Nitric Oxide Activity Research Articles

Effect of increasing ambient temperature on physiological changes, oxidative stress, nitric oxide, total antioxidant power, and mitochondrial activity of Nile tilapia (Oreochromis niloticus Linn.)

The objective of this study was to investigate the effect of high ambient temperature on physiological changes of live Nile tilapia (Oreochromis niloticus Linn.) and biochemical parameters in Nile tilapia blood cells. Live Nile tilapia and Nile tilapia blood cells were maintained at temperature ranges from 24 to 34 °C and 24–38 °C, respectively. Then, physiological parameters (body temperature, body color score, pulse rate, ventilation rate, oxygen saturation) of the live Nile tilapia and biochemical parameters (total antioxidant power, hydrogen peroxide, malondialdehyde, nitric oxide, and mitochondrial activity) of the Nile tilapia blood cells were measured for every 2 °C increase. It was found that when the ambient temperature increases, body temperature of live Nile tilapia continually increased. Total antioxidant power and hydrogen peroxide, pulse rate, body color score, malondialdehyde, ventilation rate and mitochondrial activity commenced to increase at 26, 28, 30, 32, 34 °C (P < 0.05), respectively. Blood oxygen saturation decreased at 32 °C (P < 0.05). The response of nitric oxide of Nile tilapia blood cells responded to increased temperature in two phases. In the first phase, nitric oxide decreased at 28–32 °C (P < 0.05), then increased and decreased again at a temperature of 38 °C (P < 0.05). These phenomena indicated that when the live Nile tilapias and the Nile tilapia blood cells were exposed to high ambient temperature, they were under heat stress and oxidative stress. In addition, physiological responses in the live Nile tilapia and biochemical changes in their blood cells when exposed to high ambient temperature occurred in a consistent manner when the temperature continually increased.

Mitochondrial CypD Acetylation Promotes Endothelial Dysfunction and Hypertension.

Nearly half of adults have hypertension, a major risk factor for cardiovascular disease. Mitochondrial hyperacetylation is linked to hypertension, but the role of acetylation of specific proteins is not clear. We hypothesized that acetylation of mitochondrial CypD (cyclophilin D) at K166 contributes to endothelial dysfunction and hypertension. To test this hypothesis, we studied CypD acetylation in patients with essential hypertension, defined a pathogenic role of CypD acetylation in deacetylation mimetic CypD-K166R mutant mice and endothelial-specific GCN5L1 (general control of amino acid synthesis 5 like 1)-deficient mice using an Ang II (angiotensin II) model of hypertension. Arterioles from hypertensive patients had 280% higher CypD acetylation coupled with reduced Sirt3 (sirtuin 3) and increased GCN5L1 levels. GCN5L1 regulates mitochondrial protein acetylation and promotes CypD acetylation, which is counteracted by mitochondrial deacetylase Sirt3. In human aortic endothelial cells, GCN5L1 depletion prevents superoxide overproduction. Deacetylation mimetic CypD-K166R mice were protected from vascular oxidative stress, endothelial dysfunction, and Ang II-induced hypertension. Ang II-induced hypertension increased mitochondrial GCN5L1 and reduced Sirt3 levels resulting in a 250% increase in GCN5L1/Sirt3 ratio promoting CypD acetylation. Treatment with mitochondria-targeted scavenger of cytotoxic isolevuglandins (mito2HOBA) normalized GCN5L1/Sirt3 ratio, reduced CypD acetylation, and attenuated hypertension. The role of mitochondrial acetyltransferase GCN5L1 in the endothelial function was tested in endothelial-specific GCN5L1 knockout mice. Depletion of endothelial GCN5L1 prevented Ang II-induced mitochondrial oxidative stress, reduced the maladaptive switch of vascular metabolism to glycolysis, prevented inactivation of endothelial nitric oxide, preserved endothelial-dependent relaxation, and attenuated hypertension. These data support the pathogenic role of CypD acetylation in endothelial dysfunction and hypertension. We suggest that targeting cytotoxic mitochondrial isolevuglandins and GCN5L1 reduces CypD acetylation, which may be beneficial in cardiovascular disease.

