An evaluation of the effects on the ovaries of hyperbaric oxygen therapy in a rat model of premature ovarian failure created with cyclophosphamide.

Objective: To evaluate hyperbaric oxygen therapy (HBO) based on ovarian histology, total antioxidant status (TAS), total oxidant status (TOS), and anti-müllerian hormone (AMH), in the ovarian insufiency (POI) model created with cyclophosphamide (CYP). Materials and Methods: The rats were separated into 3 groups of the control group (n=6), the CYP group (n=6), and the CYP+HBO group (n=6). The rats in the CYP group and the CYP+HBO group were injected intraperitoneally with 200 mg/kg CYP on day 1, followed by 8 mg/kg/day for 14 days to create POI. From the 15th day onwards, the rats in the CYP+HBO group were placed in a hyperbaric cabin and exposed to 100% oxygen at 2.4 atm pressure for one h, and were then returned to their cages at the end of the hour. Results: A statistically significant decrease was determined in the primordial and primary follicle counts in the CYP group compared with the control group (p<0.05). In the CYP+HBO group, a statistically significant increase was determined in the primordial and primary follicle counts (p<0.05). The serum AMH levels were seen to be significantly decreased in the CYP group compared with both the control group and the CYP+HBO groups. The HBO was seen to decrease TOS and increase TAS. Conclusion: HBO could be an alternative treatment to minimize the effect of ovarian follicle loss caused by CYP, which is used for treating tumors that commonly occur in young females of reproductive age.

Anti-inflammatory Effects of Pelargonium endlicherianum Fenzl. Extracts in Lipopolysaccharide-stimulated Macrophages.

OBJECTIVES: This study was designed to investigate the anti-inflammatory effects of Pelargonium endlicherianum Fenzl. and Pelargonium quercetorum Agnew. root extracts compared with the effects of commercial Pelargonium sidoides root extract by production of pro-inflammatory substances and inflammatory signal transduction on LPS-stimulated macrophages. MATERIALS AND METHODS: To measure the effects of root extracts on pro-inflammatory mediators, we used the following methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (cell viability or cytotoxcicity), enzyme-linked immunosorbent assay (cytokine production, prostoglandin E2 production), reverse transcriptase-polymerase chain reaction (COX-2, iNOS mRNA), Western blotting analysis [MAPK activation and NF-κB (p65) traslocation] and the Griess reaction (NO production). RESULTS: Stimulation of the RAW 264.7 cells with LPS (0.5 µg/mL, 6 hrs treatment) caused an elevated production of pro-inflammatory cytokines (TNF-α and IL-6), increased mRNA expression of COX-2 and inducible NO synthase with release of PGE2 and NO, activated MAPK (phosphorylation of c-Jun N-terminal kinase, extracellular signal-regulated kinase, P38) signalling pathway, and nuclear translocation of NF-κB (p65), which were markedly inhibited by the pre-treatment with 11% ethanol and 70% methanol root extracts of P. endlicherianum without causing any cytotoxic effects. P. quercetorum root extract only decreased TNF-α production and P. sidoides root extract alleviated P38/MAPK activation and COX-2 mRNA expression with PGE2 production. CONCLUSION: Our data indicate that especially 11% ethanol root extract of P. endlicherianum targets the inflammatory response of macrophages via inhibition of COX-2, IL-6, and TNF-α through inactivation of the NF-κB signalling pathway, supporting the pharmacologic basis of P. endlicherianum as a traditional herbal medicine for the treatment of inflammation and its associated disorders.

Aldose reductase inhibitory activity and antioxidant capacity of pomegranate extracts.

The pomegranate, Punica granatum L., has been the subject of current interest as a medicinal agent with wide-ranging therapeutic indications. In the present study, pomegranate ethanolic seed and hull extracts were tested, in comparison with a commercial sample, for the inhibition of aldose reductase, an enzyme involved in the etiology of diabetic complications. In vitro inhibition of rat lens aldose reductase was determined by a conventional method. Pomegranate ethanolic hull extract and commercial pomegranate hull extract exhibited similar aldose reductase inhibitory activity characterized by IC(50) values ranging from 3 to 33.3 µg/ml. They were more effective than pomegranate ethanolic seed extract with IC(50) ranging from 33.3 to 333 µg/ml. Antioxidant action of the novel compounds was documented in a DPPH test and in a liposomal membrane model, oxidatively stressed by peroxyl radicals. All the plant extracts showed considerable antioxidant potential in the DPPH assay. Pomegranate ethanolic hull extract and commercial pomegranate hull extract executed similar protective effects on peroxidatively damaged liposomal membranes characterized by 10

Investigation of ocular neovascularization-related genes and oxidative stress in diabetic rat eye tissues after resveratrol treatment.

