (7 d) of Andrias davidianus. Results demonstrated the concentration-dependent enrichment of PS-NPs in the intestine. Histological lesions displayed increased hepatic macrophages with cellular rupture, broken intestinal villi, decreased cuprocytes and crypt depression. Antioxidant- and inflammation-related enzyme activities were analysed, and it was found that hepatic and intestinal MDA content and CAT activity were highest in the N-1 group and SOD activity was highest in the N-0.2 group (p < 0.05). AKP activity continued to decline, and iNOS activity was highest in the N-0.2 group (p < 0.05). il-10, tgf-β, bcl-w and txnl1 were significantly downregulated in the N-0.2 group, while il-6 and il-8 were markedly upregulated in the N-0.2 group (p < 0.05). Exposing to PS-NPs decreased probiotic bacteria (Cetobacterium, Akkermansia) and increased pathogenic bacteria (Lachnoclostridium). Our results suggest that NPs exposure can have deleterious effects on salamanders, which predicts that NPs contamination may lead to continued amphibian declines. Therefore, we strongly recommend that attention be paid to amphibians, especially endangered species, in the field of NPs.

Pioglitazone ameliorates sepsis-associated encephalopathy through SIRT1 signaling pathway

Sepsis is a severe immune response to an infection. It is associated with multiple organ dysfunction syndrome (MODs) along with systemic and neuronal inflammatory response. This study focused on the acute neurologic dysfunction associated with sepsis by exploring the role of PPARγ/SIRT1 pathway against sepsis. We studied the role of this axis in ameliorating sepsis-associated encephalopathy (SAE) and its linked neurobehavioral disorders by using pioglitazone (PIO). This PPARγ agonist showed neuroprotective actions in neuroinflammatory disorders. Sepsis was induced in mice by LPS (10 mg/kg). Survival rate and MODs were assessed. Furthermore, behavioral deficits, cerebral oxidative, inflammatory, and apoptotic markers, and the cerebral expression level of SIRT1 were determined. In this study, we observed that PIO attenuated sepsis-induced cerebral injury. PIO significantly enhanced survival rate, attenuated MODs, and systemic inflammatory response in septic mice. PIO also promoted cerebral SIRT1 expression and reduced cerebral activation of microglia, oxidative stress, HMGB, iNOS, NLRP3 and caspase-3 along with an obvious improvement in behavioral deficits and cerebral pathological damage induced by LPS. Most of the neuroprotective effects of PIO were abolished by EX-527, a SIRT1 inhibitor. These results highlight that the neuroprotective effect of PIO in SAE is mainly SIRT1-dependent.

Fucoidan from Laminaria japonica protects renal tubular epithelial cells from uric acid induced NLRP3-mediated pyroptosis through inhibition of NF-κB pathway

Ethnopharmacological relevanceHyperuricemia is a common metabolic disease with prominent morbidity, it can lead to many adverse effects and complications, such as chronic nephrosis. Fucoidan has been used as natural drug for acute and chronic kidney disease for over 20 years in China, but the precise mechanisms underlying the renal protective function are still indefinable. PurposeThis study is conducted to explore alleviation of fucoidan (FPS) from Laminaria japonica on urate-induced NOD-like receptor family, pyrin domain-containing 3 (NLRP3)-mediated pyroptosis in renal tubular epithelial cells HK-2, as well as the mechanism of nuclear factor κB (NF-κB) signaling pathway involved. Materials and methodsHK-2 cells were treated with FPS, uric acid (UA), and inhibitor of NF-κB signaling pathway. Nitric oxide (NO) content and inducible nitric oxide synthase (iNOS) activity were determined with detection kits. Activation of intercellular NLRP3 inflammasome and NF-κB signaling pathway, gasdermin D (GSDMD) expression level were evaluated with Western blot and quantitative reverse transcription-PCR (qRT-PCR), and immunofluorescent analysis. ResultsData showed that UA induced cellular inflammatory response demonstrated by elevated NO content, iNOS activity and expression level of NLRP3 inflammasome-mediated pyroptosis associated molecules including NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), Caspase-1, interleukin 18 (IL-18) and GSDMD, moreover the NF-κB signaling pathway was activated by UA. However, FPS exposure inhibited efficiently the UA induced adverse effect. ConclusionIt can be concluded that FPS inhibited UA-induced NLRP3-mediated pyroptosis in HK-2 cells through repressing NF-κB signaling pathway.

Cyclic Nitroxide 4-Methoxy-Tempo May Decrease Serum Amyloid A-Mediated Renal Fibrosis and Reorganise Collagen Networks in Aortic Plaque.

