Functionalized Fullerene Increases NF-κB Activity and Blocks Genotoxic Effect of Oxidative Stress in Serum-Starving Human Embryo Lung Diploid Fibroblasts

Abstract

The influence of a water-soluble [60] fullerene derivative containing five residues of 3-phenylpropionic acid and a chlorine addend appended to the carbon cage (F-828) on serum-starving human embryo lung diploid fibroblasts (HELFs) was studied. Serum deprivation evokes oxidative stress in HELFs. Cultivation of serum-starving HELFs in the presence of 0.1–1 µM F-828 significantly decreases the level of free radicals, inhibits autophagy, and represses expression of NOX4 and NRF2 proteins. The activity of NF-κB substantially grows up in contrast to the suppressed NRF2 activity. In the presence of 0.2–0.25 µM F-828, the DSB rate and apoptosis level dramatically decrease. The maximum increase of proliferative activity of the HELFs and maximum activity of NF-κB are observed at these concentration values. Conclusion. Under the conditions of oxidative stress evoked by serum deprivation the water-soluble fullerene derivative F-828 used in concentrations of 0.1 to 1 µM strongly stimulates the NF-κB activity and represses the NRF2 activity in HELFs.

Autophagy Flux Contributes to Regulation of Components of Eclipta prostrata L. on Cigarette Smoking-Induced Injury of Bronchial Epithelial Cells

“…The role of autophagy in the mechanisms of respiratory disease has been controversial with studies that have suggested both protective and injurious aspects ( Kim et al, 2016 ). Whether autophagy is protective or detrimental in respiratory disease seems to depend on the extent of its activation, specific stimuli/circumstances, and specific cell type ( Ershova et al, 2016 ). Additionally, the autophagic damage of NHBE cells was related to the stimulation time and concentration of CSE.…”

Effects of Functionalized Fullerenes on ROS Homeostasis Determine Their Cytoprotective or Cytotoxic Properties

“…Approval#5 was obtained from the Committee for Medical and Health Research Ethics of RCMG. Cells were seeded at 1.7 × 10 4 per mL in DMEM (Paneco, Moscow, Russia) with a 10% fetal calf serum (PAA Laboratories, Vienna, Austria), 50 U/mL penicillin, 50-μg/mL streptomycin and 10-μg/mL gentamycin (all the reagents were from Sigma-Aldrich, St. Louis, MO, USA) and cultured at 37 °C for 2 or 24 h as described elsewhere [ 35 , 36 ]. Fullerenes were dissolved under sterile conditions in a minimum volume of sterilized water or a saline solution.…”

“…Cells were grown in a 96-well plate for 72 h. Cell viability was assessed with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, as described previously [ 35 , 36 ]. The plates were read at 550 nm with EnSpire plate reader (EnSpire Equipment, Turku, Finland).…”

“…Cells were grown in slide flasks (25 cm 3 , Thermo Fisher Scientific, Waltham, MA, USA), fixed in 3% paraformaldehyde at 4 °C for 20 min, washed with PBS and then permeabilized with 0.1% Triton X-100 in PBS for 15 min at room temperature, followed by blocking with 0.5% BSA in PBS for 1 h and incubation overnight at 4 °C with the antibodies. After washing with 0.1% Triton X-100 in PBS, fibroblasts were incubated for 2 h at room temperature with the FITC goat anti-mouse IgG, washed with PBS and then stained with DAPI (4′,6-diamidino-2-phenylindole) (Sigma-Aldrich, Saint Louis, MO, USA) as described in [ 35 , 36 ].…”

Antioxidant Properties of Fullerene Derivatives Depend on Their Chemical Structure: A Study of Two Fullerene Derivatives on HELFs

“…Approval#5 was obtained from the Committee for Medical and Health Research Ethics of RCMG. Cells were seeded at 1.7 × 104 per ml in DMEM (Paneco, Moscow, Russia) with 10% fetal calf serum (PAA, Vienna, Austria), 50 U/ml penicillin, 50 μ g/ml streptomycin, and 10 μ g/ml gentamycin, and cultured at 37°C for 2 or 24 h as previously described in [13, 14]. Investigated fullerene derivatives were added to the medium, and the cells were cultured for periods ranging from 15 min to 48 h.…”