Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
27 巻 , 4 号
選択された号の論文の33件中1~33を表示しています
Review
  • Toru Matsukawa, Kunizo Arai, Yoshiki Koriyama, Zhongwu Liu, Satoru Kat ...
    2004 年 27 巻 4 号 p. 445-451
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Since Sperry's work in the 1950s, it has been known that the central nervous system (CNS) neurons of lower vertebrates such as fish and amphibians can regenerate after axotomy, whereas the CNS neurons of mammals become apoptotic after axotomy. The goldfish optic nerve (ON) is one of the most studied animal models of CNS regeneration. Morphological changes in the goldfish retina and tectum after ON transection were first researched in the 1970s—1980s. Many biochemical studies of neurite outgrowth-promoting substances were then carried out in the 1980s—1990s. Many factors have been reported to be active substances that show increased levels during fish ON regeneration, as shown by using various protein chemistry techniques. However, there are very few molecular cloning techniques for studying ON regeneration after injury. In this review article, we summarize the neurite outgrowth-promoting factors reported by other researchers and describe our strategies for searching for ON regenerating molecules using a differential hybridization technique in the goldfish visual system. The process of goldfish ON regeneration after injury is very long. It takes about half a year from the start of axonal regrowth to complete restoration of vision. The process has been classified into three stages: early, middle and late. We screened for genes with increased expression during regeneration using axotomized goldfish retinal and tectal cDNA libraries and obtained stage-specific cDNA clones that were upregulated in the retina and tectum. We further discuss functional roles of these molecules in the regeneration processes of goldfish ON.
Current Topics
  • Hiroyuki Kamiya
    2004 年 27 巻 4 号 p. 452
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
  • Makoto Koizumi
    2004 年 27 巻 4 号 p. 453-456
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Novel 2′-O,4′-C-ethylene nucleosides have been synthesized as building blocks for antisense and antigene oligonucleotides. 2′-O,4′-C-Ethylene-bridged nucleic acids (ENATM) comprising 2′-O,4′-C-ethylene nucleosides have considerable affinity to complementary RNA and double-stranded DNA. Incorporation of 2′-O,4′-C-ethylene nucleosides into oligonucleotides dramatically increase their resistance against exonucleases. In this review, the properties of ENA oligonucleotides and some of their applications as antisense and antigene oligonucleotides are described.
  • Yasuo Komatsu
    2004 年 27 巻 4 号 p. 457-462
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Both hammerhead and hairpin ribozymes, which are well known as small catalytic RNAs, can catalyze either cleavage or ligation of RNA with sequence specificity. Although they have been applied to gene therapy and gene discovery, by cutting the mRNAs of target genes, these ribozymes normally do not have the ability to regulate their catalytic activities. Therefore, allosteric control has been applied to the ribozymes. Watson–Crick base pairing has superior ability in molecular recognition, and the sequence of nucleic acids makes diversity of molecules possible. Therefore, we have tried to construct a new ribozyme of which activity is regulated by a specific oligonucleotide. The allosteric ribozyme has potentials of not only regulation of activity but also a sensor for gene expression.
  • Fumi Nagatsugi, Shigeki Sasaki
    2004 年 27 巻 4 号 p. 463-467
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    The development of methods for targeted mutagenesis shows promise as an alternative form of gene therapy. Triple helix-forming oligonucleotides (TFOs) provide an attractive strategy for inducing mutations. Especially, alkylation of nucleobases with functionalized TFOs would have potential for site-directed mutation. Several studies have demonstrated that treatment of mammalian cells with TFOs can be exploited to introduce desired sequence changes and point mutations. This review summarizes targeted mutagenesis using reactive TFOs, including studies with photo reactive psolaren derivatives as well as a new reactive derivative recently developed by our group.
  • Musti Sree Rama Chandra Murty, Hiroshi Sugiyama
    2004 年 27 巻 4 号 p. 468-474
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Chemical substances that can recognize and bind DNA in a sequence-specific manner have enormous importance in modern biology and medicine. When covalently linked, hairpin polyamides made up of N-methylpyrrole (Py) and N-methylimidazole (Im) can bind to DNA in a sequence-specific manner. An Im opposite a Py recognizes and binds G:C from C:G, whereas a Py opposite an Im recognizes and binds to C:G. A Py–Py pair degenerately binds to T:A or A:T, whereas a hydroxypyrrole opposite a Py recognizes and binds to T:A from A:T, and vice versa. A variant in this recognition is the β-alanine (β-ala–β-ala) pair, which also degenerately binds to A:T or T:A. The hairpin polyamides are cell permeable and bind to DNA at nanomolar concentrations, with binding coefficients similar to those of transcription factors. This review comprehensively discusses the current literature on using the sequence-specific recognition ability of the polyamides to study various DNA–protein interactions.