Acute-phase serum amyloid A (SAA) can disrupt vascular homeostasis and is elevated in subjects with diabetes, cardiovascular disease, and rheumatoid arthritis. Cyclic nitroxides (e.g., Tempo) are a class of piperidines that inhibit oxidative stress and inflammation. This study examined whether 4-methoxy-Tempo (4-MetT) inhibits SAA-mediated vascular and renal dysfunction. Acetylcholine-mediated vascular relaxation and aortic guanosine-3',5'-cyclic monophosphate (cGMP) levels both diminished in the presence of SAA. 4-MetT dose-dependently restored vascular function with corresponding increases in cGMP. Next, male ApoE-deficient mice were administered a vehicle (control, 100 µL PBS) or recombinant SAA (100 µL, 120 µg/mL) ± 4-MetT (at 15 mg/kg body weight via i.p. injection) with the nitroxide administered before (prophylaxis) or after (therapeutic) SAA. Kidney and hearts were harvested at 4 or 16 weeks post SAA administration. Renal inflammation increased 4 weeks after SAA treatment, as judged by the upregulation of IFN-γ and concomitant increases in iNOS, p38MAPK, and matrix metalloproteinase (MMP) activities and increased renal fibrosis (Picrosirius red staining) in the same kidneys. Aortic root lesions assessed at 16 weeks revealed that SAA enhanced lesion size (vs. control; p < 0.05), with plaque presenting with a diffuse fibrous cap (compared to the corresponding aortic root from control and 4-MetT groups). The extent of renal dysfunction and aortic lesion size was largely unchanged in 4-MetT-supplemented mice, although renal fibrosis diminished at 16 weeks, and aortic lesions presented with redistributed collagen networks. These outcomes indicate that SAA stimulates renal dysfunction through promoting the IFN-γ-iNOS-p38MAPK axis, manifesting as renal damage and enhanced atherosclerotic lesions, while supplementation with 4-MetT only affected some of these pathological changes.

Effect of lipopolysaccharide on the development of oxidativenitrosative stress in salivary glands and soft periodontal tissues of rats under conditions of water avoidance stress

Introduction and aim. Violation in the salivary glands will inevitably cause changes in periodontium, and periodontitis can disrupt the functioning of the salivary glands. The purpose of the work is to evaluate changes in NO-synthase and arginase activities, pro- and antioxidant balance in rat salivary glands and soft periodontal tissues during administration of bacterial lipopolysaccharide (LPS) and water avoidance stress (WAS) modeling. Material and methods. The experiment was performed on 24 rats. The animals were divided into 4 groups: control, WAS, animals injected intraperitoneally with 0.4 μg/kg of bacterial LPS of Salmonella typhi, WAS+LPS. Results. Water avoidance stress led to decrease of inducible NO-synthase (iNOS) activity in salivary glands by 1.63 times, but decreased arginase activity by 1.15 times, superoxide production increased by 1.53 times, catalase activity decreased by 1.2 times, and malonic dialdehyde (MDA) increased by 1.19 times compared to the control. Lipopolysaccharide led to increase of constitutive NO-synthases (cNOS) activity in salivary glands by 1.48 times, but decreased arginase activity by 6.15 times, catalase activity increased by 2.6 times and superoxide dismutase activity decreased by 2.74 times, and MDA increased by 6.84 times compared to the control. Water avoidance stress + LPS in salivary glands led to decrease of cNOS and arginase activity by 1.09 and 1.19 times, respectively, superoxide production increased by 1.88 times, catalase and superoxide dismutase activity decreased by 1.06 times and 1.34 times, respectively, and MDA increased by 2.44 times compared to the control. Water avoidance stress led to increase of iNOS activity in periodontium by 1.44 times and arginase activity decreased by 1.37 times, superoxide production increased 1.32 times, catalase activity and superoxide dismutase activities decreased by 1.27 times and by 1.53 times, respectively, and MDA increased by 1.31 times compared to the control. Lipopolysaccharide led to increase of iNOS activity in the periodontium by 3.88 times, arginase activity decreased by 2.69 times, superoxide production increased 1.64 times, catalase activity increased by 4.32 times, and MDA increased by 4.51 times compared to the control. Water avoidance stress + LPS in periodontium led to increase of iNOS and cNOS activities by 1.95 times and 1.53, respectively, arginase activity decreased by 1.39 times, superoxide production increased 1.66 times, catalase activity increased by 1.11 times, and MDA increased by 1.53 times compared to the control. Conclusion. The combination of LPS and WAS leads to changes in NO production and oxidative stress in salivary glands and the periodontium.

