入鹿山 容子

GPR109B (HM74) is a putative G protein-coupled receptor (GPCR) whose cognate ligands have yet to be characterized. GPR109B shows a high degree of sequence similarity to GPR109A, another GPCR that was identified as a high-affinity nicotinic acid (niacin) receptor. However, the affinity of nicotinic acid to GPR109B is very low. In this study, we found that certain aromatic D-amino acids, including D-phenylalanine, D-tryptophan, and the metabolite of the latter, D-kynurenine, decreased the activity of adenylate cyclase in cells transfected with GPR109B cDNA through activation of pertussis toxin (PTX)-sensitive G proteins. These D-amino acids also elicited a transient rise of intracellular Ca(2+) level in cells expressing GPR109B in a PTX-sensitive manner. In contrast, these D-amino acids did not show any effects on cells expressing GPR109A. We found that the GPR109B mRNA is abundantly expressed in human neutrophils. D-phenylalanine and D-tryptophan induced a transient increase of intracellular Ca(2+) level and a reduction of cAMP levels in human neutrophils. Furthermore, knockdown of GPR109B by RNA interference inhibited the D-amino acids-induced decrease of cellular cAMP levels in human neutrophils. These D-amino acids induced chemotactic activity of freshly prepared human neutrophils. We also found that D-phenylalanine and D-tryptophan induced chemotactic responses in Jurkat cells transfected with the GPR109B cDNA but not in mock-transfected Jurkat cells. These results suggest that these aromatic D-amino acids elicit a chemotactic response in human neutrophils via activation of GPR109B.

The spontaneously hypertensive rat (SHR) is a model of cardiomyopathy that displays a genetic defect in cardiac fatty acid (FA) translocase/CD36, a plasma membrane long-chain FA transporter. Therapy with medium-chain FAs, which do not require CD36-facilitated transport, has been shown to improve cardiac function and hypertrophy in SHRs despite persistent hypertension. However, little is known about the underlying molecular mechanisms. The aim of this study was to document the impact of medium-chain triglyceride (MCT) therapy in SHRs on the expression level and activity of metabolic enzymes and signaling pathways. Four-week-old male SHRs were administered MCT (SHR-MCT) or long-chain triglyceride (SHR-LCT) for 16 wk. We used Wistar-Kyoto (WKY) rats as controls (WKY-MCT and WKY-LCT). The SHR-MCT group displayed improved cardiac dysfunction [as assessed by left ventricular (LV) end-diastolic pressure and the positive and negative first derivatives of LV pressure/P value], a shift in the beta-myosin heavy chain (MHC)-to-alpha-MHC ratio, and cardiac hypertrophy compared with the SHR-LCT group without an effect on blood pressure. Administration of MCT of SHRs reversed the LCT-induced reduction in the cardiac FA metabolic enzymatic activities of long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) and medium-chain acyl-CoA dehydrogenase (MCAD). In the SHR-MCT group, the protein expression and transcriptional regulation of myocardial peroxisome proliferator-activated receptor-alpha, which regulates the transcription of LCHAD and MCAD genes, corresponded to the changes seen in those enzymatic activities. Furthermore, MCT intake caused an inhibition of JNK activation in SHR hearts. Collectively, the observed changes in the myocardial activity of metabolic enzymes and signaling pathways may contribute to the improved cardiac dysfunction and hypertrophy in SHRs following MCT therapy.

Background Primary polydipsia is a common complication in patients with chronic psychoes, particularly schizophrenia. Disease pathogenesis is poorly understood, but one contributory factor is thought to be dopamine dysregulation caused by prolonged treatment with neuroleptics. Both angiotensin-converting enzyme (ACE) and orexin (hypocretin) signaling can modulate drinking behavior through interactions with the dopaminergic system. Methods: We performed association studies on the insertion/deletion (I/D) sequence polymorphism of ACE and single nucleotide polymorpbisms within the prepro-orexin (HCRT), orexin receptor 1 (HCRTR1), and orexin receptor 2 (HCRM) genes. Genotypes were determined by polymerase chain reaction amplification, followed by either electrophoretic separation or direct sequencing. Results. The ACE I/D polymorphism showed no association with polydipsic schizophrenia. Screening of the orexin signaling system detected a 408 isoleucine to valine mutation in HCRTR1 that showed significant genotypic association with polydipsic-byponatremic schizophrenia (p = .012). The accumulation of this mutation was most pronounced in polydipsic versus nonpolydipsic schizophrenia (p = .0002 and p = .008, for the respective genotypic and allelic associations). The calcium mobilization properties and the protein localization of mutant HCRTR1 seem to be unaltered. Conclusion: Our preliminary data suggest that mutation carriers might have an increased susceptibility to polydipsia through an undetermined mechanism.

