Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
36 巻 , 1 号
選択された号の論文の28件中1~28を表示しています
Current Topics
  • Kouichi Tanonaka
    2013 年 36 巻 1 号 p. 1-
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
  • Iyuki Namekata, Yayoi Tsuneoka, Hikaru Tanaka
    2013 年 36 巻 1 号 p. 2-7
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    The pulmonary vein contains a myocardial layer extending from the left atrium, which is receiving attention as the source of ectopic electrical activity underlying atrial fibrillation. Electrophysiological and pharmacological analysis of the pulmonary vein myocardium performed in various experimental animal species have revealed characteristics such as presence of intracellular Ca2+ oscillations and low repolarizing potency. The automaticity of the pulmonary vein myocardium is affected by various neurotransmitters, hormones and pharmacological agents. Clarification of the mechanisms and regulation of pulmonary vein automaticity would lead to the development of novel therapeutic strategies and pharmacological agents for atrial fibrillation.
  • Junko Kurokawa, Tetsushi Furukawa
    2013 年 36 巻 1 号 p. 8-12
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    Gender differences play a major role in the manifestation of cardiovascular disease including cardiac arrhythmias. In particular, female sex is an independent risk factor for development of torsade de pointes (TdP) arrhythmias not only in congenital long QT syndromes but also in acquired long QT syndromes which occur as adverse effects of existing drugs. Recent clinical and experimental studies suggest that the gender differences may stem, at least in part, from gender differences in cardiac repolarization process, that is longer rate-corrected QT (QTC) interval in women than in men. In women, QTC interval and arrhythmic risks in TdP alter cyclically during menstrual cycle, suggesting a critical role of female sex hormones in cardiac repolarization process. These gender differences in fundamental cardiac electrophysiology result from variable ion channel expression and diverse sex hormonal regulation via long term genomic and acute non-genomic pathways, and sex differences in drug responses and metabolisms. In particular, non-genomic actions of testosterone and progesterone on cardiac ion channels likely to contribute to the gender differences in cardiac repolarization processes.
  • Yasufumi Katanasaka, Yoichi Sunagawa, Koji Hasegawa, Tatsuya Morimoto
    2013 年 36 巻 1 号 p. 13-17
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    Heart failure is one of the leading causes of death throughout the world. During the development and deterioration processes of heart failure, cardiomyocytes undergo maladaptive hypertrophy by altering hypertrophy-related gene expression. The zinc finger protein GATA4 is one of the transcription factors involved in the regulation of cardiomyocyte hypertrophy. In response to hypertrophic stimuli such as the synaptic nervous and rennin-angiotensin systems, GATA4 forms a large complex with various functional proteins including an intrinsic histone acetyltransferase, p300, and the disruption of this complex results in the inhibition of hypertrophic responses in cardiomyocytes. While such a transcriptional signal pathway is recognized as a critical event during cardiomyocyte hypertrophy, pharmacological heart failure therapy that targets this pathway has not been established. In order to develop novel heart failure therapy targeting the pathway in cardiomyocytes, we have studied the potential of curcumin, a p300 histone acetyltransferase inhibitor, as an agent for novel heart failure therapy. In this review, we describe a recent study on the cardiac transcriptional signal pathway, especially p300/GATA4 pathway, and a novel heart failure therapy using curcumin.
  • Atsushi Sanbe
    2013 年 36 巻 1 号 p. 18-22
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    Cardiomyopathies are defined as cardiac diseases of the myocardium with associated cardiac dysfunction. They are cardiac diseases in which heart muscle disease and/or measurable deterioration of cardiac muscle function occurs due to various causes, such as genetic and sporadic mutations of muscle proteins, as well as external factors such as hypertension, ischemia, and inflammation. In 1995, the WHO/International Society and Federation of Cardiology (ISFC) classified primary cardiomyopathy caused by intrinsic factors into five groups according to the dominant pathophysiology: dilated cardiomyopathy (DCM), hypertrophic cardiomyopathy (HCM), restricted cardiomyopathy (RCM), arrhythmogenic right ventricular cardiomyopathy (ARVC), and unclassified cardiomyopathy. Among these cardiomyopathies, DCM is the most prevalent and the most common reason for cardiac transplantation in adults and children. Many recent findings indicate that genetic and sporadic mutations of a number of muscle proteins, such as myofibrillar, structural, and Ca2+ regulating proteins, can cause DCM. In such cases, certain mutations often induce DCM with cardiac arrhythmia that is recognized as a potential trigger of sudden cardiac death. Thus, effective prognostic determination and appropriate cardiac care depend on accurate molecular and genetic diagnoses.
Regular Articles
  • Qi-Xiong Chen, Jing-Kun Miao, Chun Li, Xiao-Wen Li, Xiao-Mei Wu, Xiao- ...