  • Hiroyuki Kamiya
    2004 年 27 巻 4 号 p. 475-479
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Oligodeoxyribonucleotides containing 8-hydroxyguanine and 2-hydroxyadenine, purine lesions produced in cells by reactive oxygen species, were synthesized and inserted into vector DNAs to introduce each lesion at a predetermined site. The manipulated DNAs were transfected into living cells, and the mutants induced by each DNA lesion were collected and analyzed. In addition, the mutations induced by damaged DNA precursors with the two oxidized purine bases were studied by the use of chemically synthesized nucleoside triphosphates. In this review article, the author summarizes the mutagenic potentials of the two oxidized purine bases, by focusing on experiments examined by the author and his collaborators.
  • Hiroshi Ide, Mitsuharu Kotera
    2004 年 27 巻 4 号 p. 480-485
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Reactive oxygen species from endogenous and environmental sources induce oxidative damage to DNA, and hence pose an enormous threat to the genetic integrity of cells. Such oxidative DNA damage is restored by the base excision repair (BER) pathway that is conserved from bacteria to humans and is initiated by DNA glycosylases, which simply remove the aberrant base from the DNA backbone by hydrolyzing the N-glycosidic bond (monofunctional DNA glycosylase), or further catalyze the incision of a resulting abasic site (bifunctional DNA glycosylase). In human cells, oxidative pyrimidine lesions are generally removed by hNTH1, hNEIL1, or hNEIL2, whereas oxidative purine lesions are removed by hOGG1. hSMUG1 excises a subset of oxidative base damage that is poorly recognized by the above enzymes. Unlike these enzymes, hMYH removes intact A misincorporated opposite template 8-oxoguanine during DNA replication. Although hNTH1, hOGG1, and hMYH account for major cellular glycosylase activity for inherent substrate lesions, mouse models deficient in the enzymes exhibit no overt phenotypes such as the development of cancer, implying backup mechanisms. Contrary to the mouse model, hMYH mutations have been shown to lead to a multiple colorectal adenoma syndrome and high colorectal cancer risk. For cleavage of the N-glycosidic bond, bifunctional DNA glycosylases (hNTH1, hNEIL1, hNEIL2, and hOGG1) use Lys or Pro for direct attack on sugar C1′, whereas monofunctional DNA glycosylases (hSMUG1 and hMYH) use an activated water molecule. DNA glycosylases for oxidative damage, if not all, are covalently trapped by DNA containing 2-deoxyribonolactone or oxanine. Thus, the depletion of functional DNA glycosylases using covalent trapping may reduce the BER capacity of cancer cells, hence potentiating the efficacy of anticancer drugs or radiation therapy.
Analytical Biochemistry
Regular Articles
  • Yasushi Hori, Manami Fujisawa, Kenji Shimada, Akira Oda, Shinichiro Ka ...
    2004 年 27 巻 4 号 p. 486-491
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    We have established a new method of HPLC analysis for the rapid separation from human serum and the quantification of 4-O-methylpyridoxine (MPN), which is contained in Ginkgo biloba seeds, and which, when consumed in large amounts, causes vomiting and convulsions. As a result of using IPCC-MS3 (GL Science, Tokyo, Japan), an ion-pair reagent, in the mobile phase, we succeeded in separating MPN in the deproteinized serum sample which was introduced directly onto the reverse-phase HPLC column. For the calibration curve of MPN standard solution, prepared with fluorescence detection at an excitation wavelength of 290 nm and an emission wavelength of 400 nm, a good linear relationship was obtained within the HPLC injection range of 10 ng—10 pg (in terms of the injected sample concentration, range: 1.0 μg/ml—1 ng/ml), allowing the detection of minute amounts, with the limit of detection (concentration of injected sample: 500 pg/ml) being 5 pg. In addition, when MPN solution was added to human reference serum to give a concentration of 0.002 μg/ml, the mean recovery rate was 92.5%, with RSD=7.09% (n=5). The time required for one analysis using this method is approximately 30 min, and thus it offers the advantages of greater speed and superior analytical sensitivity over the conventional methods, which require solid-phase extraction. We employed our new method to determine both the serum levels of MPN in 5 patients with Ginkgo biloba seed poisoning and the levels of free-form MPN in such seeds obtained in 8 regions of Japan.
