The transient receptor potential vanilloid 4 (TRPV4) is a nonselective cation channel involved in physical sensing in various tissue types. The present study aimed to elucidate the function and expression of TRPV4 in colonic vascular endothelial cells during dextran sulphate sodium (DSS)-induced colitis. The role of TRPV4 in the progression of colonic inflammation was examined in the 2% DSS-induced murine colitis model using immunohistochemical analysis, Western blotting, and Evans blue dye extrusion assay. DSS-induced colitis was significantly attenuated in TRPV4-deficient (TRPV4 KO) mice when compared to wild-type mice. Repeated intrarectal administration of GSK1016790A, a TRPV4 agonist, exacerbated the severity of DSS-induced colitis. Bone marrow transfer experiments demonstrated a dominant role of TRPV4 in non-haematopoietic cells for DSS-induced colitis. DSS treatment upregulated TRPV4 expression in the vascular endothelia of colonic mucosa and submucosa. DSS treatment increased vascular permeability, which was abolished in TRPV4 KO mice. The DSS-induced increase in vascular permeability was further enhanced by intravenous administration of GSK1016790A, which was abrogated by a TRPV4 antagonist RN1734. TRPV4 was co-localized with vascular endothelial (VE)-cadherin, and VE-cadherin expression was decreased by repeated intravenous administration of GSK1016790A during colitis. Furthermore, TRPV4 activation by GSK106790A decreased VE-cadherin expression in mouse aortic endothelial cells exposed to TNF-α. These findings indicate that TRPV4 upregulation in vascular endothelial cells contributes to the progression of colonic inflammation via the activation of vascular permeability. Thus, TRPV4 is an attractive target for the treatment of inflammatory bowel diseases.

Eosinophilic esophagitis (EoE) and eosinophilic gastroenteritis (EGE), which are included in eosinophilic gastrointestinal disorders (EGIDs), are allergic gastrointestinal diseases mainly caused by food allergens, which features dense infiltration of eosinophiles in the gastrointestinal mucosa. A possible mechanism of these diseases are Th2 type allergic reactions, including IL-5, IL-13, and IL-15, thymic stromal protein (TSLP), and eotaxin 3, which are considered to have important roles. The number of patients with EoE is rapidly increasing in both Western and Asian countries. In Japan, a research group of the Ministry of Health, Labor and Welfare has announced recommendations for the diagnosis and management of EoE and EGE in 2015. For a diagnosis of EoE, endoscopic abnormalities and histological confirmation of dense eosinophile infiltration in the esophageal epithelial layer are important, in addition to identifying dysphagia symptoms. As for eosinophilic gastroenteritis, blood test findings are more useful and the role of an endoscopic examination is reduced. Glucocorticoid administration is standard treatment for these diseases, while proton pump inhibitors are frequently effective for EoE. As for EoE, on the one hand, a variety of studies have been performed and new findings collected. On the other hand, information concerning EGE is limited and additional evidence is needed to establish effective treatment options.

Reactive oxygen species (ROS) are known to play a critical role in the development of non-alcoholic steatohepatitis (NASH). To clarify the source of ROS, we examined the expression of superoxide-generating NADPH oxidase isoforms in the liver of high-fat and high-cholesterol (HFC) diet-fed mice. The mRNA expression of NOX1 was significantly elevated in mice on HFC diet for 8 weeks. Increased levels of serum alanine aminotransferase and hepatic cleaved caspase-3 in HFC diet-fed wild-type mice (WT) were significantly ameliorated in mice deficient in Nox1 (Nox1-KO). Increased nitrotyrosine adduct formation, a marker of peroxynitrite-induced injury, was observed in hepatic sinusoids of WT, which was significantly suppressed in NOX1-KO. NOX1 mRNA was mainly expressed in liver sinusoidal endothelial cells (LSECs), and it was significantly up-regulated in primary cultured LSECs treated with palmitic acid (PA). The production of nitric oxide by LSECs and LSECs-dependent relaxation of hepatic stellate cells were significantly attenuated by PA treatment. In contrast, these effects of PA were not observed in cells isolated from Nox1-KO. Taken together, the up-regulation of NOX1 in LSECs may elicit peroxynitrite-mediated cellular injury and impair hepatic microcirculation through reduced bioavailability of nitric oxide. ROS derived from NOX1 may therefore constitute a critical component in the development of NASH.

