Oxidative Stress Evaluation in Ischemia Reperfusion Models: Characteristics, Limits and Perspectives.

Ischemia reperfusion injury is a complex process consisting of a seemingly chaotic but actually organized and compartmentalized shutdown of cell function, of which oxidative stress is a key component. Studying oxidative stress, which results in an imbalance between reactive oxygen species (ROS) production and antioxidant defense activity, is a multi-faceted issue, particularly considering the double function of ROS, assuming roles as physiological intracellular signals and as mediators of cellular component damage. Herein, we propose a comprehensive overview of the tools available to explore oxidative stress, particularly in the study of ischemia reperfusion. Applying chemistry as well as biology, we present the different models currently developed to study oxidative stress, spanning the vitro and the silico, discussing the advantages and the drawbacks of each set-up, including the issues relating to the use of in vitro hypoxia as a surrogate for ischemia. Having identified the limitations of historical models, we shall study new paradigms, including the use of stem cell-derived organoids, as a bridge between the in vitro and the in vivo comprising 3D intercellular interactions in vivo and versatile pathway investigations in vitro. We shall conclude this review by distancing ourselves from "wet" biology and reviewing the in silico, computer-based, mathematical modeling, and numerical simulation options: (a) molecular modeling with quantum chemistry and molecular dynamic algorithms, which facilitates the study of molecule-to-molecule interactions, and the integration of a compound in a dynamic environment (the plasma membrane...); (b) integrative systemic models, which can include many facets of complex mechanisms such as oxidative stress or ischemia reperfusion and help to formulate integrated predictions and to enhance understanding of dynamic interaction between pathways.

Protective effects of Iranian Achillea wilhelmsii essential oil on acetaminophen-induced oxidative stress in rat liver.

CONTEXT: Achillea wilhelmsii C. Koch (Asteraceae) is widely used in Iranian traditional medicine. OBJECTIVE: This in vivo study evaluates the hepatoprotective role of Iranian A. wilhelmsii oils against acetaminophen-induced oxidative damages in rats. MATERIALS AND METHODS: The animals were divided into five groups: in negative control and control groups, the DMSO and 500 mg/kg acetaminophen were i.p. injected, respectively. In treatment groups, 100 and 200 mg/kg oils and 10 mg/kg BHT were given i.p. immediately after acetaminophen administration. Then, the hepatic oxidative/antioxidant parameters such as lipid peroxidation (LP), glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), and ferric reducing ability of plasma (FRAP) were measured in time intervals (2, 4, 8, 16, and 24 h) after administrations confirmed by histophatological consideration at 24 h. RESULTS: The results indicated that acetaminophen caused a significant elevation in SOD activity (8-24 h) and LP and FRAP levels (4 h) paralleled with significant decline in GSH level (4 and 8 h). The apparent oxidative injury was associated with evident hepatic necrosis confirmed in histological examination. The presences of A. wilhelmsii oils (100 and 200 mg/kg) with acetaminophen mitigated significantly the rise in SOD, LP, and FRAP levels and restored the GSH compared with the group treated with acetaminophen. These were confirmed by histological examination indicating the hepatic necrosis reversal by the oils. DISCUSSION AND CONCLUSION: It can be concluded that concomitant administration of A. wilhelmsii oils with acetaminophen may be useful in reversing the drug hepatotoxicity.

Exploring the Impact of French Raw-Milk Cheeses on Oxidative Process Using Caenorhabditis elegans and Human Leukocyte Models.

Fermented foods, including cheeses, have garnered increased interest in recent years for their potential health benefits. This study explores the biological properties of eight French raw-milk cheeses-goat cheese, Saint-Nectaire, Cantal, Bleu d'Auvergne, Roquefort, Comté, Brie de Meaux, and Epoisses-on oxidative processes using both in vivo (Caenorhabditis elegans) and in vitro (human leukocytes) models. A cheese fractionation protocol was adapted to study four fractions for each cheese: a freeze-dried fraction (FDC) corresponding to whole cheese, an apolar (ApE), and two polar extracts (W40 and W70). We showed that all cheese fractions significantly improved Caenorhabditis elegans (C. elegans) survival rates when exposed to oxidative conditions by up to five times compared to the control, regardless of the fractionation protocol and the cheese type. They were also all able to reduce the in vivo accumulation of reactive oxygen species (ROS) by up to 70% under oxidative conditions, thereby safeguarding C. elegans from oxidative damage. These beneficial effects were explained by a reduction in ROS production up to 50% in vitro in human leukocytes and overexpression of antioxidant factor-encoding genes (daf-16, skn-1, ctl-2, and sod-3) in C. elegans.