Retraction Note: Gastroprotective effect of desmosdumotin C isolated from Mitrella kentii against ethanol-induced gastric mucosal hemorrhage in rats: possible involvement of glutathione, heat-shock protein-70, sulfhydryl compounds, nitric oxide, and anti-Helicobacter pylori activity.

Retraction Note: Gastroprotective effect of desmosdumotin C isolated from Mitrella kentii against ethanol-induced gastric mucosal hemorrhage in rats: possible involvement of glutathione, heat-shock protein-70, sulfhydryl compounds, nitric oxide, and anti-Helicobacter pylori activity.

Caffeic acid attenuates memory dysfunction and restores the altered activity of cholinergic, monoaminergic and purinergic in brain of cadmium chloride exposure rats.

This study aims to evaluate the neuroprotective effect of caffeic acid (CAF) against cadmium chloride (CdCl2) in rats via its effect on memory index as well as on altered enzymatic activity in the brain of CdCl2-induced neurotoxicity. The experimental rats were divided into seven groups (n=6 rats per group) of healthy rats (group 1), CdCl2 -induced (CD) (3 mg/kg BW) rats (group 2), CD rats + Vitamin C (group 3), CD rats + CAF (10 and 20 mg/kg BW respectively) (group 4 & 5), and healthy rat + CAF (10 and 20 mg/kg BW respectively) (group 6 & 7). Thereafter, CdCl2 and CAF were administered orally to the experimental rats in group 2 to group 5 on daily basis for 14days. Then, the Y-maze test was performed on theexperimental rats to ascertain their memory index. CdCl2 administration significantly altered cognitive function, the activity of cholinesterase, monoamine oxidase, arginase, purinergic enzymes, nitric oxide (NOx), and antioxidant status of Cd rats (untreated) when compared with healthy rats. Thereafter, CD rats treated with vitamin C and CAF (10 and 20 mg/kg BW) respectively exhibited an improved cognitive function, and the observed altered activity of cholinesterase, monoamine oxidase, arginase, purinergic were restored when compared with untreated CD rats. Also, the level of brain NOx and antioxidant status were significantly (p<0.05) enhanced when compared with untreated CD rats. In the same vein, CAF administration offers neuro-protective effect in healthy ratsvis-à-vis improved cognitive function, reduction in the activity of some enzymes linked to the progression of cognitive dysfunction, and improved antioxidant status when compared to healthy rats devoid of CAF. This study demonstrated the neuroprotective effect of CAF against CdCl2 exposure and in healthy rats.

β-Cell-selective regulation of gene expression by nitric oxide.

Nitric oxide is produced at low micromolar levels following the induction of inducible nitric oxide synthase (iNOS) and is responsible for mediating the inhibitory actions of cytokines on glucose-stimulated insulin secretion by islets of Langerhans. It is through the inhibition of mitochondrial oxidative metabolism, specifically aconitase and complex 4 of the electron transport chain, that nitric oxide inhibits insulin secretion. Nitric oxide also attenuates protein synthesis, induces DNA damage, activates DNA repair pathways, and stimulates stress responses (unfolded protein and heat shock) in β-cells. In this report, the time- and concentration-dependent effects of nitric oxide on the expression of six genes known to participate in the response of β-cells to this free radical were examined. The genes included Gadd45α (DNA repair), Puma (apoptosis), Hmox1 (antioxidant defense), Hsp70 (heat shock), Chop (UPR), and Ppargc1α (mitochondrial biogenesis). We show that nitric oxide stimulates β-cell gene expression in a narrow concentration range of ∼0.5-1 µM or levels corresponding to iNOS-derived nitric oxide. At concentrations greater than 1 µM, nitric oxide fails to stimulate gene expression in β-cells, and this is associated with the inhibition of mitochondrial oxidative metabolism. This narrow concentration range of responses is β-cell selective, as the actions of nitric oxide in non-β-cells (α-cells, mouse embryonic fibroblasts, and macrophages) are concentration dependent. Our findings suggest that β-cells respond to a narrow concentration range of nitric oxide that is consistent with the levels produced following iNOS induction, and that these concentration-dependent actions are selective for insulin-containing cells.