Biochemistry/Molecular Biology
Regular Articles
  • Fusako Nagai, Eriko Kato, Hiro-omi Tamura
    2004 年 27 巻 4 号 p. 492-495
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    To explore the effects of oxidative stress on metabolizing enzymes in blood cells, we examined the effect of hydrogen peroxide (H2O2) on glutathione S-transferase (GST) and cytochrome P450 (CYP) expression in a human erythroleukemic cell line, K562. After adding H2O2 (up to 100 μM) to the culture medium of K562 cells, the expression levels of GST and CYP enzymes were monitored by RT-PCR. The expression of GSTP1 and CYP3A4 was induced by oxidative stress. Quantitative PCR and immunoblot analysis revealed a 3- to 4-fold increase in GSTP1 and CYP3A4 mRNA, and a 2- to 3-fold increase in GSTP1 and CYP3A4 protein levels, 3 d after the addition of 100 μM H2O2. Induction was H2O2 dose-dependent and also depended on the length of culture. Our results suggest that oxidative stress may affect GST and/or CYP expression in human blood cells, which may alter the metabolism of drugs and xenobiotics and thus, the toxicity of these compounds to the blood cells.
  • Midori Suenaga, Masamichi Kuwajima, Toshiki Himeda, Kayoko Morokami, T ...
    2004 年 27 巻 4 号 p. 496-503
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Juvenile visceral steatosis (JVS) mice, novel animal models of systemic carnitine deficiency, exhibit a remarkably increased number of mitochondria in their cardiac myocytes. To date, however, there has been no reported investigation of the molecular mechanism of this increased number of mitochondria. Here, we analyzed the gene expression profile from the hearts of JVS and control mice by Affymetrix GeneChip analysis representing 34323 genes. We found that 176 genes, containing 93 known genes and 83 novel genes, were up-regulated in JVS mice compared with control mice, and 167 genes, containing 67 known genes and 100 novel genes, were down-regulated in JVS mice compared with control mice. We found several interesting molecular aspects that have not yet been identified in the hearts of JVS mice, including down-regulation of a number of ion channels and up-regulation of regulators involved in cell cycle progression. This genome-wide analysis should contribute to a greater understanding of the molecular mechanism of mitochondrial biogenesis in the heart of JVS mouse and provide a strategy for identifying novel genes involved not only in mitochondrial biogenesis but also in cardiac hypertrophy.
Notes
Pharmacology
Regular Articles
  • Je-Tae Woo, Hiroshi Nakagawa, Michitaka Notoya, Takayuki Yonezawa, Nob ...
    2004 年 27 巻 4 号 p. 504-509
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Although quercetin has suppressed bone resorption in several animal studies, its target cells and the mechanism of its action related to bone resorption has not been fully elucidated. We investigated the effect of quercetin on the differentiation and activation of osteoclasts. We used cocultures of mouse spleen cells and ST2 cells, and cultures of osteoclast progenitor cells {M-CSF-dependent (MD) cells from mouse bone marrow and murine monocytic RAW 264 (RAW) cells}. Quercetin dose-dependently inhibited osteoclast-like (OCL) cell formation at 2—5 μM concentration in both the coculture and MD cell culture. Quercetin inhibited the increase of tartrate-resistant acid phosphatase (TRAP) activity of mononuclear preosteoclasts (pOCs) induced by receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL) in both MD and RAW cell cultures. Quercetin reversely induced the disruption of actin rings in OCLs. Quercetin also suppressed both pit formation induced by osteoclasts on dentine slices and PTH-stimulated 45Ca release in mouse long bone cultures. These results suggest that osteoclast progenitors as well as mature osteoclasts, are quercetin's target cells in relation to bone resorption, and that quercetin's suppressive effect on bone resorption results from both its inhibitory effect on the differentiation of osteoclast progenitor cells into pOCs and from its disruptive effect on actin rings in mature osteoclasts.
  • Kazuhisa Sugimoto, Takahisa Nishimura, Koji Nomura, Kenji Sugimoto, Ta ...