Effects of FGF2 Priming and Nrf2 Activation on the Antioxidant Activity of Several Human Dental Pulp Cell Clones Derived From Distinct Donors, and Therapeutic Effects of Transplantation on Rodents With Spinal Cord Injury.

In recent years, the interest in cell transplantation therapy using human dental pulp cells (DPCs) has been increasing. However, significant differences exist in the individual cellular characteristics of human DPC clones and in their therapeutic efficacy in rodent models of spinal cord injury (SCI); moreover, the cellular properties associated with their therapeutic efficacy for SCI remain unclear. Here, using DPC clones from seven different donors, we found that most of the clones were highly resistant to H2O2 cytotoxicity if, after transplantation, they significantly improved the locomotor function of rats with complete SCI. Therefore, we examined the effects of the basic fibroblast growth factor 2 (FGF2) and bardoxolone methyl (RTA402), which is a nuclear factor erythroid 2-related factor 2 (Nrf2) chemical activator, on the total antioxidant capacity (TAC) and the resistance to H2O2 cytotoxicity. FGF2 treatment enhanced the resistance of a subset of clones to H2O2 cytotoxicity. Regardless of FGF2 priming, RTA402 markedly enhanced the resistance of many DPC clones to H2O2 cytotoxicity, concomitant with the upregulation of heme oxygenase-1 (HO-1) and NAD(P)H-quinone dehydrogenase 1 (NQO1). With the exception of a subset of clones, the TAC was not increased by either FGF2 priming or RTA402 treatment alone, whereas it was significantly upregulated by both treatments in each clone, or among all seven DPC clones together. Thus, the TAC and resistance to H2O2 cytotoxicity were, to some extent, independently regulated and were strongly enhanced by both FGF2 priming and RTA402 treatment. Moreover, even a DPC clone that originally exhibited no therapeutic effect on SCI improved the locomotor function of mice with SCI after transplantation under both treatment regimens. Thus, combined with FGF2, RTA402 may increase the number of transplanted DPCs that migrate into and secrete neurotrophic factors at the lesion epicenter, where reactive oxygen species are produced at a high level.

A combination containing natural extracts of clove, Sophora flower bud, and yam improves fertility in aged female mice via multiple mechanisms.

Introduction: With society development, the age at which women choose to have children has been gradually delayed. To improve the reduced fertility in women at advanced maternal age, we developed a combination containing natural extracts from clove, Sophora flower bud and Chinese yam with a mass ratio 15:6:10 and named it as DACHAO. Methods and Results: We then gavage DACHAO at a dose of 310 mg/kg BW to female mice at 10 month of age and investigated its effects on ovarian functions. Using MitoTracker probes, ROS, and JC-1 staining, we found that DACHAO treatment improved mitochondria functions in oocytes from aged mice. We also observed increased blastocyst formation when mature oocytes from control and DACHAO treated mice were for IVF and in vitro embryo culture. Cell counting and TUNEL assay further revealed increased cell numbers and decreased apoptosis in blastocysts of DACHAO group. After control or DACHAO treated mice being mated with fertile male mice, fertility test revealed a greater first litter size in the DACHAO group. Further studies demonstrated that DACHAO treatment could alleviate the retarded ovarian function in aged mice via changes in serum hormone levels, over-expression of antioxidant factors, under-expression of inflammation-related factors, and reduced apoptosis in the ovaries. Discussion: Thus, the new combination DACHAO will be a good choice in clinic to improve ovarian functions for women at advanced maternal age.

Atractylenolide II prevents radiation damage via MAPKp38/Nrf2 signaling pathway.