Pathophysiological basis of hepatopulmonary syndrome

Circulatory changes with increased blood flow and vasodilatation/vasoconstriction imbalance are an integral consequence of liver cirrhosis and portal hypertension and can affect the pulmonary circulation with the development of vascular disorders, with hepatopulmonary syndrome (HPS) being the most common. HPS is a serious pulmonary complication of progressive liver disease, resulting in a poor clinical prognosis. Vascular tone decrease, monocytic infiltration of pulmonary vessels, formation of intrapulmonary arteriovenous shunts, dysfunction of alveolar type II cells, destruction of the endothelial glycocalyx are important in the pathogenesis of HPS. Abnormalities of pulmonary capillaries lead to hypoxemia caused by a violation of the ventilation/perfusion ratio, diffusion disorders, and the development of arteriovenous anastomoses. Infiltration of the pulmonary vessels by monocytes is one of the key factors of HPS. This migration is facilitated by the intestinal microbiota translocation into the portal bloodstream with increased expression of proinflammatory cytokines (tumor necrosis factor α, interleu­kins 1, 6), leading to the activation of monocytes. Monocytes located in the pulmonary circulation promote the vasodilation through the activation of inducible nitric oxide (NO) synthase and thus NO production. This is also associated with endothelial dysfunction due to a decreased hepatic secretion of bone morphogenetic protein 9 and increased endothelin 1, endothelial overexpression of endothelin B receptors, and increased endothelial NO production. Proangiogenic factors such as vascular endothelial growth factor, platelet-derived growth factor, and placental growth factor play an important role in the proliferation of pulmonary capillaries. Circulation of tumor necrosis factor α, bile acids and monocyte infiltration in the pulmonary circulation lead to increased apoptosis of alveolar type II cells and decreased surfactant synthesis. Chronic inflammation in HPS disrupts the continuity of the endothelial glycocalyx layer. This article provides an overview of the current knowledge on the pathogenesis of HPS, summarizes many features of the disease based on the literature research in MEDLINE database on the PubMed platform.

A combination of generated hydrogen sulfide and nitric oxide activity has a potentiated protectant effect against cisplatin induced nephrotoxicity

AimHydrogen sulfide and nitricoxide possess cytoprotective activity and in vivo, they are generated from exogenous sodium hydrosulfide and L-arginine respectively. Cisplatin is a major chemotherapeutic agent used to treat cancer and has a high incidence of nephrotoxicity as a side effect. The study aim was to explore the effects of NaHS and L-arginine or their combination on cisplatin induced nephrotoxicity in rats. MethodsWistar Kyoto rats were given a single intraperitoneal dose of cisplatin (5 mg/kg) followed either by NaHS (56 μmol/kg, i. p.), L-arginine (1.25 g/L in drinking water) or their combination daily for 28-days. Post-mortem plasma, urine and kidney samples were collected for biochemical assays and histopathological analysis. ResultsCisplatin decreased body weights and increased urinary output, while plasma creatinine and urea levels were elevated, but sodium and potassium concentrations were diminished. The renal function parameters, blood urea nitrogen and creatinine clearance, were raised and decreased respectively. Regarding markers of reactive oxygen species, plasma total superoxide dismutase was reduced, whereas malondiadehyde was augmented.Cisplatin also diminished plasma and urinary H2S as well as plasma NO, while NaHS and L-arginine counteracted this activity on both redox-active molecules. Cisplatin cotreatment with NaHS, and/or L-arginine exhibited a reversal of all other measured parameters. ConclusionIn current study, NaHS and L-arginine as monotherapy protected the rats from cisplatin-induced nephrotoxicity but the combination of both worked more effectively suggesting the augmented anti-inflammatory and antioxidative potential of test treatments when administered together.

