NARINGIN AND RUTIN PREVENT D-GALACTOSAMINE-INDUCED HEPATIC INJURY IN RATS VIA ATTENUATION OF THE INFLAMMATORY CASCADE AND OXIDATIVE STRESS

The dried stems and leaves of Citrus jambhiri Lush. (Rutaceae) were extracted with aqueous methanol and the extracts were fractionated using light petroleum, chloroform and ethyl acetate. Column chromatography of the ethyl acetate fractions resulted in the isolation of naringin, rutin, hesperidin, and neohesperidin. Their structures were identified by MS and different NMR techniques. Liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) of these fractions allowed the identification 7 flavonoid glycosides. Hepatoprotective properties of naringin and rutin were evaluated in d-galactosamine (d-GalN)-induced hepatic injury in rats. d-GalN increased serum aminotransferase activity, total bilirubin, liver tumor necrosis factor-α level (TNF-α), hepatic lipid peroxidation, nuclear factor κB and decreased hepatic glutathione content, IL-10 levels and the IL-10/TNF-α ratio. These changes were attenuated in rats pretreated with rutin and naringin (40 mg/kg body weight). They increased liver IL-10 levels and the IL-10/TNF-α ratio. Rutin but not naringin down-regulated NF-κB gene expression and decreased gamma-glutamyltransferase (GGT) activity

In vitro cytotoxicity of Withania somnifera (L.) roots and fruits on oral squamous cell carcinoma cell lines: a study supported by flow cytometry, spectral, and computational investigations

Oral cancer is a severe health problem that accounts for an alarmingly high number of fatalities worldwide. Withania somnifera (L.) Dunal has been extensively studied against various tumor cell lines from different body organs, rarely from the oral cavity. We thus investigated the cytotoxicity of W. somnifera fruits (W-F) and roots (W-R) hydromethanolic extracts and their chromatographic fractions against oral squamous cell carcinoma (OSCC) cell lines [Ca9-22 (derived from gingiva), HSC-2, HSC-3, and HSC-4 (derived from tongue)] and three normal oral mesenchymal cells [human gingival fibroblast (HGF), human periodontal ligament fibroblast (HPLF), and human pulp cells (HPC)] in comparison to standard drugs. The root polar ethyl acetate (W-R EtOAc) and butanol (W-R BuOH) fractions exhibited the strongest cytotoxicity against the Ca9-22 cell line (CC50 = 51.8 and 40.1 μg/mL, respectively), which is relatively the same effect as 5-FU at CC50 = 69.4 μM and melphalan at CC50 = 36.3 μM on the same cancer cell line. Flow cytometric analysis revealed changes in morphology as well as in the cell cycle profile of the W-R EtOAc and W-R BuOH-treated oral cancer Ca9-22 cells compared to the untreated control. The W-R EtOAc (125 μg/mL) exerted morphological changes and induced subG1 accumulation, suggesting apoptotic cell death. A UHPLC MS/MS analysis of the extract enabled the identification of 26 compounds, mainly alkaloids, withanolides, withanosides, and flavonoids. Pharmacophore-based inverse virtual screening proposed that BRD3 and CDK2 are the cancer-relevant targets for the annotated withanolides D (18) and O (12), and the flavonoid kaempferol (11). Molecular modeling studies highlighted the BRD3 and CDK2 as the most probable oncogenic targets of anticancer activity of these molecules. These findings highlight W. somnifera’s potential as an affordable source of therapeutic agents for a range of oral malignancies

Phenolic Compounds from <i>Populus alba</i> L. and <i>Salix subserrata</i> Willd. (Salicaceae) Counteract Oxidative Stress in <i>Caenorhabditis elegans</i>