    2013 年 36 巻 1 号 p. 23-30
    発行日: 2013/01/01
    公開日: 2013/01/08
    [早期公開] 公開日: 2012/10/18
    ジャーナル フリー
    For centuries, extracts of Acorus gramineus have been used extensively in traditional Chinese medicine for the treatment, management, and/or control of human ailments, including central nervous system disorders such as convulsions and epilepsy. In the present study, we investigated the anticonvulsant activity of chronic treatment with the plant’s major essential oil component (a-asarone, 50–200 mg/kg, per os (p.o.)) against maximal electroshock seizure (MES), pentylenetetrazole (PTZ)-induced seizures in mice, lithium–pilocarpine (LI–PILO)-induced status epilepticus (SE), and spontaneous recurrent seizures (SRSs) in rats and determined whether a single acute administration of a-asarone at various doses could produce anticonvulsant activity. As the standard antiepileptic drugs used, chronically administered a-asarone (50–200 mg/kg, p.o.) significantly delayed (p<0.05) the onset of, and antagonized maximal electroshock seizure and PTZ-induced seizures. Chronically administered a-asarone (50–200 mg/kg) also profoundly antagonized LI–PILO-induced seizures. The SE incidence, SE latency and seizure severity as well as mortality were significantly reduced after treatment with a-asarone at different doses. Higher doses of a-asarone (100–200 mg/kg) significantly reduced spontaneous recurrent seizure incidence, severity, and seizure frequency during treatment in LI–PILO-induced SRSs rats. On the other hand, a single acute administration of a-asarone (50–200 mg/kg) produced weak anticonvulsant activity in MES and PTZ-induced seizures. The results of this laboratory animal study indicate that chronically administered a-asarone possesses anticonvulsant activity in the mammalian experimental model used, and thus suggest that a-asarone may be used as a natural supplementary remedy in the management of convulsions and epilepsy.
  • Motoki Bito, Takashi Tomita, Mika Komori, Takanori Taogoshi, Yasuhiro ...
    2013 年 36 巻 1 号 p. 31-35
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    Fluoroquinolones reportedly induce hypoglycemia through stimulation of insulin secretion from pancreatic β-cells via inhibition of KATP channels and activation of L-type voltage-dependent Ca2+ channels. In physiological condition, the cytosolic Ca2+ concentration ([Ca2+]c) is also regulated by release of Ca2+ from intracellular Ca2+ stores. In this study, we investigated the mechanism of insulin secretion induced by fluoroquinolones, with respect to intracellular Ca2+ stores. Even where the absence of supplemental extracellular Ca2+, insulin secretion and [Ca2+]c were increased by gatifloxacin, levofloxacin or tolbutamide. Insulin secretion and the rise of [Ca2+]c induced by fluoroquinolones were reduced by depleting of Ca2+ in endoplasmic reticumum (ER) by thapsigargin, and inhibiting ryanodine receptor of ER by dantrolene. Inhibition of inositol 1,4,5-triphosphate receptor of ER by xestospongin C suppressed insulin secretion induced by fluoroquinolones, whereas it did not affect [Ca2+]c. Destruction of acidic Ca2+ stores such as lysosome and lysosome-related organelles by glycyl-L-phenylalanine-2-nephthylamide (GPN) did not affect insulin secretion and the rise of [Ca2+]c induced by fluoroquinolones. The increase in insulin and [Ca2+]c induced by tolbutamide were reduced by thapsigargin, dantrolene, and GPN but not by xestospongin C. In conclusion, fluoroquinolones induces Ca2+ release from ER mediated by the ryanodine receptor, and the reaction might involve in insulin secretion. Sulfonylureas induce Ca2+ release from GPN-sensitive acidic Ca2+ stores, but fluoroquinolones did not.
  • Masaki Okawara, Yoshihiro Tokudome, Hiroaki Todo, Kenji Sugibayashi, F ...
    2013 年 36 巻 1 号 p. 36-40
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    Orally administrated diosgenin, a steroidal saponin found in several plants including Dioscorea villosa, recovers skin thickness reduced in ovariectomized mice, and plays an important role in the treatment of hyperlipidemia. Thus, diosgenin is an active element of cosmeceutical and dietary supplements. However, we have already elucidated that the skin distribution and absolute oral bioavailability of diosgenin is very low. The aim of this study is to evaluate the efficacy of diosgenin–cyclodextrin (CD) complexes in improving the skin concentration of diosgenin. The formation of the CD complex was indicated by powder X-ray diffraction (XRD), differential scanning calorimetry (DSC), and scanning electron microscope (SEM) studies. Oral administration of the diosgenin/β-CD complex resulted in a significant enhancement in terms of the skin distribution of diosgenin, maximum plasma level (Cmax), area under the plasma concentration–time curve (AUC), and absolute oral bioavailability over those of the drug alone. These results suggest that the inclusion complex of diosgenin/β-CD can be used to improve low skin content of diosgenin.