    2004 年 27 巻 4 号 p. 510-514
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    We studied the inhibitory effects of 4-hydroxyphenyl α-glucopyranoside (α-arbutin) on melanogenesis in cultured human melanoma cells, HMV-II, and in a three-dimensional cultured human skin model. α-Arbutin showed no inhibitory effect on HMV-II cell growth at a concentration below 1.0 mM. Melanin synthesis in cells treated with α-arbutin at 0.5 mM decreased to 76% of that in non-treated cells. The cellular tyrosinase activity of HMV-II cells also significantly decreased, while the expression of its mRNA was not affected. Melanin synthesis in a human skin model was also evaluated by the macro- and microscopic observation of its pigmentation as well as by quantitative measurements of melanin. Treatment of the human skin model with 250 μg of α-arbutin did not inhibit cell viability, while melanin synthesis was reduced to 40% of that in the control. These results indicate that α-arbutin is an effective and safe ingredient for skin-lightening.
  • Byung-Soo Koo, Seung-Il Lee, Jeoung-Hee Ha, Dong-Ung Lee
    2004 年 27 巻 4 号 p. 515-519
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    The present study was performed to evaluate the central nervous system inhibitory effects of the essential oil from SuHeXiang Wan (Storax pill), a prescription usually used for treating epilepsy in traditional Chinese medicine, on fragrance inhalation (aroma therapy). Preinhalation of the fragrance oil markedly delayed the appearance of pentylenetetrazole-induced convulsion, but showed weak activities on picrotoxin- and strychnine-induced convulsions, which implies this drug may inhibit the convulsion by GABAergic neuromodulation. This essential oil inhibited the binding of [3H]Ro15-1788, a selective antagonist for the benzodiazepine receptor and also the binding of [3H]flunitrazepam, a selective agonist for the receptor, in the presence of γ-aminobutyric acid (GABA) and NaCl, showing a positive GABA shift, which suggested the strong possibility of the agonistic activity of the essential oil to the GABA/benzodiazepine receptor complex in rat cerebral cortices. Furthermore, inhalation inhibited the activity of GABA transaminase as the inhalation period was lengthened. The GABA level was significantly increased and glutamate content was significantly decreased in mouse brain by preinhalation of the essential oil. The above results suggest that the anticonvulsive effect of this essential oil can also originate from the enhancement of GABA level in the mouse brain, because convulsion depends partially on GABA concentration which can be properly preserved by inhibiting GABA transaminase. Fragrance inhalation progressively prolonged the pentobarbital-induced sleeping time as inhalation time was lengthened and inhibited brain lipid peroxidation, to which the anticonvulsive action is attributed; this also supported the above results, confirming the inhibitory effects of the essential oil of SuHeXiang Wan on the CNS via the GABAergic system.
  • Shinji Miura, Kohei Yamada, Fumitaka Kano, Yuichi Yoshimura
    2004 年 27 巻 4 号 p. 520-523
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Background: To find a nucleoside with anti-angiogenic activity, we tried to screen an active compound from our nucleoside library. Materials and Methods: The compound inhibiting the growth of human umbilical vein endothelial cell (HUVEC) induced by the conditioned medium of lung carcinoma cell line PC-9 was screened. The antitumor activity of the compound was evaluated against murine sarcoma S-180 implanted onto chick embryo chorioallantoic membrane (CAM). Results: 9-(4-Thio-β-D-ribo-pentofuranosyl)guanine (4′-thioguanosine) was found to be a potent inhibitor of the growth of HUVEC. The growth of S-180 implanted onto CAM was also inhibited by 4′-thioguanosine whereas the in vitro growth of S-180 was not inhibited. The administration of 4′-thioguanosine in mice caused unexpected side effect which suggested neurotoxicity. Conclusions: Antitumor effect of 4′-thioguanosine on S-180 was suggested to be due to inhibition of tumor angiogenesis. Because of toxicity of 4′-thioguanosine in mice, further development of the derivatives which have lower toxicity is required.
  • Tsutomu Abe, Norio Takagi, Midori Nakano, Kouichi Tanonaka, Satoshi Ta ...
    2004 年 27 巻 4 号 p. 524-527
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    A possible involvement of inhibitory effects of monobromobimane (MBM), a thiol reagent, on the swelling and the release of cytochrome c in the isolated brain mitochondria was examined. MBM dose-dependently inhibited the calcium and phenylarsineoxide-induced mitochondrial swelling and cytochrome c release. Significant relationships between mitochondrial swelling and cytochrome c release were detected. Furthermore, effects of in vivo treatment with MBM on neuronal cell damage after transient (15 min) global ischemia in rats were examined. Infusion of MBM (1 or 3 μg/animal) to cerebral ventricles attenuated an increased number of TUNEL-positive cells and neuronal cell death in the hippocampal CA1 region at 72 h of reperfusion. These results suggest that MBM may have an ability to inhibit mitochondria-associated apoptotic pathways through attenuation of the mitochondrial swelling and the release of cytochrome c.
