The purpose of this study was to evaluate the efficacy of fish oil supplementation added to usual dry eye treatment in dry eye subjects in a randomized controlled trial. Twenty-seven typical dry eye subjects were selected from 43 candidates by the diagnostic criterion for dry eye in this study. They were assigned to the randomized fish oil group (n = 15) or the placebo group (n = 12). Fish oil group ingested fish oil capsules containing eicosapentaenoic acid (EPA, 1245 mg/day) and docosahexaenoic acid (DHA, 540 mg/day) for 12 weeks. Placebo group ingested placebo capsules without EPA or DHA. A visual analog scale test estimating subjective symptoms, the Schirmer I test, tear film break-up time (BUT) measurement, fluorescein staining, and rose bengal staining were performed every 4 weeks during the 12-week supplementation period and 4-week washout period. The subjective symptom of “eye pain”, BUT, and changes in rose bengal staining score of the fish oil group were significantly improved after 8-12 weeks of supplementation and/or 4 weeks of washout, compared to those of the placebo group. These results suggest that fish oil supplementation added to usual care may be effective in the treatment of dry eye.
2-deoxy-D-glucose (2DG) has been clinically evaluated for its potential use as an anticancer drug. Although 2DG is generally thought to inhibit the glycolytic pathway through accumulation of 2-deoxy-D-glucose-6-phosphate (2DG6P), it may also interfere with various other biological processes. Here, to further understand the role of 2DG as an inhibitor of tumor progression, we assessed the metabolism of 2DG in a human endometrial cancer cell line using capillary electrophoresis-time-of-flight mass spectrometry (CE-TOFMS). A total of 113 target metabolite peaks were identified and 90 metabolites of them were quantified. Furthermore, we present a new methodology which uses CE-TOFMS metabolome profiling following introduction of an artificial metabolite to evaluate tumor-specific metabolite traces. Aside from 2DG6P, we detected the presence of unique 2DG-derived deoxy metabolites in 2DG-treated cells. These metabolites may be responsible for the alteration of global metabolism in cells and act as various biological effectors.
The laminar formation in the developing cerebral cortex requires precisely regulated generation of phenotype-specific neurons. To determine whether neurotrophin-3 (NT3) is involved in this formation, we investigated the effects of NT3 administration in the telencephalic ventricular space on 13.5-day-old mouse embryos. NT3 increased the number of newly generated neurons and altered the neuronal phenotypes in the position and the transcription factors-expression profiles; the neuronal phenotypes originally committed for layer IV neurons were altered toward for layers II/III neurons. The former effects were observed when the parent progenitor cells were exposed to NT3 in the G1- to S-phase, whereas the latter effects were observed with exposure in the G1-phase. In addition, in vitro experiments revealed that the laminar fate alteration by NT3 was observed in the dissociated primary culture of cortical progenitors and the NT3 actions were suppressed by cotreatment with the MEK/ERK inhibitor. These observations suggest that NT3 is involved in the laminar formation of the developing cerebral cortex through the intercellular MEK/ERK pathway.
Chronic injury and inflammation in the liver are associated with the development of liver fibrosis. Expressions of transforming growth factor-β1 (TGF-β1) and hepatocyte growth factor (HGF) participate in the development and suppression, respectively, of liver fibrosis. Here, we investigated the effect of ONO-1301, a synthetic prostaglandin I2/IP receptor agonist, on liver fibrosis and on changes in the hepatic expressions of genes that regulate the progression of fibrosis in mice. Liver fibrosis was caused by the repetitive administration of CCl4 for 12 weeks, with ONO-1301 being administered during the last 4 weeks. The expressions of fibrogenic genes: TGF-β1, connective tissue growth factor, α-smooth muscle actin, type-I collagen, and type-III collagen were upregulated by chronic liver injury, which was associated with the expansion of myofibroblasts and the development of liver fibrosis. Treatment with ONO-1301 increased hepatic HGF mRNA expression, but decreased the expressions of TGF-β1, connective tissue growth factor, α-smooth muscle actin, and type-I and type-III collagen, which was associated with the suppression of myofibroblast expansion and liver fibrosis. Neutralizing antibody for HGF significantly attenuated the suppressive action of ONO-1301 on liver fibrosis and fibrogenic gene expressions. The therapeutic action of ONO-1301 on liver fibrosis may have occurred partly through HGF-mediated pathways.
Denervation alters the metabolism of the extracellular matrix (ECM) in skeletal muscle; however, the underlying mechanisms of ECM remodeling are not fully understood. The aim of this study was to elucidate the dynamic features of the ECM regulatory process in the early phase of denervated skeletal muscle in male Wistar rats. We investigated the expression of collagens (total, type I, and type III), transforming growth factor beta 1 (TGF-β1), and matrix metalloproteases (MMPs) together with their endogenous inhibitors (TIMPs), at the mRNA and/or protein level in the soleus muscles of control animals and at days 3, 7, and 14 post-denervation. Expression of mRNA encoding collagens was decreased at days 3 and 7, and had recovered by day 14, in parallel with total collagen protein content. Content of TGF-β1 protein was elevated sequentially, up to a maximum of 158% at day 14 post-denervation (P < 0.05), as was TIMP-2 mRNA expression (272% at day 14), whereas MMP-1, MMP-2, and TIMP-1 mRNA expression was not affected at any stage. The initial reduction of collagen mRNA may be responsible for hypoactivity caused by the disappearance of contractile function. Recovery of collagen mRNA/protein at day 14 may be due mainly to the suppressive effects of TGF-β1 on collagen degradation via TIMP-2 upregulation.
Brain-derived neurotrophic factor (BDNF) is considered to be involved in the etiology and treatment of stress-related mood disorders including anxiety. However, application of BDNF itself has many technical and pharmacological problems such as instability and/or a lack of appropriate delivery systems. To overcome these, we developed trans-2-decenoic acid ethyl ester (DAEE) as a stable and small molecule with BDNF-like activities. In the present study, we tested the activities of DAEE on a stress-induced anxiety-like mouse model. Mice were kept in cages and subjected to 3 sets of sequential leaning, drenching, and rotation as chronic mild stresses applied for 1-2 days over a 3-week period; and the anxiety-like symptom (reduced time spent in open arm of the maze) was evaluated by use of the elevated plus-maze test. A daily intraperitoneal administration of DAEE competed against the expression of the anxiety-like symptom when administered during the stress-loading period, and became therapeutic when administered after the stress-loading. This activity was accompanied by amelioration of the stress-induced reduction in the levels of BDNF and neurotrophin-3 mRNAs and phosphorylated extracellular signal-regulated kinases (ERK) 1/2 in the hippocampus. These results demonstrated that DAEE behaved like an anxiolytic and ameliorated this characteristic anxiety-like symptom, suggesting that DAEE may be a promising candidate for a novel anxiolytic with a new mechanism of action.
In the present study, we investigated the role of p16INK4a in the inhibition of DNA synthesis stimulated by hepatocyte growth factor (HGF) or epidermal growth factor (EGF) using RNA interference in primary cultured rat hepatocytes. The transfection of small interfering RNAs targeting p16INK4a reduced the corresponding mRNA and protein expression by more than approximately 90% and 50%, respectively, at 24 h after transfection. In the cells transfected with p16INK4a small interfering RNA, control, HGF, and EGF-stimulated DNA synthesis as assessed by 3H-thymidine incorporation increased by approximately 1.5-fold, 1.6-fold, and 1.7-fold, respectively, compared with that in the control small interfering RNA-transfected cells. These findings indicate that p16INK4a plays a significant role in the inhibition of DNA synthesis.