  • Takahiko Ono, Kohei Kamikado, Tatsuya Morimoto
    2013 年 36 巻 1 号 p. 41-47
    発行日: 2013/01/01
    公開日: 2013/01/08
    [早期公開] 公開日: 2012/11/07
    ジャーナル フリー
    Oxidative stress and peritubular capillary (PTC) injury are involved in the progression of chronic kidney disease (CKD). We investigated protective effects of Shichimotsu-koka-To (SKT), a Japanese traditional Kampo prescription, against nephrosclerosis and hypertension on a CKD model due to irreversible nephritis. Six-week-old male Wistar rats were subjected to uninephrectomy, and to injection of rabbit anti-thymocyte serum. SKT treatment was continued for 15 weeks, blood pressure was measured, and then renal specimens were collected. PTC networks were detected by immunostaining for CD-31. And superoxide dismutase (SOD)-like activity in the tissue was evaluated. Blood pressure in the SKT group, as well as sham group, was significantly lower than with the vehicle. SKT markedly ameliorated renal function, which was evaluated with urea nitrogen clearance. Compared with the vehicle, SKT treatment lowered both the glomerular enlargement and hyper-cellularity by 80%, and decreased the extracellular matrix area by 75%. SKT treatment also suppressed tubular injury, and maintained PTC networks. Furthermore, SKT recovered SOD-like activity to the basal levels. These results suggest that SKT may be useful for the treatment of CKD during the progression to nephrosclerosis, through the mechanisms of anti-oxidative activity and maintenance of PTC networks.
  • Kosuke Endo, Yukiko Naito, Xu Ji, Michio Nakanishi, Teruo Noguchi, Yoi ...
    2013 年 36 巻 1 号 p. 48-54
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    MicroRNAs (miRNAs) are endogenous small RNAs that are 18–23 nucleotides long. Recently, plasma miRNAs were reported to be sensitive and specific biomarkers of various pathological conditions. In the present study, we focused on miR-210, which is known to be induced by hypoxia and might therefore be an excellent biomarker for congestive heart failure. Plasma miR-210 levels and expression levels in mononuclear cells and skeletal muscles were elevated in Dahl salt-sensitive rats with heart failure. We also assessed miR-210 expression in patients with heart failure. The miR-210 expression levels in the mononuclear cells of patients with NYHA III and IV heart failure according to the New York Heart Association (NYHA) functional classification system were significantly higher than those with NYHA II heart failure and controls. Although no significant correlation was observed between plasma brain natriuretic peptide (BNP) and plasma miR-210 levels in patients with NYHA II heart failure, patients with an improved BNP profile at the subsequent hospital visit were classified in a subgroup of patients with low plasma miR-210 levels. Plasma miR-210 levels may reflect a mismatch between the pump function of the heart and oxygen demand in the peripheral tissues, and be a new biomarker for chronic heart failure in addition to plasma BNP concentrations.
  • Young Mi Ha, Hye Jin Lee, Daeui Park, Hyoung Oh Jeong, Ji Young Park, ...
    2013 年 36 巻 1 号 p. 55-65
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    We simulated the docking of the tertiary structure of mushroom tyrosinase with our compounds. From the structure-tyrosinase inhibitory activity relationship, it is notable that compounds 4, 8 and 11 showed similar or better activity rates than kojic acid which was used as a positive control. Compounds 17, 21, and 23 among benzene analogs that possess the same substituent showed significantly lower tyrosinase inhibitory effects. Therefore, we have confirmed that among the compounds showing better tyrosinase inhibitory effects than kojic acid, the compounds with triene analogs have better tyrosinase inhibitory effect than the compounds with benzene analogs. Docking simulation suggested the mechanism of compounds by several key residues which had possible hydrogen bonding interactions. The pharmacophore model underlined the features of active compounds, 4,4′-((1E,3E,5E)-hexa-1,3,5-triene-1,6-diyl)diphenol, 5,5′-((1E,3E,5E)-hexa-1,3,5-triene-1,6-diyl)bis(2-methoxy-phenol), and 5,5′-((1E,3E,5E)-hexa-1,3,5-triene-1,6-diyl)dibenzene-1,3-diol among triene derivatives which had several hydrogen bond groups on both terminal rings. The soundness of the docking results and the agreement with the pharmacophores suggest that it can be conveniently exploited to design inhibitors with an improved affinity for tyrosinase.
  • Yuko Ogawa, Yoshitaka Taketomi, Makoto Murakami, Masafumi Tsujimoto, R ...
    2013 年 36 巻 1 号 p. 66-75
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    Small non-coding RNAs, such as microRNAs (miRNAs), are involved in diverse processes, including organ development and tissue differentiation. Exosomes are small membrane vesicles (30–100 nm in diameter) produced by numerous cells. Recently, exosomes have been shown to contain miRNAs. However, the small RNAs contained in exosomes are not fully characterized. In a previous study, we found at least two types of salivary exosome that are different in size and have different proteomes. Studies of salivary exosomal small RNAs are limited to miRNAs. In this study, we examined small RNA transcriptomes using next generation sequencing technology to elucidate a full transcriptome set of small RNAs expressed in the two types of salivary exosomes and in whole saliva (WS). Many types of small RNA, such as miRNA, piwi-interacting RNA (piRNA), small nucleolar RNA (snoRNA) and other small RNAs are contained in salivary exosomes and WS. Among these small RNAs we identified novel miRNA candidates.
