Punicalagin Protects against the Development of Methotrexate-Induced Hepatotoxicity in Mice via Activating Nrf2 Signaling and Decreasing Oxidative Stress, Inflammation, and Cell Death.

Despite its effectiveness in treating inflammatory diseases and various malignancies, methotrexate (MTX) is well known to cause hepatotoxicity, which involves increased oxidative stress and inflammation, limiting its clinical use. Herein, we looked into the effect of punicalagin (PU), a polyphenolic molecule having a variety of health-promoting attributes, on MTX-induced hepatotoxicity in mice. PU (25 and 50 mg/kg/day) was given orally to the mice for 10 days, while a single dose of MTX (20 mg/kg) was injected intraperitoneally (i.p.) at day 7. The MTX-induced liver damage was demonstrated by remarkably higher transaminases (ALT and AST), ALP, and LDH, as well as significant histological alterations in hepatic tissues. MTX-injected mice also demonstrated increases in hepatic oxidative stress markers, including malondialdehyde (MDA) and nitric oxide (NO), with a concordant drop in glutathione (GSH) content and superoxide dismutase (SOD) and catalase (CAT) activities. PU significantly attenuated the MTX-induced serum transaminases, ALP and LDH elevations, and hepatic oxidative stress measures and boosted antioxidant defenses in the liver. Moreover, the liver of MTX-treated mice showed increases in NF-κB p65 expression, pro-inflammatory cytokine (IL-6 and TNF-α) levels, and pro-apoptotic protein (caspase-3 and Bax) expression, whereas Bcl-2 and Nrf2 expressions were reduced, which were all attenuated by PU treatment. Collectively, PU inhibits oxidative damage, inflammation, and apoptosis and upregulates Nrf2 in the liver of MTX-induced mice. Thus, these findings suggest that PU may have great therapeutic potential for the prevention of MTX-induced hepatotoxicity, pending further exploration in upcoming studies.

Punicalagin attenuates myocardial oxidative damage, inflammation, and apoptosis in isoproterenol-induced myocardial infarction in rats: Biochemical, immunohistochemical, and in silico molecular docking studies.

Myocardial infarction (MI) is a life-threatening ischemic disease and is one of the leading causes of morbidity and mortality worldwide. Punicalagin (PU), the major ellagitannin found in pomegranates, is characterized by multiple antioxidant activities. The aim of this study is to assess the protective effects of PU against isoproterenol (ISO)-induced acute myocardial damage and to investigate its underlying vascular mechanisms using rat model. METHODS: Rats were randomly divided into five groups and were treated orally (p.o.) with PU (25 and 50 mg/kg) for 14 days. ISO was administered subcutaneously (S.C.) (85 mg/kg) on the 15th and 16th days to induce Myocardial infarction. Cardiac markers, oxidative stress markers, and inflammatory cytokines levels were determined in the heart tissue. Immunohistochemistry analysis was performed to determine the protein expression pathways of inflammation, apoptosis and oxidative stress (Nuclear factor erythroid 2-related factor 2 (Nrf-2), and heme oxygenase-1 (HO-1) in all the groups. In silico study was carried out to evaluate the molecular interaction of PU with some molecular targets. RESULTS: Our results showed that ISO-induced cardiac tissue injury was evidenced by increased serum creatine kinase-MB (CK-MB), cardiac troponin I (cTnI), and lactate dehydrogenase (LDH), associated with several histopathological changes. ISO also induced an increase of MDA, PCO, NO, and 8-hydroxy-2-deoxyguanosine (8-OHdG), along with a decrease of antioxidant enzyme activities in the myocardial tissues. In addition, an increase of TNF-α, NF-κB, IL-6, IL-1ß, iNOS, Nrf2 and (HO-1) was observed. Pre-treatment with PU reduced myocardial infract area, ameliorated histopathological alterations in myocardium, and decreased activities of myocardial injury marker enzymes in ISO-induced rats. In addition, PU remarkably restored ISO-induced elevation of lipid peroxidation and decrease of antioxidants, significantly reduced myocardial pro-inflammatory cytokines concentrations in this animal model. Molecular docking analysis of PU with protein targets showed potent interactions with negative binding energies. In conclusion, PU can protect the myocardium from oxidative injury, inflammatory response, and cell death induced by ISO by upregulating Nrf2/HO-1 signaling and antioxidants.

Hepatoprotective effect of taxifolin on cyclophosphamide-induced oxidative stress, inflammation, and apoptosis in mice: Involvement of Nrf2/HO-1 signaling.