Phytochemical analysis of Cichorium bottae root and anti-inflammatory potential assessment of isolates

The Cichorium plants are particularly notable due to their remarkable therapeutic and medicinal properties, besides being used as food and conventional medication. Although Cichorium plants have been studied for their phytoconstituents and biological activities, there is limited knowledge about the constituents of the roots of C. bottae. A phytochemical study of the 90% MeOH extract of C. bottae roots resulted in the isolation of twelve compounds belonging to guaianolide sesquiterpene lactones, sesquiterpene lactone glucosides, and phenolic derivatives, of which two compounds designated as 9α-hydroxycrepediaside B (1) and cichobotinal (2) were previously undescribed. The isolated compounds were assessed for their anti-inflammatory potential through the inhibition of inducible nitric oxide synthase (iNOS) and resultant decrease in nitric oxide generation in LPS-induced macrophages. Among the isolates, compounds 2 and 11 (8-deoxylactucin) inhibited iNOS activity with IC50 values of 21.0 ± 4 and 6.8 ± 0.1 μM, respectively. The methanolic extract of C. bottae inhibited iNOS with an IC50 of 10.5 ± 0.5 μg/mL.

Early Socialization Triggered ROS-Mediated Activation of Canonical NF-κB Pathway Leading to Inflammation of Spleen in Suckling Piglets

Early socialization during lactation is advocated as a feeding strategy to reduce the weaning stress of piglets. However, early socialization has often been accompanied by more frequent aggression between individuals, and its effect on the immune system of piglets has yet to be evaluated. In this study, 89 piglets were raised separately under conventional feeding and early socialization environments. Based on differences in the aggressive behavior of the piglets in different environments during lactation, we further investigated the effects of early socialization on oxidative stress in the spleen of the piglets and the inflammatory responses involved in the canonical nuclear factor kappa-B (NF-κB) signaling pathway. The results revealed that early socialization led to a higher aggression level between individuals (p &lt; 0.01), increased malondialdehyde (MDA) and H2O2 levels and inducible nitric oxide synthase (iNOS) activity, and inhibited glutathione (GSH) levels and the activities of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPX) in the piglet spleens (p &lt; 0.05). The mRNA expression levels of the protein kinase A (PKA), inhibitor of kappa B kinase-α (IKK-α), inhibitor of kappa B kinase-β (IKK-β), inhibitor of NF-κB-α (IκB-α), NF-κB(p65), interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (COX2), iNOS, and heat shock protein (HSP) genes were significantly up-regulated, as well as the protein levels of P-p65, IKK-β, P-IkB-α, pro-IL-1β, and TNF-α. In summary, early socialization caused oxidative stress and inflammatory responses in the spleen of the piglets by inducing ROS production and the activation of the canonical NF-κB pathway. Our study revealed that early socialization significantly increased the ROS level in the piglet spleens and activated the canonical NF-κB signaling pathway, which induced a high expression of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, and COX2) and HSP genes regulated by NF-κB signaling, leading to oxidative stress and the inflammatory response.

The influence of monosodium glutamate on the development of oxidative-nitrosative stress in the large cerebral hemispheres of rats under the combination of a change in the “light-dark” cycle and a systemic inflammatory response

