Evaluation of cytotoxicity and oxidative stress induced by alcoholic extract and oil of Lepidium Sativum seeds in human liver cell line HepG2

Since, the primary site of drug metabolism is the liver, that plays a major role in metabolism, digestion, detoxification, and elimination of substances from the body, the present studies were designed to investigate the possible adverse effect of alcoholic extract of seeds of Lepidium sativum (LSA) and Lepidium sativum seed oil (LSO) on HepG2 cells, a human liver cell line. LSA and LSO induced cell viability by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and neutral red uptake (NRU) assays. Morphological changes, lipid peroxidation, glutathione, catalase, and superoxide dismutase activities in HepG2 cells were studied. Cells were exposed to 25 to 1000 μg/ml of LSA and LSO for 24 h. The results show that LSA and LSO reduced cell viability, and altered the cellular morphology in dose dependent manner. Concentrations (100 to 1000 μg/ml) of LSA and LSO were found to be cytotoxic, whereas 50 μg/ml and lower concentrations did not cause any significant adverse effect in cell viability of HepG2 cells. LSA and LSO were also found to induce oxidative stress in dosedependent manner indicated by decrease in glutathione level, catalase activity, and SOD activity and an increase in lipid peroxidation. The results indicate that LSA and LSO induced oxidative stress mediated cytotoxicity in HepG2 cells.Keywords: Lepidium sativum, HepG2 cells, oxidative stress, cytoxicityAfrican Journal of Biotechnology Vol. 12(24), pp. 3854-386

A facile one-pot synthesis of novel 2,5-disubstituted-1,3,4-oxadiazoles under conventional and microwave conditions and evaluation of their in vitro antimicrobial activities

AbstractA rapid and efficient solvent-free synthesis of 2,5-disubstituted-1,3,4-oxadiazoles (3a–l) from fatty acid hydrazides (1a–f) under microwave irradiation is described. The structural elucidation of these compounds is based on their spectral data (IR, 1H NMR, 13C NMR and MS). All the newly synthesized compounds have been screened for their antibacterial and antifungal activities. The compounds 3f, 3j and 3l were found to be most potent anti-microbial agents

Isolation, biological evaluation and validated HPTLC-quantification of the marker constituent of the edible Saudi plant Sisymbrium irio L.

AbstractPhytochemical investigation and chromatographic purification of the n-hexane fraction of the aerial parts of the edible Saudi plant Sisymbrium irio led to the isolation of β-sitosterol (1), stigmasterol (2) and β-sitosterol-β-d-glucoside (3). The cytotoxic effects of the n-hexane, dichloromethane, ethyl acetate and n-butanol fractions were tested against three cancer cell lines viz., MCF-7, HCT-116 and HepG2, using the crystal violet staining (CVS) method, while the antibacterial activity against a number of pathogenic bacterial strains, was also estimated using the broth microdilution assay. The n-hexane fraction showed potent cytotoxic activities against all tested human cancer cell lines (IC50: 11.7–13.4μg/mL), while the dichloromethane fraction was particularly potent against HCT-116 cells (IC50: 5.42μg/mL). On the other hand, the n-hexane and EtOAc fractions demonstrated significant inhibitory activities against the Gram positive bacteria S. pyogenes and C. perfringens; and the Gram negative bacterium S. enteritidis. Our results warrant the therapeutic potential of S. irio as nutritional supplement to reduce the risk of contemporary diseases. Additionally, a validated high performance thin-layer chromatography (HPTLC) method was developed for the quantitative analysis of biomarker β-sitosterol glucoside (isolated in high quantity) from the n-hexane fraction. The system was found to furnish a compact, sharp, symmetrical and high resolution band for β-sitosterol glucoside (Rf=0.43±0.002). The limit of detection (LOD) and limit of quantification (LOQ) for β-sitosterol glucoside was found to be 21.84 and 66.18ngband−1, respectively. β-sitosterol glucoside was found to be present only in n-hexane fraction (2.10μg/mg of dried fraction) while it was absent in the other fractions of S. irio which validated the high cytotoxic and antibacterial activity of n-hexane fraction of S. irio

Portulaca oleracea Linn seed extract ameliorates hydrogen peroxide-induced cell death in human liver cells by inhibiting reactive oxygen species generation and oxidative stress