  • Takako Yoshino-Furukawa, Yasue Maeda, Aya Kikuchi, Hiroyuki Sakuma, Ka ...
    2013 年 36 巻 1 号 p. 76-81
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    The pharmacological properties of the novel neurokinin-1 (NK1) receptor antagonist FK886, ([3,5-bis(trifluoromethyl)phenyl][(2R)-2-(3-hydroxy-4-methylbenzyl)-4-{2-[(2S)-2-(methoxymethyl)morpholin-4-yl]ethyl}piperazin-1-yl]methanone dihydrochloride), were studied. FK886 potently inhibited the binding of [125I]Bolton-Hunter-labeled substance P ([125I]BH-SP; 100 pM) to human NK1 receptors expressed in Chinese hamster ovary (CHO) cells (IC50=0.70 nM). It also possessed high affinities for dog, ferret, gerbil and guinea pig NK1 receptors, but not for rat NK1 receptor. FK886 was highly selective for the NK1 receptor, with 250- and >20000-fold selectivity for human NK1 over NK2 and NK3, respectively. Further, it did not inhibit radioligand binding at 54 different sites, including receptors, ion channels and transporters. FK886 inhibited substance P (3.2 nM)-induced inositol phosphate formation in human NK1 receptor-expressing CHO cells (IC50=1.4 nM) without stimulating NK1 receptors. The antagonism exerted by FK886 against human NK1 receptor was insurmountable in saturation binding experiments, with both the affinity and Bmax of [125I]BH-SP being significantly reduced. After intravenous administration, FK886 (0.01–0.1 mg/kg) dose-dependently inhibited the foot-tapping behavior induced by intracerebroventricular administration of a selective NK1 receptor agonist, GR73632 (10 pmol), in gerbils, with significant inhibition being observed at doses of 0.032–0.1 mg/kg, indicating excellent brain penetration. The brain penetration of FK886 was further demonstrated by the cerebral distribution of radioactivity after intravenous injection of radiolabeled FK886. Taken together, these results demonstrate that FK886 is a potent, highly selective and centrally active, insurmountable antagonist of the NK1 receptor, and suggest that FK886 antagonizes various NK1 receptor-mediated biological effects in the central nervous system.
  • Sa Rang Oh, Su-Jin Kim, Dong Hyun Kim, Jong Hoon Ryu, Eun-Mi Ahn, Ji W ...
    2013 年 36 巻 1 号 p. 82-88
    発行日: 2013/01/01
    公開日: 2013/01/08
    [早期公開] 公開日: 2012/11/05
    ジャーナル フリー
    Memory impairment is the most common symptom in patients with Alzheimer’s disease (AD). Angelica keiskei (AK) has traditionally been used as a diuretic, laxative, analeptic and galactagogue. However, the anti-amnesic effects of AK and its molecular mechanisms have yet to be clearly elucidated. The aim of the present study is to evaluate the effects of AK on scopolamine-induced memory impairments in mice. The regulatory effect of AK on memory impairment was investigated using passive avoidance, Y-maze and the Morris water maze tasks. Acetylcholinesterase (AChE) activity assay was performed to investigate the cholinergic antagonistic effect of AK in the hippocampus. The effect of AK on phosphorylation of cAMP response element-binding protein (CREB) and expression of brain-derived neurotrophic factor (BDNF) were evaluated by Western blot assays and immunohistochemistry. The findings showed that AK significantly attenuated scopolamine-induced cognitive impairment in mice. Increase of AChE activity caused by scopolamine was significantly attenuated by AK. Additionally, AK significantly recovered the phosphorylation of CREB and expression of BDNF reduced by scopolamine in the hippocampus. Taken together, these results provide experimental evidence that AK might be a useful agent in preventing deficit of learning and memory caused by AD and aging.
  • Junichi Iida, Toshiyuki Kudo, Kento Shimada, Yoshiyuki Yatsuno, Saori ...
    2013 年 36 巻 1 号 p. 89-95
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    Metronidazole (MTZ) ointment has been used widely as a hospital preparation against cancerous malodor. Although cancerous tissue with ulcer-like symptoms is likely to have a higher capacity to absorb drugs than normal skin, the extent to which MTZ is absorbed when a topical preparation is applied to cancerous tissue remains unclear. Furthermore, few studies have investigated the drug interactions involving MTZ despite its long use in clinical practice. In the present study, plasma concentration of MTZ was measured in a breast cancer patient using MTZ ointment for cancerous malodor and basic research was also conducted with the objective of investigating the safety of topical MTZ from a pharmacokinetic perspective. 4.75 µg/mL (27.8 µM) of MTZ was detected in the patient’s plasma, which was close to the plasma concentration after oral dosage of MTZ. In a metabolic inhibition study using human liver microsomes, cytochrome P450 (CYP) 2C9-mediated hydroxylation of S-warfarin was almost unaffected by MTZ at the corresponding concentrations. In addition, 3-d repeated oral administration of MTZ (200 mg/kg/d) to rats did not show any significant effects on the hepatic mRNA levels of various CYP isozymes and CYP2C protein levels. These results suggest that the reported interaction of oral MTZ and S-warfarin was not due to CYP2C9 inhibition and that drug interactions via inhibition of CYP2C9 is unlikely to occur when MTZ ointment is applied to ulcerous skin. This information should be valuable for assessing the safety of MTZ ointment used for mitigating cancerous malodor.
