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

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

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

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

Erratum:Compounds Related to Saudin and Three New Series of Diterpenoids from Clutia lanceolata (Journal of Natural Products (2023) 86: 5 (1129−1149) DOI: 10.1021/acs.jnatprod.2c00761)

Re-examination and reinterpretation of the NOESY NMR data for 11 and 12 has led to reassignment of the relative stereochemical configurations at C-5 and C-10 for both compounds. The text discussing the relative configurations for compound 11 (lanceolide K) (p 1139) should be replaced with the following: A strong NOESY cross-peak between H-5 and H3-18 established these as being cis (β-oriented) on the cyclohexane ring. Thus, the γ-lactone ring was established as being fused cis on the cyclohexane ring. A NOESY cross-peak was also observed from H-5 to H-6Pro‑R (δH 4.23) (β-oriented) but not to H-6Pro‑S (δH 4.27) (α-oriented), helping to assign the signals for these diastereotopic protons. In the δ-lactone ring, a strong NOESY interaction between H-8 and H-12 indicated that these are both cis and that the ring must be in a (flattened) conformation for this strong prow−stern interaction. Thus, the methyl at C-8 and the furan ring at C-12 must both be on the β-face (the configuration at C-12 having been assumed from earlier analogues). Strong NOESY cross-peaks were also evident from both H-8 and H-12 to H-9, so H-9 was assigned as αoriented, and the bond from C-9 to the cyclohexane must be βoriented. This single C−C bond allows rotation, and so the other NOESY interactions between the two six-membered rings would be reasonable (e.g., H3-20 to H3-17, H-8, H-9, and both H2-11; H-5 to H-8). The NOESY cross-peak from H-5 to only one H-11 proton signal (δH 1.98) shows that this proton is on the β-face of the δ-lactone. The MM2 minimized model has this proton equatorial, which is consistent with its signal having only one large coupling (Jgem = 9.9 Hz). Thus, H-11 (δH 1.89) is αoriented/axial, which was supported by its signal having three large couplings (Jgem and 2 × Jax‑ax). The text discussing the relative configurations for compound 12 (lanceolide L) (pp 1140−1141) should be replaced with the following: The configuration of 12 was assigned using MM2 minimized models of the various possible diastereoisomers, the NOESY spectrum, and 1 H−1 H coupling constants. In the cyclohexane/γ-lactone bicyclic ring, H-19 (δH 3.79) gave strong NOESY cross-peaks with H3-18 and with H-5. Accordingly, H-5, H-19Pro‑S (δH 3.79), and Me-18 were all found to be cis to each other and on the β-face. This placed H-19Pro‑R (δH 4.09) on the α-face, and a strong NOESY cross-peak to H3-20 located the latter on the α-face also. In the δ-lactone, a NOESY connection was seen between H-8 and H-12, showing that they were cis (and α-oriented); so, the furan ring and Me-17 are β-oriented. The models used suggested H-11 (δH 1.96) and H-12 as being almost eclipsed in the deformed boat conformation of the δ-lactone ring. This was consistent with the former resonating as a broad quartet with a J value appropriate for being eclipsed with H-9 and H-12. Thus, H-11 (δH 2.12) was assigned as being on the βface. NOESY interactions were observed from H-9 to H-8 and to H-11 (δH 1.96), confirming that all are on the α-face. The absolute configuration of compound 12 was assigned by analogy with those of 1 and 2. Figure 1 should be replaced to show the correct configurations of compounds 11 and 12. We are grateful to Prof. Chao Li (National institute of Biological Sciences, Beijing, China) for bringing this to our attention initially. (Figure Presented).</p

Antioxidant and Anticancer Efficacies of Anethum graveolens against Human Breast Carcinoma Cells through Oxidative Stress and Caspase Dependency

Anethum graveolens, belonging to the family Apiaceae, has been extensively used for medicinal and therapeutic purposes since long. Plants encompass rich number of effective constituents with less toxicity. Thus, nowadays, the attempts are being made to search plant constituents that can prevent and reverse the chronic diseases, such as cancer. In this study, an in vitro antioxidant and anticancer efficacies of Anethum graveolens (AG-ME) were studied on human breast (MCF-7), lung (A-549), and cervical (HeLa) carcinoma cell lines. The antioxidant efficacies of AG-ME were evaluated by total antioxidant, DPPH radical scavenging, H2O2 scavenging, and ferrous reducing antioxidant assays. Further, the anticancer potential of AG-ME was also determined against different cancer cell lines. The AG-ME exhibited strong antioxidant activities as observed by antioxidant assays. AG-ME also showed a dose-dependent anticancer/cytotoxic potential against MCF-7, A-549, and HeLa cell lines. The AG-ME-induced reduction in GSH and increase in SOD activities indicates the role of oxidative stress in AG-ME-induced MCF-7 cell death. The results also exhibited that AG-ME triggered ROS production and significantly reduced MMP level. Moreover, a dose-dependent increase in caspase-3 and caspase-9 activities suggests that the AG-ME-induced MCF-7 cell death is caspase-dependent. Together, the present study provides reasoning and reassurance for the uses of A. graveleons for medical purposes as an antioxidant and anticancer agent. Additional investigations are required to examine biological and anticancer activities under an in vivo system to discover a possible beneficial use of AG-ME against diseases