Notes
Toxicology
Regular Articles
  • Pottekkad Vijayan, Preethi Vijayaraj, Prashanth Haranahalli Chandrappa ...
    2004 年 27 巻 4 号 p. 528-530
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    The total alkaloid fractions of the methanolic extracts of the leaves, ripe fruits, roots, seeds and stem of Solanum pseudocapsicum were subjected to in-vitro cytotoxicity, short-term toxicity and long-term survival studies. All the five fractions exhibited potent activity. The total alkaloid fraction of leaves was found to be the most potent. The HT-29 cell line was the most sensitive to the fractions. The cytotoxic concentration (CTC50) values for all these fractions ranged between 0.39—0.91, 0.68—2.8, 0.92—3.56, 4.05—8.2, 3.28—5.65 and 0.95—5.55 μg/ml, respectively for HT-29, RD-228, A-549, HEp-2, B16F10 and Vero cell lines. In short-term toxicity studies, the fractions showed 50% viability at 93—128 μg/ml for DLA cells and 141—189 μg/ml for human lymphocytes. In the long-term survival studies on the cell lines RD-228, HEp-2 and Vero, cells retained their regenerative capacities at concentrations below 8 μg/ml. The total alkaloids of the plant, especially from the leaves merit further investigations to identify the active constituents in animal models.
Notes
Medicinal Chemistry
Regular Articles
  • Shigeki Sasaki, Fumie Kurosaki, Terushi Haradahira, Fumihiko Yamamoto, ...
    2004 年 27 巻 4 号 p. 531-537
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Two new 11C-labelled ligands, N-(3-(4-hydroxyphenyl)propyl)-3-(4-methoxyphenyl)propylamine ([11C]2) and N-(3-(4-hydroxyphenyl)butyl)-3-(4-methoxyphenyl)butylamine ([11C]3) were designed based on bis(phenylalkyl)amines (1) which have been reported as polyamine site antagonists with high-selectivity for NR1A/2B NMDA receptors, and radiolabelling of the corresponding phenol precursors with [11C]methyl iodide was readily accomplished. The in vitro inhibition experiments using rat brain slices showed that [11C]2 and [11C]3 share the binding sites with spermine and/or ifenprodil but not with CP-101,606, a highly potent NR2B-selective NMDA antagonist, and that divalent cations such as Zn2+ produced significant inhibition of both [11C]2 and [11C]3 bindings. Intravenous injection of [11C]3 in mice showed almost homogenous distribution throughout the brain. Attempts to block the tracer uptake of [11C]3 by pre-injection with the unlabelled 3 or spermine in rats were unsuccessful, but a small decrease in the cerebral uptake of [11C]3 by co-treatment with the unlabelled 3 was observed in a monkey PET study. The present findings indicate that none of these 11C-labelled analogues have potential for PET study of binding sites on the N-methly-D-aspartate (NMDA) receptors.
Pharmacognosy
Regular Articles
  • Kyung-Min Shin, Yang-Hee Kim, Wan-Su Park, Insug Kang, Joohun Ha, Jong ...
    2004 年 27 巻 4 号 p. 538-543
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    In an attempt to search for bioactive natural products exerting antiinflammatory activity, we have evaluated the effects of the methanol extract of the fruits of Kochia scoparia (L.) CHARD. (Chenopodiaceae) on lipopolysaccharide (LPS)-induced nitric oxide (NO), prostaglandin E2 (PGE2), and tumor necrosis factor (TNF)-α release by the macrophage cell line RAW 264.7. Our data indicate that this extract is a potent inhibitor of NO production and it also significantly decreased PGE2 and TNF-α release. Consistent with these observations, the protein and mRNA expression level of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 was inhibited by MeOH extracts of Kochia scoparia (KSM) in a dose-dependent manner. Furthermore, KSM inhibited the LPS-induced DNA binding activity of nuclear factor-κB (NF-κB), which was associated with prevention of the inhibitor κB degradation. These results suggest that the methanol extract of K. scoparia inhibits LPS-induced iNOS and COX-2 expression by blocking NF-κB activation.