Myocardial Fiborosis and diastolic dysfunction in Deoxycorticosterone Acetate-Salt Hypertensive rats is ameliorated by the peroxisome proliferator-activated receptor-alpha activator fenofibrate, partly by suppressing inflammatory responses associated with the nuclear factor-kappa-B pathway.

Aims: Ageing impairs endothelial function such as the regulation of vascular tone. The release of nitric oxide (NO), which has a potent vasodilator effect and antiatherosclerotic property, is decreased in the aorta of aged rats. Exercise training, however, has been reported to increase the expression of endothelial NO synthase (eNOS) in the aorta of young rats. In aged rats, it is not known whether the expression of eNOS is altered by exercise training. We hypothesized that exercise training would improve the ageing-induced decrease in eNOS expression in vessels, and examined the messenger RNA (mRNA) and protein expression of eNOS in the aorta of sedentary-young rats (sedentary-young group; 4 months old), sedentary-aged rats (sedentary-aged group; 23 months old), and swim-trained aged rats (training-aged group; 23 months old, swimming training for 8 weeks, 5 days week(-1) , 90 min day(-1) ). Results: Body weight was significantly lower, and citrate synthase activity in the epitrochlearis muscle was significantly higher in the training-aged group compared with the sedentary-aged group. The mRNA expression of eNOS in the aorta was significantly higher in the training-aged group than in the sedentary-aged group, while it was significantly lower in both the sedentary-aged and training-aged groups than in the sedentary-young group. The expression of eNOS protein in the aorta was also significantly higher in the training-aged group than in the sedentary-aged group, while it was also significantly lower in the sedentary-aged group, but not in the training-aged group, than in the sedentary-young group. Conclusion: The present results revealed that the production of eNOS in the aorta decreases with ageing, and that the decreased production is increased by exercise training in aged rats, which may produce beneficial effects on the impaired cardiovascular system caused by ageing.

Peroxisome proliferator-activated receptor (PPAR)-alpha, a transcriptional activator, regulates genes of fatty acid (FA) metabolic enzymes. To study the contribution of PPAR-alpha to exercise training-induced improvement of FA metabolic capacity in the aged heart, we investigated whether PPAR-alpha signaling and expression of its target genes in the aged heart are affected by exercise training. We used hearts of sedentary young rat (4 mo old), sedentary aged rat (23 mo old), and swim-trained aged rat (23 mo old, training for 8 wk). The mRNA and protein expression of PPAR-alpha in the heart was significantly lower in the sedentary aged rats compared with the sedentary young rats and was significantly higher in the swim-trained aged rats compared with the sedentary aged rats. The activity of PPAR-alpha DNA binding to the transcriptional regulating region on the FA metabolic enzyme genes, the mRNA expression of 3-hydroxyacyl CoA dehydrogenase (HAD) and carnitine palmitoyl transferase-I, which are PPAR-alpha target genes, and the enzyme activity of HAD in the heart altered in association with changes of the myocardial PPAR-alpha mRNA and protein levels. These findings suggest that exercise training improves aging-induced down-regulation in myocardial PPAR-alpha-mediated molecular system, thereby contributing to the improvement of the FA metabolic enzyme activity in the trained-aged hearts.