Purpose: To investigate the protective effects of Portulaca oleracea seed extract (POA) against cytotoxicity, oxidative stress and reactive oxygen species (ROS) generation induced by hydrogen peroxide (H2O2) in human liver cells (HepG2).Methods: The extract (POA) was obtained by ethanol extraction of P. oleracea seeds. Cytotoxicity in HepG2 cells was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, neutral red uptake (NRU) assay and morphological changes. The cells were pre-exposed to noncytotoxic concentrations (5 - 25 μg/mL) of POA for 24 h, and then cytotoxic (0.25 mM) concentration of H2O2. After 24 h of exposure, MTT and NRU assays were used to evaluate cell viability, while morphological changes were assessed using phase contrast inverted microscopy. The effect of POA on reduced glutathione (GSH) level, lipid peroxidation (LPO), and ROS generation induced by H2O2 was also studied.Results: The results showed that pre-exposure to POA (25 μg/mL) significantly (p <0.01) attenuated the loss of cell viability by up to 38 % against H2O2-induced oxidative stress and ROS generation. In addition, POA (25 μg/mL) significantly (p <0.01) increased GSH level (31 %), but decreased the levels of LPO (37 %) and ROS generation (49 %).Conclusion: This study demonstrates that POA has the capacity to protect HepG2 cells against H2O2- induced cell death by inhibiting oxidative stress and ROS generation.Keywords: Portulaca oleracea, HepG2 cells, Cytotoxicity, Oxidative stress, Reactive oxygen specie

Neoclerodane Diterpenoids from Reehal Fatima, <i>Teucrium yemense</i>

<i>Teucrium yemense</i> (Defl), locally known as Reehal Fatima, is a medicinal plant commonly grown in Saudi Arabia and Yemen. Phytochemical investigation of the aerial parts of <i>T. yemense</i> yielded six new neoclerodane diterpenoids, namely fatimanol A–E (<b>1</b>, <b>2</b>, <b>3</b>, <b>5</b>, and <b>6</b>) and fatimanone (<b>4</b>), and the known teulepicephin (<b>7</b>). As both the <i>Teucrium</i> genus and the related <i>Lamiaceae</i> family have previously been widely reported to possess anthelmintic and antimicrobial activities, the structural and biological characterization of the seven diterpenoids was pursued. The structures of the new compounds were elucidated from their 2D NMR and MS profiles and by comparison to related compounds. The structure of fatimanol D (<b>5</b>) was confirmed by X-ray crystallographic analysis. The new structures contribute to the breadth of knowledge of secondary metabolites in this genus

Chemical Composition, In Vitro Antimicrobial, Free-Radical-Scavenging and Antioxidant Activities of the Essential Oil of Leucas inflata Benth

The essential oil of Leucas inflata Balf.f. (Lamiaceae), collected in Yemen, was analyzed using gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) techniques. Forty-three components were recognized, representing 89.2% of the total oil. The L. inflata volatile oil was found to contain a high percentage of aliphatic acids (51.1%). Hexadecanoic acid (32.8%) and n-dodecanoic acid (7.8%) were identified as the major compounds. Oxygenated monoterpenes were distinguished as the second significant group of constituents (16.0%). Camphor (6.1%) and linalool (3.2%) were found to be the main components among the oxygenated monoterpenes. In addition, the volatile oil was assessed for its antimicrobial activity against four bacterial strains and one yeast species using broth micro-dilution assay for minimum inhibitory concentrations (MIC). In addition, antioxidant activity was measured utilizing the anti-radical activity of the sable free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-Carotene-linoleic acid assays. The oil of L. inflata showed an excellent antibacterial activity against only the tested Gram-positive bacteria with a MIC-value of 0.81 mg/mL. Furthermore, the oil demonstrated, at a concentration of 1 mg/mL, a weak to moderate antiradical and antioxidant activity of 38% and 32%, respectively

Artemisia monosperma induces ROS-mediated cell death in human colorectal carcinoma cells via modulating apoptotic genes