  • Mi-Jeong Ahn, Kee-Dong Yoon, So-Young Min, Ji Suk Lee, Jeong Ha Kim, T ...
    2004 年 27 巻 4 号 p. 544-547
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    The bioassay-directed isolation of a marine brown alga, Ecklonia cava, afforded four phlorotannin derivatives, eckol (1), 8,8′-bieckol (2), 8,4′′′-dieckol (3), and phlorofucofuroeckol A (4). Among these compounds, 2 and 3 exhibited an inhibitory effect on human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) and protease. Specifically, they inhibited the RT more potently than the protease. The inhibitory activity of compound 2 (IC50, 0.51 μM) against HIV-1 RT was comparable to that of nevirapine (IC50, 0.28 μM), a reference compound. An enzyme kinetic assay showed that this compound inhibited the RNA-dependent DNA synthesis activity of HIV-1 RT noncompetitively against dUTP/dTTP with a Ki value of 0.78 μM. With respect to the homopolymeric template/primer, (rA)n(dT)15, 8,8′-bieckol (2) displayed an uncompetitive type of inhibition (Ki, 0.23 μM).
  • Eun-Mi Ahn, Norio Nakamura, Teruaki Akao, Tsutomu Nishihara, Masao Hat ...
    2004 年 27 巻 4 号 p. 548-553
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    In the course of our search for natural estrogenic compounds from medicinal plants, we found that the methanolic extract from the roots of Moghania philippinensis (Fabaceae) showed significant effects on the proliferation of MCF-7 cells (human breast cancer) and induction of β-galactosidase activity in a yeast two-hybrid assay. Through estrogenic activity-guided fractionation, we isolated several active flavonoids including prenylated ones. The CHCl3 fraction and its new constituent, 8-(1,1-dimethylallyl)genistein (9), appreciably increased the uterine weight in ovariectomized rats when administered orally for 14 consecutive days, in which compound 9 showed stronger estrogenic activity than genistein. Antiestrogenic activities were also examined based on the inhibition of MCF-7 cell proliferation and β-galactosidase activity in the yeast two-hybrid assay, mediated by 17β-estradiol. 5,7,3′,4′-Tetrahydroxy-6,8-diprenylisoflavone (6) showed the strongest antiestrogenic activity.
  • Toshiaki Makino, Yoshiaki Ito, Shin-ya Sasaki, Yuu Fujimura, Yoshihiro ...
    2004 年 27 巻 4 号 p. 554-558
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Gyokuheifu-san (GHS; Jade Windscreen Powder in English, Yupingfeng-san in Chinese) is an herbal formula in traditional Chinese medicine that consolidates the superficial resistance to protect from invasion by external pathogenic influences. We evaluated the preventive and curative effects of GHS on allergic rhinitis induced by Japanese cedar pollens in guinea pigs, since the pollen can be considered one of external pathogens indicated by GHS. Guinea pigs were sensitized by intranasal instillation of cedar pollen extract with alum twice a day for 7 d, and the animals were then forced to inhale the pollens for challenge once a week for 5 weeks. We administered GHS once a day for 2 weeks in the period of sensitization to evaluate its preventive effect, or for 2 weeks from the 2nd to the 4th week of pollen inhalation, once pollinosis had begun, to evaluate its curative effect on allergic rhinitis. GHS significantly suppressed the frequency of sneezing induced by pollens and tended to reduce nose-scratching behavior after ceasing its administration in both designs of medicinal treatment. Tranilast, which is an anti-allergic drug we used as a positive control, could not suppress these rhinitic symptoms. GHS appears to have non-symptomatic and non-allopathic effects on allergic rhinitis. Our results suggest that traditional medicines have their own characteristics different from modern medicines, and the original pharmacological experiments are important to evaluate traditional medicines scientifically.