  • Tomoaki Kurosaki, Masafumi Uematsu, Kenichiro Shimoda, Kiyoshi Suzuma, ...
    2013 年 36 巻 1 号 p. 96-101
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    In this experiment, we developed anionic ternary complexes for effective and safe ocular gene delivery. Ternary complexes were constructed by coating plasmid DNA (pDNA)/polyethylenimine (PEI) complex with anionic polymers such as γ-polyglutamic acid (γ-PGA) and chondroitin sulfate (CS). The cationic pDNA/PEI complex showed high gene expression on the human retinal pigment epithelial cell line, ARPE-19 cells. The pDNA/PEI complexes, however, also showed high cytotoxicity on the cells and aggregated strongly in the vitreous body. On the other hand, the anionic ternary complexes showed high gene expression on ARPE-19 cells without such cytotoxicity and aggregation. After intravitreous administration of the complexes, the anionic ternary complexes showed high gene expression in the retina. These results strongly indicate that anionic ternary complexes are suitable for effective and safe ocular gene therapy.
Notes
  • Wan Gu, Kyung-Ah Kim, Dong-Hyun Kim
    2013 年 36 巻 1 号 p. 102-107
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    Ginseng (the root of Panax ginseng C. A. MEYER), which contains protopanaxadiols and protopanaxatriols as its main constituents, has been used for many disorders, such as cancer, diabetes, inflammation, and hyperlipidemia. Of these ginsenosides, protopanaxadiol ginsenoside Rh2 alone is reported to inhibit adipogenesis in 3T3-L1 in vitro. Therefore, we investigated the effect of protopanaxatriol ginsenoside Rh1 on adipogenesis in 3T3-L1 cells and high fat diet-induced obesity (DIO) mice. Treatment with ginsenoside Rh1 inhibited adipogenesis, as evidenced by Oil red O staining and lipid droplet extraction assay. Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed that ginsenoside Rh1 decreased the expressions of peroxisome proliferator-activated receptor (PPAR)-γ, CCAAT/enhancer-binding protein (C/EBP)-α, fatty acid synthase, and adipocyte fatty acid-binding protein. Oral administration of ginsenoside Rh1 (20 mg/kg) suppressed body and epididymal fat weight gains and plasma triglyceride level in DIO mice. Ginsenoside Rh1 also inhibited the expressions of PPAR-γ, C/EBP-α, fatty acid synthase, adipocyte fatty acid-binding protein, as well as F4/80, CD68, tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β in DIO mice by real time PCR analysis. Based on these findings, ginsenoside Rh1 may ameliorate obesity, by inhibiting adipocyte differentiation and inflammation.
  • Sin Jee Son, Ki-Jong Rhee, Jaewon Lim, Tae Ue Kim, Tack-Joong Kim, Yoo ...
    2013 年 36 巻 1 号 p. 108-113
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    Triglyceride (TG) induces macrophage cell death which contributes to the development of atherosclerosis. We confirmed that exogenous TG accumulates in human THP-1 macrophages and causes cell death. TG treated THP-1 macrophages exhibited no change in tumor necrosis factor (TNF)-α, interleukin (IL)-18, macrophage inflammatory protein (MIP)-1α, and IL-1R1 receptor mRNA expression. However, there was a marked decrease in IL-1β mRNA expression but an increase in IL-1β protein secretion. Decreased expression of IL-1β mRNA and increased secretion of IL-1β protein was not the direct cause of cell death. Until now, TG was assumed to induce necrotic cell death in macrophages. Since caspase-1 is known to be involved in activation and secretion of IL-1β protein and pyroptotic cell death, next we determined whether caspase-1 is associated with TG-induced macrophage cell death. We found an increase in caspase-1 activity in TG-treated THP-1 macrophages and inhibition of caspase-1 activity using a specific inhibitor partially rescued cell death. These results suggest activation of the pyroptotic pathway by TG. This is the first report implicating the activation of caspase-1 and the triggering of the pyroptosis pathway in TG-induced macrophage cell death.
  • Takako Yoshihara, Yuzuru Yonoki, Maki Saito, Tsutomu Nakahara, Kenji S ...