Utilizing bioassay- and TLC-guided column chromatography, fifteen secondary metabolites from Populus alba and eight compounds from Salix subserrata were isolated, including a novel plant metabolite salicyl ether and characterized using ultralviolet light (UV) absorbance, mass spectrometry (MS), 1H-, 13C-NMR (nuclear magnetic resonance), heteronuclear single quantum coherence spectroscopy (HSQC) and heteronuclear multiple bond correlation (HMBC). The extracts, their sub-fractions and the isolated compounds exhibited promising antioxidant activities in vitro in DPPH and FRAP assays. Also, the extracts of P. alba leaf (PL), shoots (PS), and S. subserrata leaf (SL) demonstrated substantial antioxidant activities in vivo in the multicellular model organism Caenorhabditis elegans. For the first time, the isolated secondary metabolites, aromadendrin, tremuloidin, salicin, isorhamnetin-3-O-&#946;-d-rutinoside, gallocatechin, triandrin, and chrysoeriol-7-O-glucuronide were investigated. They exhibited substantial antioxidant activities in vivo. Salicin, isorhamnetin-3-O-&#946;-d-rutinoside and gallocatechin, in particular, protected the worms against a lethal dose of the pro-oxidant juglone (80 &#181;M), decreased the endogenous reactive oxygen species (ROS) level to 45.34%, 47.31%, 68.09% and reduced juglone- induced hsp-16.2::GFP (green fluorescence protein) expression to 79.62%, 70.17%, 26.77%, respectively. However, only gallocatechin induced higher levels of sod-3 expression. These findings support the traditional use of Populus alba and Salix subserrata for treating inflammation especially when ROS are involved

Carissa macrocarpa Leaves Polar Fraction Ameliorates Doxorubicin-Induced Neurotoxicity in Rats via Downregulating the Oxidative Stress and Inflammatory Markers

Chemotherapeutic-related toxicity exacerbates the increasing death rate among cancer patients, necessitating greater efforts to find a speedy solution. An in vivo assessment of the protective effect of the C. macrocarpa leaves polar fraction of hydromethanolic extract against doxorubicin (Dox)-induced neurotoxicity was performed. Intriguingly, this fraction ameliorated Dox-induced cognitive dysfunction; reduced serum ROS and brain TNF-&alpha; levels, upregulated the brain nerve growth factor (NGF) levels, markedly reduced caspase-3 immunoexpression, and restored the histological architecture of the brain hippocampus. The in vivo study results were corroborated with a UPLC-ESI-MS/MS profiling that revealed the presence of a high percentage of the plant polyphenolics. Molecular modeling of several identified molecules in this fraction demonstrated a strong binding affinity of flavan-3-ol derivatives with TACE enzymes, in agreement with the experimental in vivo neuroprotective activity. In conclusion, the C. macrocarpa leaves polar fraction possesses neuroprotective activity that could have a promising role in ameliorating chemotherapeutic-induced side effects

Chemical profiling and cytotoxic potential of the n-butanol fraction of Tamarix nilotica flowers

Abstract Background Cancer represents one of the biggest healthcare issues confronting humans and one of the big challenges for scientists in trials to dig into our nature for new remedies or to develop old ones with fewer side effects. Halophytes are widely distributed worldwide in areas of harsh conditions in dunes, and inland deserts, where, to cope with those conditions they synthesize important secondary metabolites highly valued in the medical field. Several Tamarix species are halophytic including T.nilotica which is native to Egypt, with a long history in its tradition, found in its papyri and in folk medicine to treat various ailments. Methods LC–LTQ–MS–MS analysis and 1H-NMR were used to identify the main phytoconstituents in the n- butanol fraction of T.nilotica flowers. The extract was tested  in vitro for its cytotoxic effect against breast (MCF-7) and liver cell carcinoma (Huh-7) using SRB assay. Results T.nilotica n-butanol fraction of the flowers was found to be rich in phenolic content, where, LC–LTQ–MS–MS allowed the tentative identification of thirty-nine metabolites, based on the exact mass, the observed spectra fragmentation patterns, and the literature data, varying between tannins, phenolic acids, and flavonoids. 1H-NMR confirmed the classes tentatively identified. The in-vitro evaluation of the n-butanol fraction showed lower activity on MCF-7 cell lines with IC50 > 100 µg/mL, while the higher promising effect was against Huh-7 cell lines with an IC50= 37 µg/mL. Conclusion Our study suggested that T.nilotica flowers' n-butanol fraction is representing a promising cytotoxic candidate against liver cell carcinoma having potential phytoconstituents with variable targets and signaling pathways