The present study was designed to elucidate the role of p38 mitogen-activated protein kinase (p38) in the pathogenesis of inflammation, using a mouse contact hypersensitivity (CHS) model induced by 2,4-dinitro-1-fluorobenzene (DNFB). Ear swelling was induced by challenge with DNFB, accompanied by infiltration of mononuclear cells, neutrophils, and eosinophils and a marked increase in mRNA levels of cytokines such as interleukin (IL)-2, interferon (IFN)-gamma, IL-4, IL-5, IL-1beta, IL-18, and tumor necrosis factor-alpha in the challenged ear skin. Both ear swelling and the number of infiltrated cells in DNFB-challenged ear skin were significantly inhibited by treatment with SB202190, a p38 inhibitor. Furthermore, the DNFB-induced expression of all cytokines except IL-4 was significantly inhibited by treatment with SB202190. Ribonuclease protection assay revealed that the mRNA levels of chemokines such as IP-10 and MCP-1 in ear skin were markedly increased at 24 h after challenge with DNFB. The induction of these chemokines was significantly inhibited by treatment with SB202190. In p38alpha +/- mice, both ear swelling and infiltration of cells induced by DNFB were reduced compared with those in wild-type mice. However, induction of cytokines by DNFB was also observed in p38alpha +/- mice, although the induction of IFN-gamma, IL-5, and IL-18 was typically reduced compared with that in wild-type mice. Challenge with DNFB slightly induced IP-10 and MCP-1 mRNA in p38alpha +/- mice, with weaker signals than those in SB202190-treated wild-type mice. These results suggest that p38 plays a key role in CHS and is an important target for the treatment of CHS.

Background-Endothelin-1 (ET-1) is a potent endothelium-derived vasoconstrictor peptide. Exercise results in a significant redistribution of tissue blood flow, which greatly increases blood flow in active muscles but decreases it in the splanchnic circulation. We reported that exercise causes an increase of ET-1 production in the internal organ and then hypothesized that ET- 1 participates in the exercise-induced redistribution of tissue blood flow. We investigated the effects of acute endothelin-A (ETA)-receptor blockade on regional tissue blood flow during exercise in rats. Methods and Results-Regional blood flow in the kidney, spleen, stomach, intestine, and muscles was measured using the microsphere technique before and during treadmill running of 30 minutes duration at 30 m/min after pretreatment with either an ETA-receptor antagonist (TA-0201; 0.5 mg/kg) or vehicle in rats. Blood flow in the kidney, spleen, stomach, and intestine was decreased by exercise, but the magnitude of the decrease after pretreatment with TA-0201 was significantly smaller than that after pretreatment with vehicle. Furthermore, the increase in blood flow to active muscles induced by exercise was significantly smaller in rats pretreated with TA-0201 than those pretreated with vehicle. Conclusions-The present study revealed that ET-1-mediated vasoconstriction participates in the decrease of blood flow in the internal organs of rats during exercise, and therefore, that these actions of endogenous ET- 1 partly contribute to the increase of blood flow in active muscles during exercise. The data suggest that endogenous ET- 1 participates in the exercise-induced redistribution of tissue blood flow.

Vascular endothelial cells produce nitric oxide (NO), which is a potent vasodilator substance and has been proposed as having antiatherosclerotic property. Vascular endothelial cells also produce endothelin-1 (ET-1), which is a potent vasoconstrictor peptide and has potent proliferating activity on vascular smooth muscle cells. Therefore, ET-I has been implicated in the progression of atheromatous vascular disease. Because exercise training has been reported to produce an alteration in the function of vascular endothelial cells in animals, we hypothesized that exercise training influences the production of NO and ET-I in humans. The purpose of the present study was to examine whether chronic exercise could influence the plasma levels of NO (measured as the stable end product of NO, i.e., nitrite/nitrate [NOx]) and ET-I in humans. Eight healthy young subjects (20.3 +/- 0.5 yr old) participated in the study and exercised by cycling on a leg ergometer (70% VO2 max for 1 hour, 3-4 days/week) for 8 weeks. Venous plasma concentrations of NOx and ET-I were measured before and after (immediately before the end of 8-week exercise training) the exercise training, and also after the 4th and 8th week after the cessation of training. The VO2 max Significantly increased after exercise training. After the exercise training, the plasma concentration of NOx significantly increased (30.69 +/- 3.20 vs. 48.64 +/- 8.16 mu mol/L, p < 0.05), and the plasma concentration of ET-I significantly decreased (1.65 <plus/minus> 0.14 vs. 1.23 +/- 0.12 pg/mL, p < 0.05). The increase in NOx level and the decrease in ET-I level lasted to the 4th week after the cessation of exercise training and these levels (levels of NOx and ET-1) returned to the basal levels (the levels before the exercise training) in the 8th week after the cessation of exercise training. There was a significant negative correlation between plasma NOx concentration and plasma ET-I concentration. The present study suggests that chronic exercise causes an increase in production of NO and a decrease in production of ET-I in humans, which may produce beneficial effects (i.e., vasodilative and antiatherosclerotic) on the cardiovascular system. <(c)> 2001 Elsevier Science Inc. All rights reserved.