Today, the issue of the consequences of disrupting the normal “light-dark” cycle is becoming increasingly important, and the link between the development of a systemic inflammatory response (SIR) and circadian rhythm disorders has been confirmed. The effect of monosodium glutamate on the human body is being actively studied. The aim of the work was to find out the influence of monosodium glutamate on the development of oxidative-nitrosative stress in the homogenate of the large cerebral hemispheres of rats with a combination of acute desynchronosis (AD), SIR and administration of monosodium glutamate. Material and methods. The research was carried out on 72 white Wistar rats weighing 150–200 of different sexes, divided into 5 groups: control (n = 15), AD (n = 13), SIR (n = 15), a combination of SIR and AD (n = 14), a combination of SIR, AD, and glutamate (n = 15). To simulate AD, a normal “light-dark” cycle (12 hours of light, 12 hours of darkness) was formed for 3 weeks, and the next 3 days the “light-dark” phases were shifted back by 6 hours. SIR was reproduced by intraperitoneal injection of Salmonella typhi lipopolysaccharide in the first week at a dose of 0.4 μg/kg 3 times per week, the following seven weeks – once a week. Sodium glutamate was administered intragastrically for 20 days at a dose 30 mg/kg, dissolved in 0.5 ml of distilled water. In a 10 % homogenate of the large cerebral hemispheres, we determined the rate of superoxide anion radical (SAR) production, the content of products that react with thiobarbituric acid (TBA-reactants), their increase, activity of catalase, superoxide dismutase (SOD), ornithine decarboxylase (ODC), concentration of nitrites, peroxynitrites, total activity of NO-synthase (NOS), its constitutive (cNOS) and inducible (iNOS) isoforms. Results. In the group of SIR, AD and glutamate combination, compared to the control, the following was noted: an increase in the rate of basic production of SAR by 175.8 %, by the microsomal pathway by 20.0 %, by the mitochondrial pathway by 51.2 %, the concentration of TBA-reactants by 83.4 %, their increase – by 61.7 %, decrease in the activity of SOD by 57.1 %, catalase – by 38.1 %; an increase in peroxynitrite content by 116.7 %, ODC activity by 161.5 %, total NOS activity by 25.6 % and iNOS by 27.4 %, a decrease in cNOS activity by 15.0 % and nitrite concentration by 35.0 % in brain tissues. Conclusions. Sodium glutamate administration in combination with AD and SIR leads to increased oxidative-nitrosative stress, inhibition of antioxidant protection; contributes to the activation of the general activity of NO-synthase and iNOS, suppresses the activity of cNOS.

Hesperetin alleviates pulmonary injury in a blunt chest trauma-induced pulmonary contusion model in rats

Pulmonary contusion (PC), a condition that occurs frequently in severe thoracic injuries, is a significant contributor to mortality in those under the age of 40. Hesperetin, a natural flavonoid derivative of hesperidin, is a substance found in various citrus fruits such as oranges and grapefruits and possesses a variety of biological activities, including antiapoptotic, antioxidant, and anticancer effects. In the current study, we investigated the effect of hesperetin on pulmonary tissue structure, expression of some pro-inflammatory cytokines, and mediators in a PC model induced by blunt chest trauma. In this study, 18 adult male Wistar albino rats (8 &amp;ndash; 10 weeks, 250 &amp;ndash; 300 g) were used. The rats were divided into three groups: control, PC, and PC + hesperetin. Hesperetin administration (100 mg/kg/day oral) was completed for 7 days following induction of the model. The wet/dry weight ratio of pulmonary tissue was determined. Tumor necrosis factor alpha (TNF&amp;alpha;) and malondialdehyde (MDA) in lung tissues, serum interleukin (IL)-6, and IL-1&amp;beta; levels were determined using the enzyme-linked immunosorbent assays. Pulmonary tissue specimens were examined histologically using hematoxylin-eosin and Masson trichrome staining. Inducible nitric oxide synthase (iNOS) activity was determined using immunohistochemical methods. Hesperetin administration reduced TNF&amp;alpha; and iNOS activity, serum IL-1&amp;beta;, IL-6, MDA, and wet/dry weight ratio in pulmonary tissue to improve pulmonary function. Our results showed that administration of hesperetin prevents activation of local inflammatory mediators, thereby obstructing the proinflammatory cytokine cascade and tissue injury.

Metabolomic Profiling and In Vivo Antiepileptic Effect of Zygophyllum album Aerial Parts and Roots Crude Extracts against Pentylenetetrazole-Induced Kindling in Mice.

The chemical profiles of both Zygophyllum album (Z. album) aerial parts and roots extracts were evaluated with LC-ESI-TOF-MS/MS analysis. Twenty-four compounds were detected. Among them, some are detected in both the aerial parts and the roots extracts, and others were detected in the aerial parts only. The detected compounds were mainly flavonoids, phenolic compounds, triterpenes and other miscellaneous compounds. Such compounds contribute to the diverse pharmacological activities elicited by the Z. album species. This study aimed to elucidate the antiepileptic effect of Z. album aerial parts and roots crude extracts against pentylenetetrazole (PTZ)-induced kindling in mice. Male albino mice were divided into four groups, eight animals each. All groups, except the control group, were kindled with PTZ (35 mg/kg i.p.), once every alternate day for a total of 15 injections. One group was left untreated (PTZ group). The remaining two groups were treated prior to PTZ injection with either Z. album aerial parts or roots crude extract (400 mg/kg, orally). Pretreatment with either extract significantly reduced the seizure scores, partially reversed the histological changes in the cerebral cortex and exerted antioxidant/anti-inflammatory efficacy evinced by elevated hippocampal total antioxidant capacity and SOD and catalase activities, parallel to the decrement in MDA content, iNOS activity and the TXNIB/NLRP3 axis with a subsequent decrease in caspase 1 activation and a release of IL-1β and IL-18. Moreover, both Z. album extracts suppressed neuronal apoptosis via upregulating Bcl-2 expression and downregulating that of Bax, indicating their neuroprotective and antiepileptic potential. Importantly, the aerial parts extract elicited much more antiepileptic potential than the roots extract did.