Therapeutic Potential of Rosa canina L. Seed Oil and Metformin: Modulation of Bax/Bcl-2/p53/Caspase-3 Pathways and Improvement of Biochemical and Oxidative Stress Parameters in Estradiol-induced Polycystic Ovary Syndrome Rat Model

Background: This study investigated the therapeutic effects of Rosa canina L. seed oil (RCSO) on PCOS using an animal model, focusing on evaluating biochemical factors and the Bax/Bcl-2/p53 and Caspase-3 (Cas-3) pathways. Materials and Methods: The PCOS model was created by injecting estradiol valerate into 60 adult Wistar rats, which were assigned to six groups of 10 animals: normal, PCOS, metformin, RCSO, and co-treatment with MET and RCSO groups. After 50 days, the serum levels of testosterone, progesterone, LH, and FSH hormones were measured, along with the activity of catalase, superoxide dismutase, glutathione peroxidase, and nitric oxide. Additionally, ovarian tissues were taken for gene expression analysis of Bax/Bcl-2/p53 and Cas-3 using real-time PCR, as well as protein expression analysis with western blot. Immunohistochemistry was performed to assess the levels of p53 and Ki-67 proteins, indicating the apoptosis of ovarian granulosa cells. Results: RCSO, in synergy with MET, demonstrated reductions in the LH/FSH ratio along with a decrease in progesterone and increased activity of antioxidant enzymes. Additionally, RCSO, along with MET, upregulated the Bax/Bcl-2/p53 and Cas-3 pathways, resulting in increased p53 expression and decreased Ki-67-positive cells, indicative of enhanced apoptosis. Conclusions: These findings suggest the potential use of RCSO in women with ovulation disorders, particularly those with PCOS.

Platelet rich plasma alleviates endometritis induced by lipopolysaccharide in mice via inhibiting TLR4/NF-κB signaling pathway.

Endometritis is an inflammatory reaction of the lining of uterus, leading to the occurrence of infertility. Platelet rich plasma (PRP) has been proven to exhibit extremely effective for the treatment of endometrium-associated infertility, but the mechanism of its prevention for endometritis remains unclear. The present study aimed to investigate the protective effect of PRP against endometritis induced by lipopolysaccharide (LPS) and elucidate the mechanism underlying these effects. Mouse model of endometritis was established by intrauterine perfusion of LPS. PRP intrauterine infusion was administered at 24h after LPS induction. After another 24h, the uterine tissues were harvested to observe histopathological changes, production of proinflammatory cytokines, variation of the Toll-like receptor 4/nuclear factor κB (TLR4/NF-κB) signaling pathways, and validated the anti-inflammatory effect of PRP. The myeloperoxidase (MPO) activity and concentration of nitric oxide (NO) were determined using assay kit. Proinflammatory chemokines (tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6)) were measured by ELISA and Real-Time PCR. The activity of TLR4/NF-κB pathway in uterine tissues was measured by Western blotting. Hematoxylin-eosin staining (H&E) appeared that PRP remarkably relieved the impairment of uterine tissues. Detection of MPO activity and concentration of NO revealed that PRP treatment distinctly mitigated infiltration of inflammatory cells in mice with endometritis induced by LPS. PRP treatment significantly affected the expression of TNF-α, IL-1β, and IL-6. PRP was also found to suppress LPS-induced activation of TLR4/NF-κB pathway. PRP effectively alleviates LPS-induced endometritis via restraining the signal pathway of TLR4/NF-κB. These findings provide a solid foundation for PRP as a potential therapeutic agent for endometritis.