We investigated the role of xanthine oxidase-derived oxygen radicals in the development of endothelin-1-induced gastric: ulcer. Mucosal lipid peroxidation showed a peak 24 h after injection, while gastric mucosal haemodynamics were fully restored 26 h after endothelin-1 injection. Allopurinol and oxypurinol, but not superoxide dismutase or catalase, protected the gastric mucosa 24 h after endothelin-1 injection. Oxypurinol antagonized both the vasoconstrictor effect of endothelin-1 and the decrease in gastric ATP. All treatments on the second day after endothelin-1 injection significantly reduced gastric mucosal damage. Xanthine oxidase-derived oxygen radicals contributed largely to the exacerbation but they did not mediate the onset of endothelin-1-induced gastric ulcer. Pretreatment with probucol (500 mg/kg, p.o.) also protected the gastric mucosa from endothelin-1-induced mucosal injury by its antioxidant activity. Oxypurinol was gastroprotective through its vasoactive and energy saving actions. The haemodynamic background of endothelin-1-induced gastric ulcer consists of long lasting ischaemia and subsequent "reperfusion" which may be responsible for the late burst of oxygen radicals. (C) 2001 Elsevier Science B.V. All rights reserved.

It has been reported that probucol is a lipid-lowering agent having a strong antioxidative effect and inhibitory action on vascular smooth muscle cell proliferation. In this work, we studied the effect of treatment with a 1% probucol diet on pulmonary hypertension induced by monocrotaline (MCT) in rats. Rats were fed a control or 1% probucol-supplemented diet for 7 days, then given a single subcutaneous injection of 60 mg/kg MCT or saline, and continuously fed the same diet for 20 days, respectively. MCT caused an increase in right ventricular systolic pressure (RVSP), an indicator of pulmonary hypertension, and central venous pressure (CVP) on day 20. In rats receiving a diet containing 1% probucol, RVSP was significantly lower than that in rats treated with control diet, and CVP remained essentially at the basal level. On day 20, MCT also caused an increase in the ratio of right ventricular (RV) to body weight (BW), compared to the control value, indicating the development of RV hypertrophy in MCT rats. RV hypertrophy was significantly inhibited in 1% probucol-treated rats. These findings suggest that chronic treatment with probucol effectively inhibits the progression of pulmonary hypertension in rats. (C) 2000 Elsevier Science Inc. All rights reserved.

This study was designed to examine the immunomodulatory effects of dietary docosahexaenoic acid (DHP;) in the absence of eicosapentaenoic acid (EPA). We investigated the effects of feeding dietary DHA ethyl ester (DHA-Et) (97% pure) at levels of 4.8 wt% of the total diet and of feeding EPA ethyl ester (EPA-Et) (99% pure) at 4.8 wt% on the inflammatory response in the challenge phase of the contact hypersensitivity reaction (CHR) in the ears of mice sensitized with 2,4-dinitro-1-fluorobenzene (DNFB). The effect of DHA-Et on T lymphocytes at the CHR site was examined using anti-CD4 antibodies, Furthermore, we examined the cytokines formed at the CHR site on the mRNA level. It was found that 24 h after the challenge, DHA-Et but not EPA-Et reduced the ear swelling. Infiltration of inflammatory cells, in particular, CD4-positive T lymphocytes, into the ears in the challenge phase of CHR was observed. DHA-Et reduced the infiltration of CD4-positive T lymphocytes into the ears. DHA-Et also decreased the expression of interferon-li interleukin (IL)-6, IL-1 beta, and IL-2 mRNA in ears. These observations suggest that DHA, but not EPA, may exert an antiinflammatory and immunosuppressive effect. The immunosuppressive effectiveness of fish oil may be attributed mainly to DHA.

