Interleukin 15 blockade protects the brain from cerebral ischemia-reperfusion injury.

Acute ischemic stroke is followed by a complex interplay between the brain and the immune system in which ischemia-reperfusion leads to a detrimental inflammatory response that causes brain injury. In the brain, IL-15 is expressed by astrocytes, neurons and microglia. Previous study showed that ischemia-reperfusion induces expression of IL-15 by astrocytes. Transgenic over-expression of IL-15 in astrocytes aggravates ischemia-reperfusion brain damage by increasing the levels and promoting the effector functions of CD8+ T and NK cells. Treatment of neonatal rats with IL-15 neutralizing antibody before hypoxia-ischemia induction reduces the infarct volume. However, as stroke-induced inflammatory responses differ between neonate and adult brain, the effects of IL-15 blockade on the injury and immune response arising from stroke in adult animals has remained unclear. In this study, we examined the effect of post-ischemia/reperfusion IL-15 blockade on the pathophysiology of cerebral ischemia-reperfusion in adult mice. Using a cerebral ischemia-reperfusion model, we compared infarct size and the infiltrating immune cells in the brain of wild type (WT) mice and Il15-/- mice lacking NK and memory CD8+ T cells. We also evaluated the effects of IL-15 neutralizing antibody treatment on brain infarct volume, motor function, and the status of brain-infiltrating immune cells in WT mice. Il15-/- mice show a smaller infarct volume and lower numbers of activated brain-infiltrating NK, CD8+ T, and CD4+ T cells compared to WT mice after cerebral ischemia-reperfusion. Post-ischemia/reperfusion IL-15 blockade reduces infarct size and improves motor and locomotor activity. Furthermore, IL-15 blockade reduces the effector function of NK, CD8+ T, and CD4+ T cells in the ischemia-reperfusion brain of WT mice. Ablation of IL-15 responses after cerebral ischemia-reperfusion ameliorates brain injury in adult mice. Therefore, targeting IL-15 is a potential effective therapy for ischemic stroke.

Protective effect of methyl gallate from Toona sinensis (Meliaceae) against hydrogen peroxide-induced oxidative stress and DNA damage in MDCK cells.

Methyl gallate (MG) has been shown to be an effective antioxidant in a variety of acellular experiments. Accordingly, this study was designed to assess the ability of MG, extracting from Toona sinensis to protect cultured Madin-Darby canine kidney (MDCK) cells against hydrogen peroxide (H2O2)-mediated oxidative stress. Trolox, a cell permeable and water-soluble vitamin E analogue, was included for comparison. First, when MDCK cells were pretreated with MG and trolox for 1 h, followed by exposing to H2O2 (0.8 mM) for an additional hour, we found that the intracellular peroxide productions, as reflected by dichlorofluorescein (DCF) fluorescence, were shown to be decreased in a concentration-dependent manner. Furthermore, using C11-BODIPY581/591 as a lipid peroxidation probe, we also found that MG, in a concentration of 100 microM, could alleviate lipid peroxidation of the cells exposed to a short-term H2O2 treatment. In addition, MG-treated cells could prevent intracellular glutathione (GSH) from being depleted following an exposure of H2O2 (8.0 mM) for a 3 h period. Next, we also examined the effect of MG on H2O2-mediated oxidative damage to DNA. Using 8-oxoguanine as an indicator for oxidative DNA damage, we demonstrated that the percentage of MDCK cells containing 8-oxoguanine was drastically increased by exposing to H2O2 (40 mM) for 3 h. However, 8-oxoguanine contents were shown to be significantly decreased in the presence of MG prior to H2O2 exposure. Comparatively, MG was shown to be a better protective agent against oxidative damage to DNA as compared to trolox. Taken together, our data suggest that MG is effective in preventing H2O2-induced oxidative stress and DNA damage in MDCK cells. The underlying mechanisms involved scavenging of intracellular reactive oxygen species (ROS), inhibition of lipid peroxidation and prevention of intracellular GSH depletion.

Folate deficiency triggers an oxidative-nitrosative stress-mediated apoptotic cell death and impedes insulin biosynthesis in RINm5F pancreatic islet ß-cells: relevant to the pathogenesis of diabetes.

It has been postulated that folic acid (folate) deficiency (FD) may be a risk factor for the pathogenesis of a variety of oxidative stress-triggered chronic degenerative diseases including diabetes, however, the direct evidence to lend support to this hypothesis is scanty. For this reason, we set out to study if FD can trigger the apoptotic events in an insulin-producing pancreatic RINm5F islet ß cells. When these cells were cultivated under FD condition, a time-dependent growth impediment was observed and the demise of these cells was demonstrated to be apoptotic in nature proceeding through a mitochondria-dependent pathway. In addition to evoke oxidative stress, FD condition could also trigger nitrosative stress through a NF-κB-dependent iNOS-mediated overproduction of nitric oxide (NO). The latter compound could then trigger depletion of endoplasmic reticulum (ER) calcium (Ca(2+)) store leading to cytosolic Ca(2+) overload and caused ER stress as evidence by the activation of CHOP expression. Furthermore, FD-induced apoptosis of RINm5F cells was found to be correlated with a time-dependent depletion of intracellular glutathione (GSH) and a severe down-regulation of Bcl-2 expression. Along the same vein, we also demonstrated that FD could severely impede RINm5F cells to synthesize insulin and their abilities to secret insulin in response to glucose stimulation were appreciably hampered. Even more importantly, we found that folate replenishment could not restore the ability of RINm5F cells to resynthesize insulin. Taken together, our data provide strong evidence to support the hypothesis that FD is a legitimate risk factor for the pathogenesis of diabetes.