Hisanori Kato

Molecular basis of the alteration in skin collagen metabolism in response to in vivo dexamethasone treatment: effects on the synthesis of collagen type I and III, collagenase, and tissue inhibitors of metalloproteinases

Summary Background Glucocorticoids are widely used for the treatment of various diseases, despite known side‐effects such as skin atrophy. Many studies have shown that the status of collagen fibres in the skin is affected by glucocorticoid treatment. However, the molecular mechanism underlying the alteration of collagen metabolism in the skin by glucocorticoid treatment remains unknown.

Effect of dietary proteins on insulin-like growth factor-1 (IGF-1) messenger ribonucleic acid content in rat liver

Effects of quantity and quality of dietary proteins on plasma immunoreactive insulin-like growth factor-1 (IGF-1) concentration, and content of IGF-1 mRNA in rat liver were investigated in rats. Plasma immunoreactive IGF-1 concentration in rats given a casein diet was higher than that in rats given a soya-bean-protein or protein-free diet. The IGF-1 mRNA content in liver was estimated by the Northern blot hybridization technique employing 32P-labelled rat IGF-1 complementary DNA (cDNA). At least four molecular species of IGF-1 mRNA of different molecular weight were found in rat liver. The sizes were 0.8-1.2, 2.0, 3.6-4.0 and 7.4 kb. Most of the mRNA species decreased in the livers of rats given a gluten diet (120 g gluten/kg diet) compared with rats given the casein diet. In particular, mRNA of 7.4 kb decreased markedly. When rats were fed on the protein-free diet, mRNA of all species decreased significantly. The estimated IGF-1 mRNA in the livers of rats fed on the gluten or protein-free diet was almost 0.4 of that of the rats given the casein diet. Feeding the soya-bean-protein diet did not result in a marked effect on the hepatic content of mRNA species of IGF-1. The results showed that liver IGF-1 mRNA content is sensitively regulated by quantity and nutritional quality of dietary proteins.

Melatonin in septic shock: Some recent concepts

Melatonin is a versatile molecule, synthesized not only in the pineal gland, but also in many other organs. Melatonin plays an important physiologic role in sleep and circadian rhythm regulation, immunoregulation, antioxidant and mitochondrial-protective functions, reproductive control, and regulation of mood. Melatonin has also been reported as effective in combating various bacterial and viral infections. Melatonin is an effective anti-inflammatory agent in various animal models of inflammation and sepsis, and its anti-inflammatory action has been attributed to inhibition of nitric oxide synthase with consequent reduction of peroxynitrite formation, to the stimulation of various antioxidant enzymes thus contributing to enhance the antioxidant defense, and to protective effects on mitochondrial function and in preventing apoptosis. In a number of animal models of septic shock, as well as in patients with septic disease, melatonin reportedly exerts beneficial effects to arrest cellular damage and multiorgan failure. The significance of these actions in septic shock and its potential usefulness in the treatment of multiorgan failure are discussed.

Effects of light cues on re-entrainment of the food-dominated peripheral clocks in mammals

Although the light/dark (LD) cycle is a weak Zeitgeber in peripheral clocks compared with food stimuli, whether the effect of light cues on the re-entrainment of peripheral clocks can be masked by that of the dominating food cue remains unknown. In the present study, the individual reversal of LD cycle for 7 days could not obviously affect circadian patterns of core clock genes (Bmal1, Cry1, Per1, and Dec1) in the liver and heart of restricted-fed rats. In contrast, reversing the feeding schedule together with the LD cycle markedly enhanced the re-entrainment of peripheral clocks compared with reversal of the feeding regimen alone. In addition, LD reversal alone for 7 days contrarily regulated the expression levels of Cry1, Per1, and Dec1 in the liver and heart. Moreover, daytime restricted feeding not only induced different phase shift rates of the four examined clock genes but also led to reversed phase shift direction in their resetting processes in these two peripheral clocks. Furthermore, the resetting sequences of these genes were also disparate between these two peripheral clocks. These observations suggest that the mechanisms underlying the liver and heart clocks are distinct, which may distinguish them from each other in the SCN-synchronized peripheral system.