Prdx1 regulates macrophage polarization by maintaining mitochondrial homeostasis

In order to investigate the role of Prdx1 in macrophage polarization, mouse leukemia cells of monocyte macrophage (RAW264.7) were treated with lipopolysaccharides (LPS)+ interferon gamma (IFNγ) or IL-4 to induce type 1 macrophage (M1) and type 1 macrophage (M2) macrophages, respectively. The Prdx1 gene knockout cells (Prdx1-/-) were used for the study. Flow cytometry was conducted to detect M1/M2 macrophage markers, and ELISA kits were used to measure M1/M2 cytokine levels. Inducible nitric-oxide synthase (iNOS) activity, arginase-1 (Arg-1) activity, and oxidative damage were also assessed. The Seahorse XFe24 Extracellular Flux Analyzer was employed to measure extracellular acidification rate and oxygen consumption rate. The mitochondrial membrane potential was analyzed using the mitochondrial membrane potential dye (JC-1) fluorescent probe, and mitochondrial superoxide was detected through fluorescence staining. Additionally, the impact of adding a mitochondrial reactive oxygen species (ROS) scavenger on RAW264.7 macrophage polarization was examined. The results demonstrated an increase in ROS, hydrogen peroxide, and 8-hydroxy-2 deoxyguanosine (8-OHDG). Cytotoxicity and mitochondrial toxic effects, including mitochondrial superoxide accumulation, decreased adenosine-triphosphate (ATP) production, reduced mitochondrial membrane potential, and decreased mitochondrial DNA copy number, were observed. Furthermore, down-regulation of translocase of inner mitochondrial membrane 23 (TIM23) mitochondrial protein and mitochondrial stress protein heat shock protein 60 (HSP60) was noted. The extra cellular acidification rate (ECAR) in M1 macrophage polarization in RAW264.7 cells was increased, while oxygen consumption rate (OCR) in M2 macrophages was reduced. These findings indicate that Prdx1 knockout in RAW264.7 cells can inhibit M2 macrophage polarization but promote M1 macrophage polarization by impairing mitochondrial function and reducing oxidative phosphorylation.

The effects of ecdysterone and enalapril on nitric oxide synthesis and the markers of oxidative stress in streptozotocin-induced diabetes in rats: a comparative study.

Cardiovascular functions in diabetes greatly depend on constitutive NOS (cNOS) activity. A comparative study of the effects of a steroid hormone ecdysterone and enalapril, an ACE inhibitor widely used to treat cardiac disorders on cNOS, inducible NOS (iNOS), xanthine oxidoreductase (XOR) activity, RNS, ROS, and lipid peroxidation in heart tissue in experimental diabetes was conducted. The rat model of diabetes was established by streptozotocin injection. NOS activity, NO2-, NO3-, uric acid, nitrosothiols, hydroperoxide, superoxide, and diene conjugate formation were studied spectrophotomerically. In diabetes, cNOS downregulation correlated with a dramatic fall of NO2- production and ~4.5-fold elevation of nitrosothiols, which agreed with a steep rise of iNOS activity, while NO3- remained close to control. Dramatic activation of XOR was observed, which correlated with the elevation of both superoxide production and nitrate reductase activity and resulted in strong lipid peroxidation. Ecdysterone and enalapril differently affected RNS metabolism. Ecdysterone moderately restored cNOS but strongly suppressed iNOS, which resulted in the reduction of NO3-, but full restoration of NO2- production. Enalapril better restored cNOS but less effectively suppressed iNOS, which promoted NO3- formation. Both drugs similarly inhibited XOR, which equally alleviated oxidative stress and lipid peroxidation. The synergistic action of iNOS and XOR was a plausible explanation for strong lipid peroxidation, abolished by the inhibition of iNOS and XOR by ecdysterone or enalapril. Complementary effects of ecdysterone and enalapril on cNOS, iNOS, and RNS are a promising basis for their combined use in the treatment of cardiovascular disorders caused by cNOS dysfunction in diabetes.