Taxifolin (TA) is a natural flavonoid found in many foods and medicinal plants with well-documented antioxidant and anti-inflammatory properties. Cyclophosphamide (CP) is an effective antineoplastic and immunosuppressive agent; however, it is associated with numerous adverse events, including hepatotoxicity. Herein, we aimed to investigate the potential protective effects of TA using a mouse model of CP-induced hepatotoxicity. Mice were co-treated with TA (25 and 50 mg/kg, orally) and CP (30 mg/kg, i.p.) for 10 consecutive days and sacrificed 24 hours later. CP induced increased transaminases (ALT and AST), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) paralleled with pronounced histopathological alterations in the liver. Moreover, hepatic tissues of CP-injected mice showed increased malondialdehyde (MDA), protein carbonyl, and nitric oxide (NO) levels, accompanied by decreased antioxidant defenses (glutathione [GSH], superoxide dismutase [SOD], and catalase [CAT]). Livers of CP-injected mice also showed increased inflammatory response (nuclear transcription factor kappa-B [NF-κB] p65 activation, increased levels of proinflammatory cytokines tumor necrosis factor alpha [TNF-α], interleukin 1 beta [IL-1ß], and IL-6) and apoptosis (decreased Bcl-2 and increased Bax and caspase-3 expression levels). Remarkably, TA ameliorated markers of liver injury and histological damage in CP-injected mice. TA treatment also attenuated numerous markers of oxidative stress, inflammation, and apoptosis in the liver of CP-injected mice. This was accompanied by increased nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) expression in the liver tissues of CP-injected mice. Taken together, this study indicates that TA may represent a promising new avenue to prevent/treat CP-induced hepatotoxicity and perhaps other liver diseases associated with oxidative stress and inflammation.

Consequences of supplementing duck's diet with charcoal on carcass criteria, meat quality, nutritional composition, and bacterial load.

The influence of charcoal as feed additives on carcass and meat characteristics was studied in 144 four weeks old Muller ducks. The experimental ducklings were assigned to six groups of 24 birds (Eight per replicates each). The dietary treatments contained 0, 0.5, 1.0, 1.5, 2.0, and 2.5% charcoal for G1 (C), G2 (L1), G3 (L2), G4 (L3), G5 (L4) and G6 (L5), respectively. All experimental birds were raised under similar environmental and managerial conditions. Results indicated that charcoal did not affect most carcass traits significantly except for dressing percentage was higher (P < 0.05) in 1.5 and 2 % charcoal included ducks diets compared to control ducks. Charcoal supplementation significantly affected duck meat tenderness, juiciness and water holding capacity. Moreover, charcoal altered (P < 0.05) meat components such as crude protein, calcium components, desirable fatty acids, nutritional value and some bacterial counts. Thiobarbituric acid reactive substances reduced in birds fed charcoal at 1.5, 2, and 2.5%, with significant variation among treatments. No significant differences in the number of Escherichia coli and Staphylococcus aureus were detected among the ducks fed with charcoal and the control group. It could be concluded that charcoal could be included in ducks' diets at 1.5 and 2% with beneficial effects on carcass parameters.

Chemical Characterization of Clove, Basil and Peppermint Essential Oils; Evaluating Their Toxicity on the Development Stages of Two-Spotted Spider Mites Grown on Cucumber Leaves.

The two spotted spider mite (TSSM), Tetranychus urticae Koch, is a cosmopolitan mite. It rapidly reproduces and can develop resistance to chemical pesticides. This study aims to evaluate the toxicity and acaricidal activity of three essential oils from basil, clove, and peppermint against T. urticae reproduction, which is grown on three cucumber cultivars, Chief (SC 4145), Raian (CB898), and Toshka (SC 349), under laboratory conditions at 27 + 3 °C and 70 + 5% RH. GC-MS characterized the volatile oils of basil, clove, and peppermint. Methyl cinnamate, eugenol, and menthol were the main essential oils in basil, clove, and peppermint, respectively. The results indicated significant differences in the duration of development between T. urticae feeding on the three cucumber cultivars (p ≤ 0.05), including eggs, protonymph, and deutonymph time. The Toshka (SC 349) cultivar recorded the lowest developmental time. The longevity period exhibited the same trend with non-significant differences between Raian (CB898) and Toshka (SC 349). Moreover, the lethal concentration (LC50) and LC90 values in tested essential oils (EOs) showed that clove EOs were the most toxic. In contrast, basil and peppermint EOs were the least effective, and immature stages were more sensitive to EOs than adult stages. The infected Toshka (SC 349) discs treated with essential oils and abamectin under in vitro conditions indicated that clove oil is comparable to abamectin regarding its effect on the egg numbers (18.7 and 17.6 egg), immature development time, longevity, life span, and life cycle (20.6 and 20.8 days) of T. urticae. We conclude that the resistant cultivation of cucumber plants can be recommended in integrated pest management programs. The most effective of the tested oils, clove EOs, should be used as alternatives to pesticides to control T. urticae in the protected cultivation of cucumbers.