Human Lung Cancer (A549) Cell Line Cytotoxicity and Anti-Leishmania major Activity of Carissa macrocarpa Leaves: A Study Supported by UPLC-ESI-MS/MS Metabolites Profiling and Molecular Docking

Lung cancer and cutaneous leishmaniasis are critical diseases with a relatively higher incidence in developing countries. In this research, the activity of Carissa macrocarpa leaf hydromethanolic extract and its solvent-fractions (n-hexane, EtOAc, n-butanol, and MeOH) against the lung adenocarcinoma cell line (A549) and Leishmania major was investigated. The MeOH fraction exhibited higher cytotoxic activity (IC50 1.57 &plusmn; 0.04 &mu;g/mL) than the standard drug, etoposide (IC50 50.8 &plusmn; 3.16 &mu;g/mL). The anti-L. major results revealed strong growth inhibitory effects of the EtOAc fraction against L. major promastigotes (IC50 27.52 &plusmn; 0.7 &mu;g/mL) and axenic amastigotes (29.33 &plusmn; 4.86% growth inhibition at 100 &mu;g/mL), while the butanol fraction exerted moderate activity against promastigotes (IC50 73.17 &plusmn; 1.62), as compared with miltefosine against promastigotes (IC50 6.39 &plusmn; 0.29 &mu;g/mL) and sodium stibogluconate against axenic amastigotes (IC50 22.45 &plusmn; 2.22 &mu;g/mL). A total of 102 compounds were tentatively identified using UPLC-ESI-MS/MS analysis of the total extract and its fractions. The MeOH fraction was found to contain several flavonoids and flavan-3-ol derivatives with known cytotoxic properties, whereas the EtOAc fractions contained triterpene, hydroxycinnamoyl, sterol, and flavanol derivatives with known antileishmanial activity. Molecular docking of various polyphenolics of the MeOH fraction with HDAC6 and PDK3 enzymes demonstrates high binding affinity of the epicatechin 3-O-&beta;-D-glucopyranoside and catechin-7-O-&beta;-D-glucopyranoside toward HDAC6, and procyanidin C2, procyanidin B5 toward PDK3. These results are promising and encourage the pursuit of preclinical research using C. macrocarpa&rsquo;s MeOH fraction as anti-lung cancer and the EtOAc fraction as an anti-L. major drug candidates

Preparation and in vivo evaluation of nano sized cubosomal dispersion loaded with Ruta graveolens extracts as a novel approach to reduce asthma-mediated lung inflammation

Asthma is a chronic disease affecting people of all ages. Asthma medications are associated with adverse effects restricting their long-term usage, demanding newer alternative therapies. This study aimed to investigate the anti-asthmatic properties of Ruta graveolens extract and its prepared nano-cubosomal dispersion (Ruta-ND). Firstly, the R. graveolens methanolic extract exhibited higher anti-inflammatory activity on Lipopolysaccharide (LPS)-activated BEAS-2B cells. To ensure best bioavailability and hence best cellular uptake, R. graveolens extract was loaded in nano-cubosomal dispersion (ND). Then, the anti-asthmatic effects of Ruta extract and ND were simultaneously evaluated in rats’ model with ovalbumin-induced allergic asthma. R. graveolens extract and Ruta-ND subsided asthma score and improved lung function by restoring FEV1/FVC ratio to the expected values in control rats. Also, it showed strong antioxidant and anti-inflammatory activities manifested by lowering levels of malondialdehyde (MDA), IL-4, IL-7, TGF-β, and Ig-E, and increasing levels of superoxide dismutase (SOD) and INF-γ in bronchoalveolar lavage fluid. Our research findings also indicate autophagy induction and apoptosis inhibition by Ruta extract and Ruta-ND. Finally, the HPLC MS/MS phytochemical profiling of R. graveolens extract evident production of various alkaloids, flavonoids, coumarins, and other phenolics with reported pharmacological properties corresponding to/emphasize our study findings. In conclusion, R. graveolens exhibited promise in managing Ova-induced allergic asthma and could be developed as an alternative anti-allergic asthma drug