    2013 年 36 巻 1 号 p. 114-119
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    β1- and β2-Adrenergic receptors (β1-AR and β2-AR) are co-expressed in numerous tissues, for example, heart and bladder. They play a very important role in the responses of a variety of organs to sympathetic nerve stimulation. Recent studies suggest that many G protein-coupled receptors, such as β1-AR, β2-AR, μ opioid receptor and δ opioid receptor, can form homo- and heterooligomers. Previous studies demonstrated that the β1-AR and β2-AR formed dimers in living HEK 293 cells. The aim of the present study is to investigate whether such heterooligomerization affect the agonist-induced receptor internalization in the CHO-K1 cells stably co-expressing β1-AR and β2-AR. Using co-immunoprecipitation, we confirmed that β1-AR and β2-AR formed heterooligomers in the CHO-K1 cells. In cells co-expressing β1-AR and β2-AR, 30% of β1-AR was internalized by isoproterenol, whereas only 20% of β1-AR was internalized in cells expressing the β1-AR alone. Heterooligomerization did not affect the ratio of internalized β2-AR. Salmeterol, a specific β2-AR agonist, broke β1-AR/β2-AR heterooligomers, and induced β2-AR-specific internalization in cells co-expressing β1-AR and β2-AR. The present study demonstrated that heterooligomerization between β1-AR and β2-AR accelerates the isoproterenol-promoted internalization of the β1-AR, and that salmeterol induces β2-AR-specific internalization in Chinese hamster ovary (CHO) cells stably co-expressing β1-AR and β2-AR.
  • Zehua Bian, Norihiko Furuya, Dong-Mei Zheng, Juan Alejandro Oliva Trej ...
    2013 年 36 巻 1 号 p. 120-124
    発行日: 2013/01/01
    公開日: 2013/01/08
    [早期公開] 公開日: 2012/10/30
    ジャーナル フリー
    Ferulic acid (FA), a naturally occurring polyphenol abundant in vegetables and rice bran, is known to possess a potent antioxidant activity, thereby protecting cells from oxidative stress. In the present study, we show that in addition to its known anti-oxidant activity, ferulic acid exerts substantial inhibitory activity on cellular mammalian target of rapamycin (mTor)-signaling pathways. In HeLa cells and mouse primary hepatocytes cultured with conventional nutrient-rich media, ferulic acid (1 mM) elicited dephosphorylation of S6 kinase and its substrate ribosomal S6. The dephosphorylating activity of ferulic acid was almost comparable to that of rapamycin, an established mTor inhibitor (TORC1). We next investigated the effect of ferulic acid on autophagy, a major cellular degradative process, which significantly contributes to the maintenance of cell homeostasis. Using a conventional green fluorescent protein-microtubule-associated protein IA/IB light chain 3 (GFP-LC3) dot assay to evaluate autophagy flux, we showed that ferulic acid caused a significant increase in GFP-LC3 dots under serum-rich conditions in HeLa cells. The enhancement of autophagic flux by ferulic acid was almost equivalent to that of rapamycin. Furthermore, ferulic acid significantly enhanced autophagic degradation of 14C-leucine-labeled long-lived proteins of cultured mouse hepatocytes under nutrient-rich conditions, but not nutrient-deprived conditions. These results indicate that ferulic acid is almost the equivalent of rapamycin in the ability to inhibit mTor (TORC1), which makes it a potent activator of basal autophagy.
  • Hyung Sun Lim, Jae-Min Kim, Jae-Gyun Choi, Young Kwon Ko, Yong Sup Shi ...
    2013 年 36 巻 1 号 p. 125-130
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    Peripheral or central nerve injury often leads to neuropathic pain. Although ketamine and pregabalin are first line options for the treatment of neuropathic pain, their clinical application is limited due to side effects such as sedation, dizziness and somnolence. We designed this study to determine whether the intrathecal (i.t.) co-treatment with ketamine and pregabalin at sub-effective low doses would elicit a sufficient pain relief without producing side effect in a neuropathic pain mouse model. At day 7 after chronic constriction injury (CCI) of sciatic nerve, dose dependent effects of i.t. ketamine (3, 10, 30, 100 µg) or i.t. pregabalin (10, 30, 100 µg) on mechanical allodynia and thermal hyperalgesia were measured. For combination treatment, 3 or 10 µg of ketamine and 30 µg of pregabalin were selected because these doses of drugs were not effective on neuropathic pain. Interestingly, combined i.t. treatment groups (ketamine 3 µg+pregabalin 30 µg and ketamine 10 µg+pregabalin 30 µg) produced strong analgesia on neuropathic pain although these doses of ketamine and pregabalin alone are not effective. Moreover, rota rod test revealed that normal motor function was not affected by combined treatment while i.t. ketamine at doses above 10 µg showed a significant motor dysfunction. Results of this study suggested that i.t. co-treatment with ketamine and pregabalin at sub-effect low doses may be a useful therapeutic method for the treatment of neuropathic pain patients.
  • Reona Takabatake, Mari Onishi, Tomohiro Koiwa, Satoshi Futo, Yasutaka ...