Changes in vascular endothelial growth factor (VEGF), angiotensin-converting enzyme (ACE), matrix metalloproteinase (MMP)-9, and endothelial nitric oxide synthase (eNOS) mRNA expression profiles and oxidative stress in the eye tissue microenviroment may have important roles in ocular neovascularization and permeability in proliferative diabetic retinopathy. The present study investigated the effects of resveratrol (RSV) treatment on the mRNA expression profile of VEGF, ACE, MMP-9, and eNOS, which are associated with vascular neovascularization, and glutathione, protein carbonyl, and nitrite-nitrate levels, which are markers of oxidative stress in eyes of diabetic rats. Twenty-four Wistar albino male rats were divided into four groups. After diabetes induction with streptozotocin (10 mg/kg/day) RSV was administered to the RSV and diabetes mellitus (DM) + RSV groups for 4 weeks. The mRNA levels were measured by quantitative real-time polymerase chain reaction assay, and biochemical estimations were determined with spectrophotometric assays in eye homogenates. The mRNA expression levels of VEGF, ACE, and MMP-9 were increased in the DM group compared with the control group, and RSV treatment decreased their mRNA levels. Expression of eNOS mRNA was increased in the RSV and DM groups and decreased in the DM + RSV group. Nitrite-nitrate levels and protein carbonyl content were increased and glutathione levels were decreased in the DM group compared with controls. Consequently, these data suggest that RSV suppressed the expression of eNOS, which is actively involved in the inflammation and healing process in chronic diabetes. Although oxidative stress was increased in eye tissue from diabetic rats, mRNA levels of VEGF, MMP-9, and ACE genes associated with vascular remodeling did not change in diabetic eyes.

Polyphenolic extracts from Olea europea L. protect against cytokine-induced ß-cell damage through maintenance of redox homeostasis.

Various pancreatic ß-cell stressors, including cytokines, are known to induce oxidative stress, resulting in apoptotic/necrotic cell death and inhibition of insulin secretion. Traditionally, olive leaves or fruits are used for treating diabetes, but the cellular mechanism(s) of their effects are not known. We examined the effects of Olea europea L. (olive) leaf and fruit extracts and their component oleuropein on cytokine-induced ß-cell toxicity. INS-1, an insulin-producing ß-cell line, was preincubated with or without increasing concentrations of olive leaf or fruit extract or oleuropein for 24 hr followed by exposure to a cytokine cocktail containing 0.15 ng/mL interleukin-1ß (IL-1ß), 1 ng/mL interferon-γ (IFN-γ), and 1 ng/mL tumor necrosis factor-α (TNF-α) for 6 hr. The cytotoxicity was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) testing. Apoptosis was quantified by detecting acridine orange/ethidium bromide-stained condensed nuclei under a fluorescent microscope. The cells exposed to cytokines had a higher apoptotic rate, a decreased viability (MTT), and an increased caspase 3/7 activity. Both extracts and oleuropein partially increased the proportion of living cells and improved the viability of cells after cytokines. The protective effects of extracts on live cell viability were mediated through the suppression of caspase 3/7 activity. Oleuropein did not decrease the amount of both apoptotic and necrotic cells, whereas extracts significantly protected cells against cytokine-induced death. Cytokines led to an increase in reactive oxygen species (ROS) generation and inhibited glutathione level, superoxide dismutase activity, and insulin secretion in INS-1. Insulin secretion was almost completely protected by leaf extract, but was partially affected by fruit extract or oleuropein. Neither cytokines nor olive derivatives had a significant effect on cellular cytochrome c release and catalase activity. Moreover, the cells incubated with each extract or oleuropein showed a significant reduction in cytokine-induced ROS production and ameliorated abnormal antioxidant defense. The molecular mechanism by which olive polyphenols inhibit cytokine-mediated ß-cell toxicity appears to be involving the maintenance of redox homeostasis.

Novel hexahydropyridoindole derivative as prospective agent against oxidative damage in pancreatic ß cells.

The potential protective effect of (±)-8-methoxy-1,3,4,4a,5,9b-hexahydro-pyrido[4,3-b]indole-2-carboxylic acid ethyl ester (II) was assessed against hydrogen peroxide (H2O2)-cytotoxicity in rat pancreatic INS-1E ß cells and compared with the effect of the related pyridoindole, stobadine (I), a promising indole-type reference antioxidant. Only pre-treatment with the compound (II) led to a significant preservation of the metabolic and secretory functions of the cells exposed to H2O2. The caspase-9 and -3 activities, as well as the early apoptotic changes of plasma membrane, were suppressed in the cells pre-incubated with both of compounds tested. However, only pyridoindole (II) inhibited profoundly the time-delayed apoptotic changes, These results suggest that pyridoindole (II) characterized by enhanced intrinsic antioxidant efficiency, may protect ß cells against cytotoxic effects of H2O2, involved in the development of both type 1 and type 2 diabetes.