Influential versatile potential of Entada rheedii and Myristica beddomei – an ethnobotanical approach

Aim of the study is to determine the anti-inflammatory, antidiabetic (α-amylase activity) and radical scavenging potentials of both leaf and stem ethanolic extracts of Entada rheedii and Myristica beddomei. DPPH, ABTS, H2O2, superoxide, hydroxyl and nitric oxide activity relate to inhibition ratio with different concentrations of extracts (μg/mL). Antidiabetic (α-amylase activity) and anti-inflammatory activities were also analyzed which showed high radical scavenging activity in a dose depending manner. Comparatively, leaves and stems of Entada rheedii showed higher radical scavenging activities compared to Myristica beddomei. Furthermore, both leaves and stem of Entada rheedii showed appreciable α-amylase inhibitory effects when compared with acarbose (standard drug) and also possess a strong anti-inflammatory effect (Diclofenac sodium, standard drug), compared to Myristica beddomei extracts. Overall, our results established the key evidence for Entada rheedii and Myristica beddomei extracts to be considered as a natural therapeutic aliment as the substitution for oxidative stress.

Performance of nitric oxide in sepsis: a scoping review

Performance of nitric oxide in sepsis: a scoping review

POSTTRANSLATIONAL MODIFICATIONS OF THE SULFHYDRYL GROUP OF THE CYSTEINE RESIDUE OF GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE

This review considers the main types of oxidative posttranslational modi cations of the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) targeting the sulfhydryl group of the catalytic cysteine residue Cys152. The highly reactive sulfhydryl group of Cys152 in the active centre of GAPDH undergoes oxidation and S-nitrosylation, leading to enzyme inactivation and destabilization. Upon reversible oxidation of the sulfhydryl group to form cysteine-sulfenic acid, the enzyme loses dehydrogenase activity, but gains the ability to catalyze the acyl-phosphatase reaction. Hydrolysis of the product of the dehydrogenase reaction, 1,3-diphosphoglycerate, under the action of the oxidized GAPDH leads to uncoupling of oxidation and phosphorylation at this stage of glycolysis. The action of nitric oxide results in S-nitrosylation of Cys152 GAPDH and the subsequent formation of cysteine-sulfenic acid due to hydrolysis of the S-NO-group. Data are presented on the relationship between S-nitrosylation of the catalytic Cys152 of GAPDH and its oxidation followed by S-glutathionylation of the enzyme at Cys152. The role of posttranslational modi cations of the sulfhydryl group of the catalytic cysteine residue in the regulation of enzyme activity, as well as the mechanisms ensuring the reversibility of such modi cations are discussed.

Melanoidin-like carbohydrate-containing macromolecules from Shanxi aged vinegar exert immunoenhancing effects on macrophage RAW264.7 cells

Bioactive macromolecule mining is important for the functional chemome analysis of traditional Chinese vinegar. In this study, we isolated and characterized carbohydrate-containing macromolecules from Shanxi aged vinegar (CCMSAV) and evaluated their immunomodulatory activity. The isolation process involved ethanol precipitation, deproteinization, decolorization, and DEAE-650 M column chromatography, resulting in the acquisition of four sub-fractions. All sub-fractions exhibited a molecular weight range of 6.92 to 16.71 kDa and were composed of 10 types of monosaccharides. Comparative analysis of these sub-fractions with two melanoidins exhibited similarities in elemental composition, spectral signature, and pyrolytic characteristics. Immunological assays confirmed the significantly enhanced cell viability, phagocytic activity, and secretion of nitric oxide, tumor necrosis factor (TNF)-α and interleukin (IL)-6 in RAW264.7 cells by all four sub-fractions. Further investigation of the immunomodulatory mechanism revealed that SAV-RP70-X, the most potent purified sub-fraction, enhanced aerobic glycolysis in macrophages and activated Toll-like receptor 2 (TLR2), TLR4, mannose receptor (MR), scavenger receptor (SR), and the dendritic cell-associated C-type lectin-1 receptor (Dectin-1). Furthermore, the activation of macrophages was associated with the MyD88/PI3K/Akt/NF-κB signaling pathway. Methylation analysis revealed that 1,4-Xylp was the most abundant glycosidic linkage in SAV-RP70-X.

Application of Bacillus Coagulans as Paraprobiotic Against Acute Hepatopancreatic Necrosis Disease in Shrimp.