Nonalcoholic fatty liver disease (NAFLD), especially nonalcoholic steatohepatitis (NASH), is a disease that may lead to liver cirrhosis or hepatocellular carcinoma. The number of patients is increasing steadily and the pathology is diverse. Effective treatment is still mainly focused on diet and exercise therapy. With the advent of the next-generation sequencer, the intestinal flora in NAFLD/NASH is being elucidated. We believe that elevation of endotoxin in the blood caused by intestinal bacterial dysbiosis caused by intestinal permeability enhancement is important for NASH pathology and control of intestinal permeability enhancement has a new possibility for treatment of NAFLD/NASH.

Posttraumatic stress disorder (PTSD) is most often induced by traumatic events and serious public health problems. PTSD is characterized by excessive response to contextual memory and impaired fear extinction and also associated with mild cognitive impairment, attention and learning deficits. Clinical and animal studies suggest that increased susceptibility of emotion- and fear-related neuronal circuits, including those in the amygdala, prefrontal cortex and hippocampus, contributes to development and retention of PTSD symptoms. However, mechanisms underlying this susceptibility to fear are not known and the useful therapeutic approaches are limited. Recently, there have been reports that ω3 LCPUFA supplementation can prevent development of PTSD and significantly ameliorate symptoms in patients with PTSD after accidental injury such as motor vehicle accidents and natural calamities. Importantly, Fabp7 null mice exhibit enhancement of fear memory consolidation and anxiety-related behaviors that resemble PTSD-like behaviors in humans. In this review, we focused behavioral phenotype of PTSD in Fabp3 null mice. The Fabp3 null mice exhibit cognitive deficits, hyperlocomotion and impaired fear extinction, and thus show PTSD-like behaviors. Chronic administration of ramelteon, a melatonin receptor agonist, improved all PTSD-like behaviors tested in Fabp3^－/－ mice. Relevant to mechanisms underlying impaired fear extinction, we observed that Ca^2＋/calmodulin-dependent protein kinase II (CaMKII) autophosphorylation increases in the basolateral amygdala (BLA) but remained unchanges in the hippocampus of Fabp3^－/－ mice. Likewise, the number of c-Fos positive neurons in BLA significantly increased after exposure to contextual fear conditions. Finally, chronic ramelteon administration restored abnormal c-Fos expression and CaMKII autophosphorylation in the BLA of Fabp3^－/－ mice. Taken together, Fabp3^－/－ mice show PTSD-like behaviors, and ramelteon is an attractive candidate for PTSD therapeutics in human.

The use of minipigs has been increasing in the areas of pharmacology researches and drug development. The microminipig developed by Fuji Micra Inc. (Shizuoka, Japan) inherits characteristics of other pig strains showing several similarities to humans in anatomy, physiology, omnivorousness and diurnal, but at the same time has several advantages over other pig strains because of its small size which allows easy keeping, handling and dosing, and saving of test substances. The microminipig weighs about 10 kg at the age of 6 months. Canine cages can be used to keep the animal. Swine leukocyte antigens (SLA) are defined in each individual animal which is useful for testing immunological reactions. As there are many similarities in metabolic enzymes and transporters to those in humans, the microminipig is a powerful animal model for toxicokinetic studies. Unfortunately as in other minipigs the microminipig is not appropriate for embryo-fetal development studies of antibody drugs due to its poor placental transfer, but can be used for other reproductive and developmental studies. Repeat dose toxicity, safety pharmacology, immunotoxicity and local tolerance studies should be also other arenas of this animal model.