Molecular Cloning and Circadian Regulation of Cryptochrome Genes in Japanese Quail (Coturnix coturnix japonica)

The circadian system is thought to have three components: input, pacemaker (internal clock), and output. Cryptochromes (Cry) are important clock genes, and recent findings indicate that these genes not only act as circadian photoreceptors but are also essential components in the negative feedback of the circadian system. As a first step toward understanding the avian circadian system, the authors tried to clone Japanese quail homologs of mammalian Crys and analyze their expression patterns in different circumstances. Partial cDNAs of qCry1 and qCry2, which are homologs of mammalian Cry1 and Cry2, respectively, were obtained and their gene expressions were analyzed. Both qCry1 and qCry2mRNAs were present in all the tissues examined. The oscillation patterns of the qCry1 transcripts were tissue specific and generally showed robust changes between daytime and nighttime; except for lung and testis tissues (which showed no detectable changes between daytime and nighttime), daytime levels were higher in all of the tissues examined. This rapid oscillation in qCry1 persisted through constant darkness or constant illumination, indicating that an endogenous clock controls these changes. In contrast, the expression of qCry2 did not oscillate in any tissue examined. In addition, in tissues of the pineal gland and eye, unexpected light exposure in the dark period was able to block the decrease in qCry1 transcripts or induce its expression. These findings, in conjunction with the established roles of CRYs in other species, led the authors to propose that in the circadian system, qCRYs may play important roles similar to the known roles of CRYs of other species, such as acting as circadian photoreceptors and as components of the circadian system.

Feeding-induced rapid resetting of the hepatic circadian clock is associated with acute induction of Per2 and Dec1 transcription in rats.

The synchronization of the master clock to photic cues is associated with a rapid induction of Per1, which plays an important role in initiating light-induced circadian resetting. However, the transcriptional mechanisms of clock gene expression in food-entrainable peripheral clocks have not been fully assessed. To understand how food cues might entrain a mammalian peripheral clock, we examined the responses in the expression of clock genes in rat livers to different feeding stimuli. The food-entrainable liver clock is more flexible than the light-entrainable SCN clock and can be reset rapidly at any time of day. A 30 min feeding stimulus was sufficient to significantly induce the expression of Per2 and Dec1 within 1 h and alter the transcript levels and circadian phases of other selected clock genes (Bmal1, Cry1, Per1, Per3, Dec2, and Rev-erba) in the liver clock at longer time intervals. Moreover, among the examined clock genes, Per2 was most sensitive to food cues, which could be significantly induced by a minimal amount of food. Furthermore, in contrast to the other hepatic clock genes, the feeding reversal-induced 12 h phase shift of Per2 could be rapidly and consistently accomplished, regardless of the shift of the light/dark cycle. In conclusion, the feeding-induced resetting of the circadian clock in the liver is associated with the acute induction of Per2 and Dec1 transcription, which may serve as the main and secondary input regulators that initiate this feeding-induced circadian resetting. (Author correspondence: azwfu2003@yahoo.com.cn)

Sardine peptide with angiotensin I-converting enzyme inhibitory activity improves glucose tolerance in stroke-prone spontaneously hypertensive rats.

An enzymatic hydrolysate of sardine protein (sardine peptide, SP) derived from sardine muscle possesses angiotensin I-converting enzyme (ACE) inhibitory activity. In the present study, we investigated the effect of SP on the blood glucose levels in stroke-prone spontaneously hypertensive rats (SHRSPs). Ten-week-old SHRSPs were assigned to three groups. The control group was given tap water for 4 weeks, while the experimental groups were given water containing SP (1 g/kg/d) or an ACE inhibitor, captopril (8 mg/kg/d). Treatment with SP and captopril decreased ACE activity in the kidney, aorta, and mesentery. There were no differences in fasting blood glucose levels among the three groups, whereas SP and captopril administration significantly suppressed the increase in blood glucose after glucose loading in the control SHRSPs. No difference was observed in plasma insulin levels among the three groups. Thus treatment with captopril and ACE-inhibitory sardine peptides ameliorated the glucose tolerance of this rat strain.