Notes
  • Masaru Ogasawara, Hideyo Suzuki
    2004 年 27 巻 4 号 p. 578-582
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Tumor cell motility plays a crucial role in the establishment of tumor metastasis and is affected by a variety of host-derived factors during the event. Hepatocyte growth factor (HGF) is one of these factors and stimulates tumor cell migration remarkably. We previously reported that evodiamine has a marked inhibitory activity on tumor cell invasion and migration in vitro. In this study, the effects of evodiamine on HGF-induced invasion and migration of tumor cell lines, colon 26-L5 carcinoma, B16-F10 melanoma and Lewis lung carcinoma (LLC) were examined. HGF promoted invasive activity of tumor cell lines with maximal induction of 1.8 times at 30 ng/ml for colon 26-L5 and LLC cells, and 2.0 times at 10 ng/ml for B16-F10 cells. Evodiamine inhibited the HGF-stimulated tumor cell invasion and migration in a concentration-dependent manner, and achieved complete suppression at 30 μM in all of the cell lines tested. When tumor cells were seeded on fibronectin-coated plates with evodiamine, their spreading on the plate was obviously inhibited, while their adhesiveness to fibronectin was unaffected. Evodiamine showed a marginal effect on tumor cell growth in a 24-h incubation, although it exhibited a marked inhibition in an over 48-h incubation. These results suggest that evodiamine suppressed HGF-stimulated invasion and migration of tumor cells partly through inhibition of cell spreading.
  • Jun Yin, Yasuhiro Tezuka, Kyoji Kouda, Quan Le Tran, Tatsuro Miyahara, ...
    2004 年 27 巻 4 号 p. 583-586
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    After 60 MeOH and water extracts of natural crude drugs were screened for their ability to stimulate osteoblast proliferation, four MeOH extracts (Cynomorium songaricum, Drynaria fortunei, Lycium chinense, Rehmannia glutinosa) and seven water extracts (Cornus officinalis, Dendrobium nobile, Dioscorea spongiosa, Drynaria fortunei, Eucommia ulmoides, Lycium chinensis, Viscum coloratum) showed that potent activities were evaluated for inhibition of osteoclast formation. The results indicated that the water extract of D. spongiosa not only showed the strongest stimulation of osteoblast proliferation but also possessed potent inhibitory activity aganist osteoclast formation, whereas it showed lower cytotoxicity in osteoblast and bone marrow cells. A further in vivo experiment determined the antiosteoporotic activity of this extract, in which it inhibited the decrease in cancellous bone mineral content, cancellous bone mineral density, and cortical bone mineral content of the proximal tibia in ovariectomized rats.
Biopharmacy
Regular Articles
  • Kazuhiro Watanabe, Toshiya Jinriki, Juichi Sato
    2004 年 27 巻 4 号 p. 559-563
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Little is known about the regulatory effects of steroid hormones on the intestinal H+/oligopeptide transporter PEPT1. In the present study, we investigated the effects of progesterone and its related compounds on the uptake of cephalexin, a typical PEPT1 substrate, using the human intestinal cell line Caco-2. Caco-2 cell monolayers were cultured on plastic cell culture plates coated with rat tail collagen type I. The determination of cephalexin uptake was performed with HPLC. To investigate the effect of progesterone treatment on cephalexin uptake, the monolayers were incubated with cephalexin after progesterone treatment. Progesterone decreased cephalexin uptake in a concentration-dependent manner, and the IC50 value was calculated to be 2.3 μM. A significant decrease in cephalexin uptake was observed after progesterone treatment for 12 h, and the decrease was maximal when the monolayers were treated for 24—72 h. Cephalexin uptake was significantly inhibited by treatments with 17α-hydroxyprogesterone, norethisterone, and chlormadinone. By contrast, treatment with dehydroepiandrosterone, pregnenolone, estradiol, testosterone, hydrocortisone, and dexamethasone did not affect cephalexin uptake. The effects of progesterone and norethisterone treatment on the kinetic parameters of cephalexin uptake were investigated. The saturable component of cephalexin uptake was markedly inhibited by progesterone and norethisterone treatments. The Jmax of cephalexin uptake with progesterone and norethisterone treatments was significantly decreased compared with the control, whereas Km and Kd values were not affected. Therefore the quantitative decrease in PEPT1 density is considered to be one cause of the decrease in cephalexin uptake with progesterone and norethisterone treatments.
Notes
  • Takanori Kobayashi, Souichi Ikeno, Nobuo Hosokawa, Yoshimasa Uehara, M ...