Paraprobiotics, known as non-viable or ghost probiotics, have attracted attention for their benefits over live microbial cells. This study was designed to investigate the paraprobiotic effects of heat-killed Bacillus coagulans on the white leg shrimp Litopenaeus vannamei. The paraprobiotic formulation was prepared in three different concentrations including B. coagulans 1 (107 cells g-1 diet), B. coagulans 2 (108 cells g-1 diet), and B. coagulans 3 (109 cells g-1 diet) through heat inactivation method. Preliminary toxicity assessments revealed that post-larvae shrimps (mean weight ± SE: 0.025 ± 0.007g) treated with B. coagulans 1, 2 and 3 paraprobiotic formulations exhibited no mortality, confirming the non-toxic nature of the formulated diet. In a 90-day feeding trial involving juvenile shrimps (mean weight ± SE: 0.64 ± 0.05g), growth parameters and feed conversion ratios improved in all experimental groups. Subsequently, these shrimps were challenged with Vibrio parahaemolyticus, revealing that paraprobiotic-fed shrimps exhibited significant survival rate improvements. Oxidative stress-related enzyme activities, such as superoxide dismutase and catalase, increased in paraprobiotic-fed shrimps post-Vibrio challenge, while the challenged control group showed decreased activity (p < 0.001). Nitric oxide levels are also increased in paraprobiotic-treated shrimp, with B. coagulans 3 showing a significant rise in nitric oxide activity (p < 0.001). This study further demonstrated the positive impact of paraprobiotic treatment on digestive enzymes, immune-related parameters (e.g., total hemocyte count, prophenoloxidase, and respiratory burst activity), and overall disease resistance. These findings suggest that B. coagulans paraprobiotics have the potential to enhance antioxidant, antibacterial, and immune-related responses in L. vannamei, making them a valuable addition to shrimp aquaculture.

Effects of extended dietary supplementation with Santalum album essential oil on hemato-biochemical changes, innate immune response, antioxidant status, and expression of related gene in Nile tilapia (Oreochromis niloticus).

The effects of long-term dietary supplementation with sandalwood (Santalum album L.) essential oil (SEO) was investigated on hemato-biochemical biomarkers, immune status, antioxidant capacity, and resistance against Staphylococcus aureus in Nile tilapia (Oreochromis niloticus). Five groups (with four replicates) of O. niloticus (12.60 ± 0.20 g) were fed diets supplemented with SEO at doses of 0, 0.5, 1.0, 2.0, and 4.0 mL/kg diet for 60 days. Results indicated a substantial increase in blood protein levels and lower serum cholesterol, cortisol, glucose, urea, creatinine levels and, transaminase activities of fish fed a 2.0-mL SEO/kg diet. Serum lysozyme activity, nitric oxide, complement-3 levels, and phagocytic activity were significantly improved in O. niloticus after 60 days of feeding SEO-supplemented diets. Dietary SEO at level of 2.0-mL SEO/kg diet increased the activities of SOD, CAT, and GPx, and decreased MDA levels in liver homogenate. In addition, dietary 2.0-mL SEO/kg diet significantly upregulated antioxidant genes expression (CAT, SOD, GPx, GST, and GSR) with downregulation of apoptotic genes (HSP70, TLR2, caspase-3, and PCNA) in the liver. Furthermore, SEO-enriched diets significantly down-regulated pro-inflammatory (TNF-α, IL-1β, and IL-8) and up-regulated anti-inflammatory cytokine genes (TFG-β and IL-10) in the spleen. Moreover, SEO fortification increased the relative percentage of survival against S. aureus challenge and regulated immune-antioxidant genes in the spleen after the challenge. Overall, the results revealed that long-term using SEO might strengthen the physiological performance, hepatic oxidant/antioxidant balance, innate immune response, and resistance of O. niloticus against bacterial infections.