Ionizing radiation (IR) can act as a negative factor for human homeostasis, by causing and even aggravating a series of pathological conditions. To protect the intactness of normal tissues, effective anti-radiation drugs are urgently needed for alleviating the outcomes of radioactive damage. In this study, we demonstrate that atractylenolide II (ATR II), a sesquiterpenoid monomer extracted from traditional Chinese medicine atractylodes macrocephala, can markedly suppress IR damage by promoting the expression of antioxidant factors heme oxygenase-1 (HO-1) and NAD(P)H dehydrogenase quinone oxido-reductase 1 (NQO-1), which are mediated by nuclear factor-erythroid 2-like 2 (Nrf2) signaling pathway. Furthermore, here we reveal that ATR II effectively upregulates the expression of mitogen-activated protein kinase p38 (MAPKp38), which also acts as a regulator of Nrf2 signaling cascade. Indeed, treatment with a MAPKp38 inhibitor can significantly downregulate the expression of Nrf2 and its downstream target genes HO-1 and NQO-1 and, consequently, abolish the protective effect of ATR II against IR. Consistently, ATR II also has a protective function against IR-induced damage in animal models. In conclusion, our study provides an unexpected function of ATR II in preventing IR-induced damage by modulating MAPKp38/Nrf2 signaling pathway.

p-Nonylphenol Impairment of Osteogenic Differentiation of Mesenchymal Stem Cells was Found to be Due to Oxidative Stress and Down-Regulation of RUNX2 and BMP.

OBJECTIVES: Previously, it was found that the para-nonylphenol (p-NP) impairs the osteogenic differentiation of rat bone marrow mesenchymal stem cells (rBMSCs); thus the aim of the present study was to evaluate the mechanism of the impairment. METHODS: rBMSCs after 3rd passage cultured in osteogenic media in the presence of 0, 0.5 and 2.5 µM p-NP for 5, 10, 15 and 20 days. The study investigated the viability of the cells using MTT assays. The mineralization was studied using Alizarin red quantification analysis. Using a flame-photometer, the electrolytes (sodium and potassium) were measured, and the level of calcium as well as ALT, AST, ALP and LDH was determined by commercial kits. The level of total-antioxidant, MDA and the activity of SOD and CAT were estimated with the help of a spectrophotometer. Gene expression was studied using rt-PCR. RESULTS: The p-NP treatment of osteogenic differentiated MSCs showed intracellular electrolyte imbalance and variation of cellular metabolism. In addition, we observed oxidative stress due to the reduction of total antioxidant capacity and the imbalance of antioxidant enzymes activity. Investigating the genes involved in the osteogenic differentiation of MSCs to osteoblast showed that the 2.5 µM of p-NP reduced the expression of the ALP, SMAD, BMP and RUNX2 genes. CONCLUSION: The study concludes that this pollutant via influencing the genomics and metabolic imbalance, as well as oxidative induction, caused a reduction of mineralization and differentiation of MSCs. This environmental pollutant might cause osteoporosis, which necessitates raising public awareness, especially to those who live in the industrial area to prevent its drastic effect.

Comparison of honey and dextrose solution on post-operative peritoneal adhesion in rat model.

BACKGROUND: Peritoneal adhesion between abdominal organs is a complication of surgery. It causes major complications like pain, bowel obstruction, infertility and increases risk of death. Honey is known to have anti-inflammatory and antioxidant properties potentially relevant for adhesive protection. METHODS: Thirty rats were divided into five groups: negative control without any surgical procedure (normal group), control group treated with normal saline, experimental group I treated with 1ml of 10% honey, experimental group II treated with honey at half concentration of group I (honey0.5), and positive control group receiving 1ml of dextrose 5%. Inflammatory, growth and angiogenesis factors (TNF-α, Il-6, IL-1ß, TGF-ß1 and VEGF) of the adhesion tissue were assessed using ELISA. Antioxidant factors (NO, GSH and MDA) were also assessed using biochemical procedures. RESULTS: The difference between peritoneal adhesion scores, TNF-α, IL-1ß, IL-6, TGF-ß1, VEGF, NO, GSH and MDA value of all groups was strongly significant (p<0.001). We showed that honey can decrease peritoneal adhesion (p<0.001), TNF-α (p<0.001), IL-1ß (p<0.001), IL-6 (p<0.001), TGF-ß1 (p<0.001), VEGF (p<0.001), NO (p<0.001), MDA (p<0.001) and increase GSH (p<0.001) compared with control group. Honey 0.5 also significantly decreased peritoneal adhesion (p<0.001), TNF-α (p<0.001), IL-1ß (p<0.01), IL-6 (p<0.001), VEGF (p<0.001), NO (p<0.001), MDA (p<0.01) and increase GSH (p<0.001) compared with control group. CONCLUSIONS: We find that honey can decrease inflammatory, growth and angiogenesis factors which can advance peritoneal adhesion and increase antioxidant factors. Honey could serve as a protective agent for peritoneal adhesion.