Hydrogen Peroxide-Induced Oxidative Damage in Human Chondrocytes: The Prophylactic Effects of Hypericum Perforatum Linn Extract on Deoxyribonucleic acid Damage, Apoptosis and Matrix Remodeling by a Disintegrin-Like and Metalloproteinase With Thrombospondin Motifs Proteinases

Abstract

Objectives: This in vitro study aimed to examine the protective roles of Hypericum perforatum Linn (HPL) extract on cell viability, DNA damage, apoptosis and a disintegrin-like and metalloproteinase with thrombospondin motifs (ADAMTS) proteins in chondrocytes induced by hydrogen peroxide (H2O2), as a model of chondrocytes subjected to reactive oxygen species (ROS) attack in rheumatoid arthritis and osteoarthritis. Materials and methods: Human chondrosarcoma cell line (OUMS-27) was used. Cells were incubated with different concentrations of methanolic extract (100, 400, and 750 μg/ml) of HPL for 36 hours, and then treated with 0.7 mM H2O2 for two hours. Trypan blue was used for evaluation of cell viability, while DNA damage was evaluated by alkaline Comet assay. Caspase-1, ADAMTS5, ADAMTS9, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) proteins were analyzed by Western blot. Results: In vitro H2O2 treatment decreased OUMS-27 cell viability. Cells pretreated with HPL at concentration of 400 μg/mL were best protected from H2O2 toxicity. Compared to 100 μg/ml concentration, pretreatment of cells with 750 or 400 μg/ml of HPL generated more protection against H2O2-induced DNA damage. Hydrogen peroxide application to the cells led to a slight increase in Caspase-1 expression, which shows no apoptosis. The most prominent increase in Caspase-1 level was shown in cells treated with 400 μg/ml of HPL extract. There was an increase in ADAMTS9 and a decrease in ADAMTS5 levels upon H2O2 administration. Pretreatment with HPL led to more decrease in ADAMTS5 level, indicating the protection of extracellular matrix attacked by these proteinases in cartilage tissue. Conclusion: It can be concluded that HPL has a potential to reverse the negative effects and processes induced by H2O2 in OUMS-27 cells and it can protect the surrounding cartilage area of chondrocytes from oxidative damage, which is suggested to be one of the main molecular factors accused for progression of rheumatoid arthritis and osteoarthritis.

Nettle (Urtica dioica L.) extracts as functional ingredients for production of chocolates with improved bioactive composition and sensory properties

“…SW480 cells were seeded in wells (2·10 4 per well) of 96 well black plates and subsequently treated with different concentrations of extract (0.5-5 x of the concentration present in originally prepared extract) for 1 and 2 h, with and without subsequent recovery period. Since H 2 O 2 is an important ROS which acts as a pro-oxidant in the cells affected by a certain imbalance, it initiates a cascade of reactions leading to oxidative stress related changes in the cells (Akyol et al 2014). In order to determine the nettle extract's antioxidative/prooxidative nature in the cells in which oxidative stress is already induced, cells were treated with 250 μM H 2 O 2 for 30 min, in order to induce oxidative stress.…”

Untitled

“…H 2 O 2 that induces oxidative damage(Akyol et al, 2014) led to a reduction in the viability of NRCM in this study. High levels of ROS production, as indicated by increased staining by carboxy-H 2 DCFDA and DHE, may lead to lipid peroxidation as well as cell membrane damage.…”

Regeneration and healing of bone and cartilage in type-1 and type-2 diabetes: the effects of insulin

“…Many studies are focused on the effects of several molecules to elucidate the matrix biology of chondrocytes and pathophysiology of experimental osteoarthritis (Akyol et al, 2013(Akyol et al, , 2014b. A list of the beneficial effects of insulin (Akyol et al, 2015a;Altuntaş et al, 2015) discussed in detail via whether its regulating effect on hyperglycemia or its direct effect on cells may be as follows: accelerating fracture healing, improving resistance to fractures, providing growth and development of skeletal structures in children, and ensuring success with preoperational processes in dental implant surgery.…”