Berberine alleviates chlorpyrifos-induced nephrotoxicity in rats via modulation of Nrf2/HO-1 axis

Chlorpyrifos (CPS), an organophosphorus insecticide, is widely used for agricultural and non-agricultural purposes with hazardous health effects. Berberine (BBR) is a traditional Chinese medicine and a phytochemical with anti-inflammatory and anti-oxidative properties. The present study evaluated the effects of BBR against kidney damage induced by CPS and the underlying mechanisms. An initial study indicated that BBR 50 mg/kg was optimal under our experimental conditions. Then, 24 rats (6/group) were randomized into: control, BBR (50 mg/kg/day), CPS (10 mg/kg/day), and CPS + BBR. BBR was administration 1 h prior to CPS. Each treatment was delivered daily for a period of 28 consecutive days using a gastric gavage tube. Compared to CPS-alone treated rats, BBR effectively improved renal function by preventing the rise in serum urea, creatinine, and uric levels. The reno-protective effects of BBR were confirmed through a histological examination of kidney tissues. BBR restored oxidant-antioxidant balance in renal tissues mediated by Keap1/Nrf2/HO-1 axis modulation. In addition, BBR decreased nitric oxide (NO) and myeloperoxidase (MPO) activity. This was paralleled with the potent down-regulation of NF-κB. Furthermore, BBR exhibited anti-apoptotic activities supported by the upregulation of Bcl-2 and down-regulation of Bax and caspase-3 expression. In conclusion, our data suggest that BBR attenuates CPS-induced nephrotoxicity in rats by restoring oxidant-antioxidant balance and inhibiting inflammatory response and apoptosis in renal tissue. This is mediated, at least partly, by modulation of the Nrf2/HO-1 axis.

Anti-Inflammatory Activity of Vacuum Distillate from Panax ginseng Root on LPS-Induced RAW264.7 Cells.

Panax ginseng has been widely applied as an important herb in traditional medicine to treat numerous human disorders. However, the inflammatory regulation effect of P. ginseng distillate (GSD) has not yet been fully assessed. To determine whether GSD can ameliorate inflammatory processes, a GSD was prepared using the vacuum distillation process for the first time, and the regulation effect on lipopolysaccharide-induced macrophages was assessed. The results showed that GSD effectively inhibited nitric oxide (NO) formation and activation of inducible nitric oxide synthase (iNOS) mRNA in murine macrophage cell, but not cyclooxygenase-2 production. The mRNA expression pattern of tumor necrosis factor alpha and IL-6 were also reduced by GSD. Furthermore, we confirmed that GSD exerted its anti-inflammatory effects by downregulating c-Jun NH2-terminal kinase (JNK) phosphorylation, the extracellular signal-regulated kinase phosphorylation, and signaling pathway of nuclear factor kappa B (NF-κB). Our findings revealed that the inflammatory regulation activity of GSD could be induced by iNOS and NO formation inhibition mediated by regulation of nuclear factor kappa B and p38/JNK MAPK pathways.

In vivo and ex vivo studies support the immunostimulant and immunoprotective effect of Damiana (Turnera diffusa Willd) in Almaco Jack (Seriola rivoliana)

