HPLC-HESI-MS/MS Analysis of Phenolic Compounds from Cynoglossum tubiflorus Leaf Extracts: An Assessment of Their Cytotoxic, Antioxidant, and Antibacterial Properties.

The current study aimed to investigate the chemical composition, antioxidant, antibacterial, and cytotoxic properties of three extracts (hexane, dichloromethane, and methanol) from Cynoglossum tubiflorus. The composition of the methanolic extract was elucidated using HPLC-HESI-MS/MS analysis. The antioxidant effect was examined using NO, DPPH, FRAP, and TAC assays. Antimicrobial activity was evaluated by broth microdilution using various bacterial strains such as S. aureus, S. epidermidis, P. aeruginosa, E. coli, and K. pneumoniae. Structural disruptions in Gram-positive bacteria were visualized using scanning electron microscopy (SEM). Cytotoxic effects were evaluated on human MRC-5 in culture according to the MTT assay. The outcomes suggest that methanol extract contained a high amount of phenolic compounds (254.35 ± 0.360 mg GAE/g DE and 211.59 ± 0.939 mg QE/g DE). By applying the HPLC-HESI-MS/MS analysis, 32 compounds were identified, including phenolic acids, flavonoids, lignans, and fatty acids. This extract showed strong antioxidant (IC50 = 0.043 ± 0.001 mg/mL) and antimicrobial (MIC = 156 µg/mL) activities. The SEM suggests that cells exhibited membrane distortions characterized by surface depressions and alterations in bacterial shape, including dents, when compared to untreated cells. The in vitro cytotoxicity effect on human MRC-5 cells showed no toxicity effects at a concentration of 600 µg/mL. In silico analysis predicted low toxicity for all tested compounds across four different administration routes. This research indicates that this plant could be explored as a powerful source of natural drugs to target pathogens, with applications in the food, pharmaceutical, and medical industries.

Vitamin B1 deficiency leads to high oxidative stress and mtDNA depletion caused by SLC19A3 mutation in consanguineous family with Leigh syndrome.

Leigh syndrome (LS) and Leigh-like spectrum are the most common infantile mitochondrial disorders characterized by heterogeneous neurologic and metabolic manifestations. Pathogenic variants in SLC carriers are frequently reported in LS given their important role in transporting various solutes across the blood-brain barrier. SLC19A3 (THTR2) is one of these carriers transporting vitamin-B1 (vitB1, thiamine) into the cell. Targeted NGS of nuclear genes involved in mitochondrial diseases was performed in a patient belonging to a consanguineous Tunisian family with LS and revealed a homozygous c.1264 A > G (p.T422A) variant in SLC19A3. Molecular docking revealed that the p.T422A aa change is located at a key position interacting with vitB1 and causes conformational changes compromising vitB1 import. We further disclosed decreased plasma antioxidant activities of CAT, SOD and GSH enzymes, and a 42% decrease of the mtDNA copy number in patient blood.Altogether, our results disclose that the c.1264 A > G (p.T422A) variant in SLC19A3 affects vitB1 transport, induces a mtDNA depletion and reduces the expression level of oxidative stress enzymes, altogether contributing to the LS phenotype of the patient.

The pyrethroid insecticide permethrin confers hepatotoxicity through DNA damage and mitochondria-associated apoptosis induction in rat: Palliative benefits of Fumaria officinalis.

Permethrin (PER) is a pyrethroid pesticide that is extensively used as an insecticide in world because of its high activity and its low mammalian toxicity. The current study was conducted to investigate the protective action of Fumaria officinalis against PER-induced liver injury in male rats. However, HPLC-DAD showed the richness of 6 components in F. officinalis (F) including quercetin, ferulic acid, and naringenin which were the most abundant. Total polyphenols, total flavonoids, and condensed tannins were studied by phytochemical screening. In vitro, antioxidant properties showed that F. officinalis exhibited the highest DPPH radical, FRAP, and H2 O2 tests and total antioxidant capacity. Wistar rats were divided into four groups: negative control group (C), positive control group (F) (200 mg F. officinalis/kg BW), PER group (34.05 mg permethrin/kg BW), and PER + F group (34.05 mg permethrin/kg BW and 200 mg F. officinalis/kg BW). Oral administration of PER led to promote a decrease of body weight and Ca2+ -ATPases and Mg2+ -ATPases activities and an increase of plasma C-reactive protein level, transaminases, and hepatic ϒ-GT activities as well as hepatic and mitochondrial oxidative stress. An increase in plasma lactate-to pyruvate ratio and a reduction in complexes enzymes I, III, and IV activities were also observed. In addition, histoarchitecture of liver in PER-treated rats showed apoptosis and necrosis as confirmed by DNA fragmentation. F. officinalis significantly exerted hepatoprotective effect by modulating hepatic alteration and mitochondrial dysfunction as well as genotoxicity. This effect could be attributed to phenolics compounds such as polyphenols, condensed tannins, and flavonoids.