    2004 年 27 巻 4 号 p. 587-590
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Destruxin E (DE), a cyclodepsipeptide isolated from fermentation broths of Metarhizium sp. MA324, inhibited the growth of v-Ki-ras-expressed pMAM-ras-REF (rasREF) cells in the suspension (anchorage-independent) culture (a) more strongly than that in the substratum-attached (anchorage-dependent) culture (b) or that of v-Ki-ras-unexpressed pMAM-ras-REF (REF) cells in the substratum-attached culture (c); the IC50 values of DE were 0.07 μM (a), 0.4 μM (b), and 1.2 μM (c). DE arrested G1 phase cell cycle progression of rasREF cells in the substratum-attached culture (b). In rasREF cells treated with DE for 72 h in suspension culture (a), the levels of cyclin D1, cyclin A, p27Kip1, and hyperphosphorylated Rb were decreased, but the levels of cdk4, cdk6, cdk2, p16INK4a, and p21Cip1 were not affected. Among these effects, the decrease in cyclin D1 was prominent. DE decreased the level of cyclin D1 in rasREF cells in the suspension culture (a) at 0.1 μM and in the substratum-attached culture (b) at 1 μM, while the level of cyclin D1 in REF cells in the substratum-attached culture (c) was not decreased at 1 μM. The extent of growth inhibition correlated with the decrease in cyclin D1. The level of cyclin D1 mRNA of rasREF cells in the suspension culture (a) was also decreased by DE. DE decreased cyclin D1 mRNA, resulting in inhibition of anchorage-independent growth of rasREF cells.
  • Yasutami Shigeta, Hiromichi Imanaka, Hideya Ando, Atsuko Ryu, Naoto Ok ...
    2004 年 27 巻 4 号 p. 591-594
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Linoleic acid (LA) is known to have a whitening effect on hyperpigmented skin, and is encapsulated in liposomes for topical application because of its low solubility in aqueous solution, although the effect of liposomalization of LA on the whitening activity has not been evaluated. In the present study, we evaluated the effect of liposomalization on the whitening activity of LA by using LA in ethanol, hydrogel containing LA, and hydrogel containing liposomal LA towards the UV-stimulated hyperpigmented dorsal skin of brownish guinea pigs. The whitening effect was far greater for hydrogel containing liposomal LA (0.1% w/w as a final concentration of LA) than for free LA in ethanol or hydrogel containing LA. Next, the whitening effect of LA was examined with UV-stimulated hyperpigmented human upper arm skin by using a hydrogel containing liposomal LA (0.1% LA) and non-liposomal LA (3.0, 10.0% LA). Liposomal LA (0.1%) showed a whitening effect comparable to 10.0% non-liposomal LA and was far more effective than 3.0% non-liposomal LA. These results indicate that liposomal formulations are favorable for the transdermal application of LA.
  • Takahiro Mukai, Kunihiro Mera, Koyo Nishida, Mikiro Nakashima, Hitoshi ...
    2004 年 27 巻 4 号 p. 595-597
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    Animal models prepared by treatment with carbon tetrachloride (CCl4) have been used to examine drug disposition in hepatic disorder. However, previous studies demonstrated that systemic administration of CCl4 impaired not only hepatic but also renal function. We recently reported that application of CCl4 to the rat liver surface produced hepatic damage without impairing renal function. In the present study, we examined the pharmacokinetics of phenol red in our developed rat model. The rats treated with CCl4 by liver surface application exhibited decreases in the biliary clearance of phenol red in comparison with normal rats from 0.54±0.03 to 0.31±0.06 ml/min, suggesting hepatic damage. In these rats, the renal clearance of phenol red did not decrease (0.50±0.16 ml/min vs. 0.46±0.07 ml/min in normal rats). On the other hand, oral and intraperitoneal treatments with CCl4 reduced not only the biliary clearance of phenol red (0.34±0.03 ml/min in p.o. treated rats, 0.18±0.01 ml/min in i.p. treated rats) but also the renal clearance (0.26±0.05 ml/min in p.o. treated rats, 0.18±0.06 ml/min in i.p. treated rats) as compared with normal rats. These findings indicate that the rat model of liver damage prepared by liver surface application of CCl4 is useful to investigate the effects of hepatic disorder on the pharmacokinetics of drugs.
Errata
  • 2004 年 27 巻 4 号 p. 598
    発行日: 2004年
    公開日: 2004/04/01
    ジャーナル フリー
    The Editorial Committee of this Journal wishes to acknowledge with regret that “Proteomic Analysis of Proteins that Binds Specifically to the Homologous Repeat Regions of White Spot Syndrome Virus” by Qin Li, Feng Yang, Jinghai Zhang, and Yingjie Chen which was published in Vol. 26 (2003), p. 1517 of this Journal, has been found to contain no reproducible results, according to proposal by the authors.
    Thus it should be noted that the above-mentioned paper by Qin Li, Feng Yang, Jinghai Zhang, and Yingjie Chen should be totally deleted.
    For this reason, as of February 12, 2004, this article has been removed from this Journal.
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