Background: Colorectal cancer (CRC) is the third most diagnosed cancer in the world. Artemisia monosperma (family: Asteraceae), a flowering plant species in the genus Artemisia, is widely used in traditional medicine. Plant extracts are well documented to combat against cancer diseases. Nevertheless, the outcome of A. monosperma extracts on human colorectal carcinoma cells (HCT-116) have not been investigated in details. Objectives: Thus, herein we aimed to evaluate the cytotoxic effect of different extracts i.e. n-hexane (AM-H), chloroform (AM-C), n-butanol (AM-B), and aqueous (AM-Aq) of A. monosperma against HCT-116 cells and to identify molecular mechanism(s) involved in cytotoxicity. Methods: The HCT-116 cells were treated with 10–1000 μg/mL of extracts for 24 h. Results: Our results displayed that all extracts significantly decreased cell viability in HCT-116 cells and changed the cellular morphology in a dose-dependent way. Among these extracts, AM-C showed higher cytotoxic activity with IC50 of 250.5 μg/mL. Further AM-C extract increased ROS production and reduced MMP level in HCT-116 cells. Moreover, AM-C extract induces cell apoptosis by the upregulation of proapoptotic related genes (p53, Bax, caspase-3, and caspase-9) and downregulation of antiapoptotic gene, Bcl-2. More significantly, these results revealed that AM-C extract enhances apoptosis in HCT-116 cells associated with ROS generation and mitochondrial dysfunction. Conclusion: Our findings suggest that the extracts of A. monosperma can be an auspicious therapeutic agent against colorectal cancer

Oxidative Stress Mediated Cytotoxicity, Cell Cycle Arrest, and Apoptosis Induced by Rosa damascena in Human Cervical Cancer HeLa Cells

Rosa damascena Mill (Damask rose), belonging to the Rosaceae family, is known for medicinal purposes in traditional medicine system. However, its anticancer activity has not been studied yet in detail. Herein, we aimed to investigate the cytotoxic effects of R. damascena hexane (RA-HE) and methanolic (RA-ME) extracts against human breast (MCF-7), lung epithelial (A-549), and cervical (HeLa) cancer cells. The RA-HE and RA-ME showed more potent cytotoxic effects against HeLa cells with an IC50 of 819.6 and 198.4 μg/ml, respectively. Further, cytotoxic concentrations of most effective extract (RA-ME) were used to evaluate the mechanism of cytotoxicity involved in HeLa cells. A concentration-dependent induction of lipid peroxidation (LPO) and reduction of glutathione (GSH) in HeLa cells treated with 250-1000 μg/ml of RA-ME confirms the association of oxidative stress. We also detected a noteworthy increase in reactive oxygen species (ROS) production and a decline in mitochondrial membrane potential (MMP) level in RA-ME-exposed HeLa cells. Flow cytometric data showed a strong dose-response relationship in cell cycle analysis between subG1 phase in HeLa cells and RA-ME treatment. Similarly, a concentration-dependent increase was recorded with Annexin V assay in HeLa cells going to late apoptosis. In conclusion, our findings suggest that RA-ME-induced cytotoxicity and apoptosis in HeLa cells are mediated by oxidative stress

Anticancer Potential of Green Synthesized Silver Nanoparticles Using Extract of Nepeta deflersiana against Human Cervical Cancer Cells (HeLA)

In this study, silver nanoparticles (AgNPs) were synthesized using aqueous extract of Nepeta deflersiana plant. The prepared AgNPs (ND-AgNPs) were examined by ultraviolet-visible spectroscopy, Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscope (SEM), and energy dispersive spectroscopy (EDX). The results obtained from various characterizations revealed that average size of synthesized AgNPs was 33 nm and in face-centered-cubic structure. The anticancer potential of ND-AgNPs was investigated against human cervical cancer cells (HeLa). The cytotoxic response was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), neutral red uptake (NRU) assays, and morphological changes. Further, the influence of cytotoxic concentrations of ND-AgNPs on oxidative stress markers, reactive oxygen species (ROS) generation, mitochondrial membrane potential (MMP), cell cycle arrest and apoptosis/necrosis was studied. The cytotoxic response observed was in a concentration-dependent manner. Furthermore, the results also showed a significant increase in ROS and lipid peroxidation (LPO), along with a decrease in MMP and glutathione (GSH) levels. The cell cycle analysis and apoptosis/necrosis assay data exhibited ND-AgNPs-induced SubG1 arrest and apoptotic/necrotic cell death. The biosynthesized AgNPs-induced cell death in HeLA cells suggested the anticancer potential of ND-AgNPs. Therefore, they may be used to treat the cervical cancer cells