Mitochondrial bioenergetics and redox dysfunction in nephrotoxicity induced by pyrethroid permethrin are ameliorated by flavonoid-rich fraction.

The present study was designed to evaluate in vitro and in vivo the potential anti-inflammatory and nephroprotective potential of ethyl acetate fraction extracted from Fumaria officinalis (EAF) against permethrin (PER). Male wistar rats were treated daily by gavage during 7 days as follows: group C: negative control rats received 2 mL/kg bw of corn oil, group EAF: positive control rats received EAF at a dose of 200 mg/kg bw dissolved in water, group PER: rats received PER at a dose of 34.05 mg/kg bw and group (PER + EAF): rats received PER (34.05 mg/kg bw) and EAF (200 mg/kg bw). In vitro study showed the ability of EAF to inhibit protein denaturation and heat-induced hemolysis confirming its anti-inflammatory activity. In vivo, PER treatment decreased calcium (Ca) and phosphorus (P) levels and increased lactate dehydrogenase (LDH) activity in plasma. It induced oxidative stress objectified by an increase in the lipid peroxidation and protein oxidation and a perturbation of antioxidant system in kidney and mitochondria. The activities of NADH-ubiquinone reductase, ubiquinol-cytochrome C reductase and cytochrome C oxidase activities were reduced. These alterations were confirmed by histopathological studies. Co-treatment with EAF improved the antioxidant status and mitochondrial bioenergetics. The nephroprotective effects of EAF could be attributed to its modulation of detoxification enzymes and/or free radical scavenging actions.

Subacute toxicity studies of meals prepared from genetically modified potato overexpressing the StDREB1 or the VvWRKY2 transcription factor in rats.

BACKGROUND: Potato tubers from genetically modified plants overexpressing the StDREB1 or the VvWRKY2 transcription factors that exhibited improved tolerance to salt and resistance to Fusarium solani infection were characterized and evaluated for safety in a 30 day rat feeding study. Male Wistar rats were split into four groups and provided with a diet composed of 33% (w/w) of either one of the two genetically modified potatoes (GMPs), 33% of the commercial Spunta variety (Sp), or a control group fed with the basal rats' diet. The influence of the GMPs on rat behavior and overall health parameters was evaluated and compared with that of commercial potato (i.e. the Sp group) and control diet. RESULTS: Small differences were noticed in the chemical composition of the different tubers, but all the diets were adjusted to an identical caloric level. Results showed no sign of toxic or detrimental effects on the rats' overall health as a result of these diets. The rats fed with the GMPs meal showed hematological and biochemical compositions of the plasma comparable to the control groups. No histopathological damage nor any structural disorganization, severe congestion, or acute inflammation were noticed in the rats' tissues. CONCLUSION: Under these study conditions, the GMP diets did not induce any apparent or significant adverse effects on rats after 30 days of dietary administration in comparison with rats fed diets with the corresponding non-transgenic diet and the standard diet group. These two GMPs were therefore considered to be as safe as their commercial comparator. © 2022 Society of Chemical Industry.

Effect of Medicago sativa compared To 17ß-oestradiol on osteoporosis in ovariectomized mice.