Damiana (Turnera diffusa Willd) was evaluated in vitro for antioxidant and antibacterial activities against Staphylococcus aureus and Streptococcus pyogenes (as a preliminary screening assessment) by high‐performance thin‐layer chromatography (HPTLC)-Direct bioautography. A study was performed in vivo to evaluate the effects of Damiana enriched diets at 0.5 % on immune parameters in mucus and serum and gene expression in Almaco Jack (Seriola rivoliana) intestine after two and four weeks; an infection with Aeromonas hydrophila at 1x107 colony forming units (CFU) followed and an ex vivo study was carried out using head-kidney leukocytes. Ferric reducing ability of plasma (FRAP) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays showed high antioxidant activities in Damiana leaves; even in the ABTS assay, Damiana at 300 μg/mL showed similar activity to ascorbic acid – the standard control. Damiana exhibited strong in vitro antimicrobial activity against S. aureus and S. pyogenes. In vivo studies showed a strong enhancement of myeloperoxidase, nitric oxide, superoxide dismutase, and catalase activities in mucus and serum of S. rivoliana supplemented with Damiana; their immunological response enhanced after infection with A. hydrophila. IL-1β, TNF-α, and IL-10 gene expressions upregulated in the fish intestine challenged with the bacterium. Piscidin and macrophage (MARCO) receptor gene expression up-regulated at week 4 and down-regulated after infection. Intestinal histology results confirm that Damiana not cause inflammation or damage. Finally, the ex vivo study confirmed the immunostimulant and protective effects of Damiana through increased phagocytic, respiratory burst, myeloperoxidase activities and nitric oxide generation before and upon the bacterial encounter. These results support the idea that Damiana has the potential as an immunostimulant additive for diets in aquaculture by enhancing immune parameters and protecting Almaco Jack against A. hydrophila infections upon four weeks of supplementation.

Deleterious Effects of Caffeine Consumption on Reproductive Functions of Female Wistar Rats.

The deleterious effects of caffeine consumption on reproductive functions of female Wistar rats were investigated in this study. In this experimental study, 35 female Wistar rats (180-200g) were divided into 7 groups: Control, II-IV received oral caffeine (10, 20, and 40mg/kg/day respectively) for 21 days. V-VII received similar caffeine doses for 21 days, followed by a 21-day withdrawal period. The ovaries, fallopian tubes, and uteri were assessed for levels of malondialdehyde (MDA), nitric oxide (NO), reduced glutathione (GSH), superoxide dismutase (SOD), and catalase activity using spectrophotometry. Serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), and estradiol levels were measured by ELISA. Organ histology was performed using microscopy. Statistical analysis employed ANOVA with significance at p<0.05. Caffeine caused dose-dependent increases in MDA, NO, and catalase activity in the ovaries, fallopian tubes, and uteri which decreased upon withdrawal. GSH levels in the ovary and fallopian tubes decreased with caffeine intake but recovered during withdrawal. Caffeine reduced estradiol levels in a dose-dependent manner, its withdrawal led to reductions in serum LH at 20 and 40mg/kg/day and FSH at 40mg/kg/day. Histology revealed dose-dependent alterations in ovarian architecture with congested connective tissues. Caffeine caused sloughing of plicae in the muscularis of the fallopian tubes, degenerated epithelial layer in the uterus, and severe inflammation of the myometrial stroma cells that persisted during caffeine withdrawal. Caffeine consumption adversely impacted the female reproductive functions of rats, altering hormonal balance and organ structure which persisted even after caffeine withdrawal.

In vitro evaluation against Leishmania amazonensis and Leishmania chagasi of medicinal plant species of interest to the Unified Health System.

Leishmaniasis is a disease of public health relevance that demands new therapeutic alternatives due to the toxicity of conventional treatments. In this study, 27 plants of interest to the Unified Health System (SUS) were evaluated for cytotoxicity in macrophages, leishmanicidal activity and production of nitric oxide (NO). None of the species demonstrated cytotoxicity to macrophages (CC50 >100 μg/mL). Extracts from Chenopodium ambrosioides, Equisetum arvense, Maytenus ilicifolia showed greater efficacy in inducing the death of Leishmania amazonensis amastigotes with IC50 of 68.4, 82.3, 75.7 μg/mL, respectively. The species Cynara scolymus, Punica granatum and Passiflora alata were the most effective in inducing an increase in the indirect concentration of NO (41.31, 29.30 and 28.86 µM, respectively) in cultures of macrophages infected with L. amazonensis. Furthermore, Punica granatum was also the most effective species in inducing an increase in NO in macrophages infected by Leishmania chagasi (19.90 µM). The results obtained so far support the continuation of studies, with the possibility of developing safer and more effective treatments for leishmaniasis, using natural products. The identification of plants that stimulate the production of NO in macrophages infected by Leishmania opens doors for more detailed investigations of the mechanism of action of these natural products.