n-3系多価不飽和脂肪酸を含有した食用油脂であるしそ油, tuna oil及び鮭の魚卵リン脂質 (fish roe PL) はn-6脂肪酸を含有するとうもろこし油に比べて2, 4-dinitro-1-fluorobenzene (DNFB) で誘引された耳介の接触皮膚炎モデルの炎症反応を抑制する作用を持っている。Fish roe PLは耳介腫脹をもっとも強く抑制し, tuna oil及び漢方薬である柴朴湯がそれに続き, αリノレン酸を多量に含むしそ油はとうもろこし油に比べて抑制の傾向は見られたが有意な差は見られなかった。炎症性のCD4+T細胞の耳介への浸潤はfish roe PL投与によって抑制され, 耳介においてIFN-γ, IL-6, IL-1βのmRNAの発現抑制が観察された。本実験の結果でfish roe PLは遅延型のアレルギーを抑制する効果を持つことが示唆された。

n-3系多価不飽和脂肪酸を含有した食用油脂であるしそ油, tuna oil及び鮭の魚卵リン脂質 (fish roe PL) はn-6脂肪酸を含有するとうもろこし油に比べて2, 4-dinitro-1-fluorobenzene (DNFB) で誘引された耳介の接触皮膚炎モデルの炎症反応を抑制する作用を持っている。Fish roe PLは耳介腫脹をもっとも強く抑制し, tuna oil及び漢方薬である柴朴湯がそれに続き, αリノレン酸を多量に含むしそ油はとうもろこし油に比べて抑制の傾向は見られたが有意な差は見られなかった。炎症性のCD4+T細胞の耳介への浸潤はfish roe PL投与によって抑制され, 耳介においてIFN-γ, IL-6, IL-1βのmRNAの発現抑制が観察された。本実験の結果でfish roe PLは遅延型のアレルギーを抑制する効果を持つことが示唆された。

In isolated superior mesenteric arteries, vasodilation induced by calcitonin gene-related peptide (CGRP) was significantly larger in 12-week-old spontaneously hypertensive rats (SHR) than Wistar-Kyoto rats (WKY). In WKY and 6-week-old SHR, most of the vasodilator response to CGRP was abolished by N(w)-nitro-l-arginine and indomethacin. In contrast, the inhibitors caused no significant change in the response in 12-week-old SHR. Vasodilations induced by acetylcholine and isoproterenol were smaller in 12-week-old SHR than WKY, and that induced by sodium nitroprusside was comparable in both tissues. These results suggest that endothelium-independent vasodilator activity of CGRP is enhanced in hypertensive SHR, which overwhelms the decreased endothelium-dependent effects. Copyright (C) 1998 Elsevier Science B.V. All rights reserved.

The actions of protein S (PS) on the scratch injury-induced proliferation of rat astrocytes (AC) were studied. PS (10-300 nM) markedly inhibited [H-3]thymidine incorporation into injured AC. The effect of 100 nM PS was comparable with that of transforming growth factor-beta 1 (TGF-beta 1; 20 ng/ml). The incorporation of bromodeoxyuridine, which is usually detectable in AC along the border of the wound, was undetectable in the presence of 300 nM PS. The level of PS mRNA in the injured AC was slightly increased 15 h after the injury, although the level of its receptor, Tyro 3 mRNA was not changed significantly. The results of the present study suggest that PS plays an important role in tissue repair processes in the central nervous system (CNS) by suppressing the proliferation of AC as in the case of TGF-beta 1.