Phytoestrogens, with a wide range of beneficial effects, prevent bone loss caused by oestrogen deficiency.The purpose of this study was to evaluate the effect of Medicago sativa ethanol extract compared to 17ß-oestradiol on osteoporosis in ovariectomized mice.The study was carried out on female mice, divided into five groups: control mice (GI), Medicago sativa treated mice (0.75 g/kg BW/day) (GII), ovariectomized mice (GIII) and ovariectomized mice treated either with Medicago sativa (GIV) or with 17ß-oestradiol (50 µg/Kg BW/day) (GV).Our results showed that Medicago sativa or 17ß-oestradiol treatments significantly attenuated perturbations of mineral levels, histological changes and oxidative stress in the femurs of ovariectomized mice.Medicago sativa prevented bone loss induced by oestrogen deficiency, which could be attributed to its richness in kaempferol, syringic acid, naringenin and myrictin. Its effects were more beneficial or similar compared to 17ß-oestradiol.

Flavonoid-rich fraction attenuates permethrin-induced toxicity by modulating ROS-mediated hepatic oxidative stress and mitochondrial dysfunction ex vivo and in vivo in rat.

The present study explores the antioxidant, anti-microbial, and hepatoprotective potentials of flavonoid-rich fractions from Fumaria officinalis against permethrin-induced liver damage ex vivo/in vivo in rat. However, HPLC-DAD analysis revealed the richness of 6 components in ethyl acetate fraction (EAF) where ferulic acid, rosmarinic acid, and myricetin are the most abundant. The in vitro assays showed that EAFs have impressive antioxidant and anti-microbial properties. Ex vivo, permethrin (PER) (100 µM) induced a decrease of hepatic AST and ALT activities and 25-OH vitamin D and vitamin C levels and an increase of ALP and LDH activities, TBARS, and ϒ-GT levels with a disturbance of oxidative status. The hepatoprotective effect of EAF (1 mg/mL) against PER was confirmed by the amelioration of oxidative stress profile. In vivo, permethrin was found to increase absolute and relative liver weights, plasma transaminase activities, lactate-to-pyruvate ratio, hepatic and mitochondrial lipid peroxidation, and protein oxidation levels. This pesticide triggered a decrease of Ca2+ and Mg2+-ATPases and mitochondrial enzyme activities. The co-treatment with EAF reestablished the hepatic and mitochondrial function, which could be attributed to its richness in phenolic compounds.

Effects of estrogen deficiency on liver function and uterine development: assessments of Medicago sativa's activities as estrogenic, anti-lipidemic, and antioxidant agents using an ovariectomized mouse model.

We investigated the effects of Medicago sativa supplementation on the lipid profiles and antioxidant capacities of ovariectomized mice.The study was performed on white Swiss female mice that were divided into five groups: control, treated with Medicago sativa (0.75 g/kg/day), ovariectomized, ovariectomized treated with ß-estradiol (1 µg/day) or with Medicago sativa. The mice were sacrificed after 3 and 8 weeks of treatment.Ovariectomy induced a decrease in overall growth, uterine atrophy, and hyperlipidemia demonstrated by increased cholesterol, triglycerides, and decreased HDL. We have shown the involvement of oxidative stress in this hepatic lesion proven by increased levels of TBARS, GPX, and GSH, and decreased levels of SOD and catalase.Treatment with Medicago sativa restores lipid balance, the activity of antioxidant enzymes and improves lipid peroxidation. This is probably due to the richness of this plant in polyphenols and flavonoids considered as an antioxidant and phytoestrogenic elements.

Polysaccharides from Phormidium versicolor (NCC466) protecting HepG2 human hepatocellular carcinoma cells and rat liver tissues from cadmium toxicity: Evidence from in vitro and in vivo tests.

The in vitro antioxidant, cytotoxic and cytoprotective properties and in vivo hepatoprotective activities of crude polysaccharides extracted from cyanobacteria Phormidim versicolor NCC466 (CFv-PS) were investigated. The CFv-PS, identified as heteropolysaccharides with molecular weight of 63.79kDa, exhibited relatively strong antioxidant activity, in a concentration-depended manner, in vitro assays. Additionally, CFv-PS did not induce cytotoxic effect on HepG2 human hepatocellular carcinoma cells within the range of tested concentrations (25-150µg·mL-1) while preventing them against Cd. Moreover, in rats subjected to Cd-induced hepatotoxicity, CFv-PS pretreatment significantly (P<0.05) reduced the level of ALAT, ASAT, biliburin, MDA, protein carbonyl and DNA damage, and markedly increased enzyme activities in liver tissues. These findings suggest that the cyanobacteria Phormidium versicolor is a potential source of natural products possessing antioxidant, cytoprotective and hepatoprotective properties.