Melatonin in Bacterial and Viral Infections with Focus on Sepsis: A Review

Melatonin is a versatile molecule, synthesized not only by the pineal gland, but also in small amounts by many other organs like retina, gastrointestinal tract, thymus, bone marrow, lymphocytes etc. It plays an important role in various functions of the body like sleep and circadian rhythm regulation, immunoregulatory mechanism, free radical scavenger, antioxidant functions, oncostatic actions, control of reproductive functions, regulation of mood etc. Melatonin has also been found to be effective in combating various bacterial and viral infections. Its administration has been shown to be effective in controlling chlamydial infections, infections induced by Mycobacterium tuberculosis, and also in many viral infections. Molecular mechanisms of anti microbial actions of melatonin have suggested to be due to effects on free radical formation, direct regulation of duplication of bacteria, depletion of intracellular substrates like iron etc. Besides, it is effective in sepsis as demonstrated in various animal models of septic shock. Melatonins protective action against sepsis is suggested to be due to its antioxidant, immunomodulating and inhibitory actions against the production and activation of pro-inflammatory mediators. Use of melatonin has been beneficial in treating premature infants suffering from severe respiratory distress syndrome and septic shock. It has a potential therapeutic value in treating septic shock and associated multi organ failure in critically ill patients in addition to its antimicrobial and antiviral actions. The patents related to melatonins use for treatment of bacterial infections and its use in clinical disorders are included.

Differential Roles of Breakfast and Supper in Rats of a Daily Three-Meal Schedule Upon Circadian Regulation and Physiology

The timing of meals has been suggested to play an important role in circadian regulation and metabolic health. Three meals a day is a well-established human feeding habit, which in todays lifestyle may or may not be followed. The aim of this study was to test whether the absence of breakfast or supper significantly affects the circadian system and physiological function. The authors developed a rat model for their daily three meals study, whereby animals were divided into three groups (three meals, TM; no first meal, NF; no last meal, NL) all fed with the same amount of food every day. Rats in the NF group displayed significantly decreased levels of plasma triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and glucose in the activity phase, accompanied by delayed circadian phases of hepatic peripheral clock and downstream metabolic genes. Rats in the NL group showed lower concentration of plasma TC, HDL-C, and glucose in the rest phase, plus reduced adipose tissue accumulation and body weight gain. Real-time polymerase chain reaction (PCR) analysis indicated an attenuated rhythm in the food-entraining pathway, including down-regulated expression of the clock genes Per2, Bmal1, and Rev-erbα, which may further contribute to the delayed and decreased expression of FAS in lipogenesis in this group. Our findings are consistent with the conclusion that the daily first meal determines the circadian phasing of peripheral clocks, such as in the liver, whereas the daily last meal tightly couples to lipid metabolism and adipose tissue accumulation, which suggests differential physiological effects and function of the respective meal timings. (Author correspondence: azwfu2003@yahoo.com.cn)

Up-Regulation of Genes Related to the Ubiquitin-Proteasome System in the Brown Adipose Tissue of 24-h-Fasted Rats

The functional balance between brown adipose tissue (BAT) and white adipose tissue (WAT) is important for metabolic homeostasis. We compared the effects of fasting on the gene expression profiles in BAT, WAT and liver by using a DNA microarray analysis. Tissues were obtained from rats that had been fed or fasted for 24 h. Taking the false discovery rate into account, we extracted the top 1,000 genes that had been differentially expressed between the fed and fasted rats. In all three tissues, a Gene Ontology analysis revealed that the lipid and protein biosynthesis-related genes had been markedly down-regulated. The whole-body fuel shift from glucose to triacylglycerol and the induction of autophagy were also observed. There was marked up-regulation of genes in the ‘protein ubiquitination’ category particularly in BAT of the fasted rats, suggesting that the ubiquitin-proteasome system was involved in saving energy as an adaptation to food shortage.