    2013 年 36 巻 1 号 p. 131-134
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    A novel real-time polymerase chain reaction (PCR)-based quantitative screening method was developed for three genetically modified soybeans: RRS, A2704-12, and MON89788. The 35S promoter (P35S) of cauliflower mosaic virus is introduced into RRS and A2704-12 but not MON89788. We then designed a screening method comprised of the combination of the quantification of P35S and the event-specific quantification of MON89788. The conversion factor (Cf) required to convert the amount of a genetically modified organism (GMO) from a copy number ratio to a weight ratio was determined experimentally. The trueness and precision were evaluated as the bias and reproducibility of relative standard deviation (RSDR), respectively. The determined RSDR values for the method were less than 25% for both targets. We consider that the developed method would be suitable for the simple detection and approximate quantification of GMO.
  • Takashi Kikuchi, Xin Pan, Koichi Ishii, Yasuhiro Nakamura, Eri Ogihara ...
    2013 年 36 巻 1 号 p. 135-139
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    12-O-Acetylazedarachin B (1), isolated from the fruit extract of Melia azedarach, exhibited potent cytotoxicity against leukemia (HL-60) (IC50 0.016 µM) and stomach (AZ521) (IC50 0.035 µM) cancer cell lines. Upon assessing the apoptosis-inducing activity in HL-60 cells, compound 1 exhibited induction of apoptosis detected by the observation of membrane phospholipid exposure and DNA fragmentation in flow cytometry. Western blot analysis showed that 1 markedly reduced the levels of procaspases-3, 8, and 9, while being increased the levels of cleaved caspases-3, 8, and 9. In addition, compound 1 increased significantly Bax/Bcl-2 ratio. These results suggested that 1 induced apoptotic cell death in HL-60 via both mitochondrial and death receptor-mediated pathways. Therefore, compound 1 may be promising lead compound for developing an effective drug for treatment of leukemia. Flow cytometric analysis suggested that the cytotoxicity of 1 against AZ521 is due to inducing apoptosis as well as necrosis with the latter predominated.
  • Ji-Jing Yan, Jun-Sub Jung, Taek-Keun Kim, Md. Ashraful Hasan, Chang-Wo ...
    2013 年 36 巻 1 号 p. 140-143
    発行日: 2013/01/01
    公開日: 2013/01/08
    [早期公開] 公開日: 2012/10/16
    ジャーナル フリー
    We previously reported the protective effects of long-term administration of ferulic acid against the in vivo toxicity of β-amyloid peptide administered intracerebroventricularly in mice. In the present study, we investigated the effects of ferulic acid in transgenic amyloid precursor protein (APP)swe/presenilin 1 (PS1)dE9 (APP/PS1) mouse model of Alzheimer disease (AD). Chronic (for 6 months from the age of 6 to 12 months) oral administration of ferulic acid at a dose of 5.3 mg/kg/day significantly enhanced the performance in novel-object recognition task, and reduced amyloid deposition and interleukin-1 beta (IL-1β) levels in the frontal cortex. These results suggest that ferulic acid at a certain dosage could be useful for prevention and treatment of AD.
  • Yoshiharu Suzuki, Masato Homma, Masato Abei, Ichinosuke Hyodo, Yukinao ...
    2013 年 36 巻 1 号 p. 144-146
    発行日: 2013/01/01
    公開日: 2013/01/08
    [早期公開] 公開日: 2012/10/25
    ジャーナル フリー
    The effects of solute carrier family 29 member 1 (SLC29A1) single nucleotide polymorphism (SNP), rs6932345 and rs747199, on SLC29A1 mRNA expression were examined. The expression levels of SLC29A1 mRNA in peripheral blood mononuclear cells (PBMCs) isolated from 46 healthy subjects (28 males and 18 females) was compared between wild-type and mutant carriers. The mRNA levels in the rs6932345 wild-type (AA genotype) was 1.71 times that in the mutation carriers (AC/CC genotype) (p<0.05). Similar results were observed for rs747199, because rs747199 was linked with rs6932345 at a frequency of 84.8%. It was confirmed that wild-type for rs6932345 and rs747199 showed higher SLC29A1 mRNA expression in PBMCs.
  • Minoru Sugiura, Kazunori Ogawa, Masamichi Yano
    2013 年 36 巻 1 号 p. 147-151
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    Fruits and vegetables contain numerous antioxidants, such as carotenoids. Recent epidemiologic studies have demonstrated that a high dietary consumption of fruit and vegetables rich in carotenoids or with high serum carotenoid concentrations results in lower risks of certain cancers, diabetes, and cardiovascular disease. These results indicate that absorbed carotenoids are stored in various organs. Previously, we found that β-cryptoxanthin, found especially in Satsuma mandarin (Citrus unshiu MARC.), is easily absorbed and can also survive for a relatively long time in the human body; however, little is known about the absorption, storage, and tissue distribution of β-cryptoxanthin. In this study, we measured serum and the content of β-cryptoxanthin in several rat tissues after chronic ingestion of Satsuma mandarin extract rich in β-cryptoxanthin. Rats were fed a standard commercial diet containing Satsuma mandarin extract (containing β-cryptoxanthin at 11.7 mg/kg diet) for eight weeks. After 3 h of fasting, serum, liver, spleen, kidney, lung, heart, testis, brain, and epididymal fat were collected. The concentrations of β-cryptoxanthin in serum and tissues were evaluated by high-performance liquid chromatography. There was a wide range in the tissue levels of β-cryptoxanthin; liver had the greatest value, with 1265.3 ng/g tissue, followed by spleen, kidney, lung, heart, brain, and testis. Epididymal fat had the lowest value, with 6.99 ng/g tissue. β-Cryptoxanthin was also detected in serum in a concentration of 5.76 ng/mL. These results indicate that β-cryptoxanthin is easily absorbed and accumulated in several organs.