Although physiological processes related to vascular function differ greatly between resistance arteries and conduit arteries, it is not known whether the effects of endothelin-1 on these arteries differ in humans. In the present study, the conduit portion and the resistance portion of isolated human mesenteric arteries were suspended in a Krebs-Ringer solution. Norepinephrine and endothelin-1 produced concentration-dependent contractions in both portions. The EC(50) value of norepinephrine in the resistance portion (3.7 X 10(-7) M, n = 8) did not differ from that in the conduit portion (3.4 X 10(-7) M, n = 7). However, the EC(50) value of endothelin-1 in the resistance portion (3.0 x 10(-9) M, n = 8) was significantly lower than that in the conduit portion (1.1 X 10(-8) M, n = 7, P < 0.05). Although the maximum response to norepinephrine in the resistance portion (calculated as the percentage of 50 mM KCl-induced contraction) did not differ from that in the conduit portion, the maximum response to endothelin-1 in the resistance portion was significantly, greater than that in the conduit portion. These results indicate that endothelin-1 induces more potent constriction in resistance portion than in conduit portion in isolated human mesenteric arteries.

Although physiological processes related to vascular function differ greatly between resistance arteries and conduit arteries, it is not known whether the effects of calcitonin gene-related peptide (CGRP), a vasodilator neuropeptide, on these arteries differ in humans. In the present study, the conduit portion and the resistance portion of isolated human mesenteric arteries were suspended in a Krebs-Ringer solution. CGRP produced vasorelaxations in both portions. The EC(50) values were very low both in the resistance portion (2.3 X 10(-9) M, n = 7) and in the conduit portion (2.2 X 10(-9) M; n = 7). The maximum response to CGRP in the resistance portion Was significantly greater than that in the conduit portion (94.6 +/- 4.0% vs. 64.1 +/- 2.6% relaxation of methoxamine-induced precontraction, both n = 7, p < 0.01). These data suggest that CGRP is one of the most potent endogenous vasodilators in both the resistance portion and the conduit portion of the human mesenteric arteries, and that CGRP induces more potent vasorelaxation in the resistance portion than in the conduit portion of these arteries.

The regulatory systems of endogenous endothelin-1 (ET-1) production from intact tissues and the effects of produced ET-1 on vascular tonus by a closed circuit perfusion system of rat mesenteric artery were investigated. It was demonstrated that ET-1 is released from intact mesenteric arterial beds from Wistar rats both under basal conditions and after stimulation with arginine-vasopressin (AVP) (10(-10)-10(-9) M). Furthermore, AVP (10(-10)-10(-9) M) markedly and dose-dependently induced the expression of prepro ET-1 mRNA in the mesenteric arterial beds. Increased release of ET-1 by AVP may contribute to maintaining vascular tonus. Concomitant addition of actinomycin D inhibited the increased expression of prepro ET-1 mRNA and reduced the amount of immunoreactive ET-1 found during the 6-hour perfusate from AVP (10(-10) M)-stimulated and -unstimulated tissues.

Endothelin, a recently discovered endothelium-derived peptide, has been reported to produce potent vasoconstriction in various vessels of experimental animals. To study the involvement of endothelin in the regulation of vascular tonus in humans, isolated human mesenteric arteries were investigated by both pharmacological and immunohistochemical methods. The vasoconstrictor action of endothelin-1 was examined on ring segments of human mesenteric arteries. Endothelin-1 induced a slowly developing and sustained contraction, with an EC50 value (half-maximal effective concentration) of 2.9 x 10(-9) M, two orders of magnitude smaller than that of norepinephrine (EC50 of 3.9 x 10(-7) M), indicating that the vasoconstrictor action of endothelin-1 is about 100 times more potent than that of norepinephrine. The contractile effect of endothelin-1 was affected neither by adrenergic, cholinergic, histaminergic, nor serotonergic antagonists, nor by inhibitors of arachidonic acid metabolism. The vasoconstrictor response to endothelin-1 was effectively antagonized by nicardipine, a dihydropyridine Ca2+ channel blocker. Endothelin-1 profoundly augmented contractile response to Ca2+ in partially depolarized tissues. Immunohistochemical studies revealed for the first time that endothelin-like immunoreactivity was localized in endothelial cells of human mesenteric artery. The results of the present study indicate that endothelin-1 is one of the most potent vasoconstrictors in the human mesenteric artery and that it induces vasoconstriction via an ultimately accelerating Ca2+ influx through voltage-dependent Ca2+ channels. Since endothelin-1 can be located in human endothelial cells, it may play an important physiological or pathophysiological role.