  • Dami Jeong, Kousuke Watari, Takayuki Shirouzu, Mayumi Ono, Keiichi Koi ...
    2013 年 36 巻 1 号 p. 152-157
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    Metastasis occurs when cancer cells detach from a tumor, travel to distant sites in the body and develop into tumors in these new locations. Most cancer patients die from metastases. Among the various forms of cancer metastasis, lymphatic metastasis is an important determinant in cancer treatment and staging. In this study, we investigated lymphangiogenesis inhibitors from crude drugs used in Japan and Korea. The three crude drugs Saussureae Radix, Psoraleae Semen and Aurantti Fructus Immaturus significantly inhibited the proliferation of temperature-sensitive rat lymphatic endothelial (TR-LE) cells in vitro. By a chromatographic method using bioassay-guided fractionation methods, costunolide (1) and dehydrocostus lactone (2) from S. Radix, p-hydroxybenzaldehyde (3), psoralen (4), angelicin (5), psoracorylifol D (6), isobavachalone (7), bavachinin (8) Δ3,2-hydroxybakuchiol (9) and bakuchiol (10) from P. Semen and cis-octadecyl ferulate (11), (2R)-3β,7,4′-trihydroxy-5-methoxy-8-(γ,γ-dimethylallyl)-flavanone (12), (2S)-7,4′-dihydroxy-5-methoxy-8-(γ,γ-dimethylallyl)-flavanone (13) and umbelliferone (14) from A. F. Immaturus were obtained. Three compounds (compounds 1113) from A. F. Immaturus were isolated for the first time from this medicinal plant. Among isolated compounds, ten compounds (compounds 1, 2, 612, 13) showed an inhibitory effect on the proliferation and the capillary-like tube formation of TR-LE cells. In addition, all compounds except compound 12 showed selective inhibition of the proliferation of TR-LE cells compared to Hela and Lewis lung carcinoma (LLC) cells. These compounds might offer clinical benefits as lymphangiogenesis inhibitors and may be good candidates for novel anti-cancer and anti-metastatic agents.
  • Young Hoon Sul, Myung Sun Lee, Eun Young Cha, Phuong Thien Thuong, Ngu ...
    2013 年 36 巻 1 号 p. 158-164
    発行日: 2013/01/01
    公開日: 2013/01/08
    ジャーナル フリー
    Hepatocellular carcinoma (HCC) is the most common type of liver cancer with high mortality worldwide. Traditional chemotherapy for HCC is not widely accepted by clinical practitioners because of its toxic side effects. Thus, there is a need to identify chemotherapeutic drugs against HCC. AMP-activated protein kinase (AMPK) is a biologic sensor for cellular energy status that acts a tumor suppressor and a potential cancer therapeutic target. The traditional Vietnamese medicinal plant Croton tonkinensis shows cytotoxicity in various cancer cells; however, its anticancer mechanism remains unclear. In this study, we determined whether the ent-kaurane diterpenoid ent-18-acetoxy-7β-hydroxy kaur-15-oxo-16-ene (CrT1) isolated from this plant plays a role as a chemotherapeutic drug targeting AMPK. CrT1 blocked proliferation in dose- and time-dependent manners in human hepatocellular carcinoma SK-HEP1 cells. CrT1 induced sub-G1 arrest and caspase-dependent apoptosis. CrT1 activated caspase-3, -7, -8, -9, and poly(ADP-ribose) polymerase, and its effect was inhibited by z-VAD-fmk suppressing caspase-3 cleavage. CrT1 induced increases in p53 and Bax levels but decreased Bcl2 levels. In addition, CrT1 resulted in increased translocation of cytochrome c into the cytoplasm. We showed that CrT1-activated AMPK activation was followed by modulating the mammalian target of rapamycin/p70S6K pathway and was inactivated by treating cells with compound C. Treatment with CrT1 and aminoimidazole carboxamide ribonucleotide (AICAR) synergistically activated AMPK. CrT1-induced AMPK activation regulated cell viability and apoptosis. These results suggest that CrT1 is a novel AMPK activator and that AMPK activation in SK-HEP1 cells is responsible for CrT1-induced anticancer activity including apoptosis.
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