Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
27 巻 , 10 号
選択された号の論文の43件中1~43を表示しています
Review
  • Hiroko Shibata, Shinsaku Nakagawa, Tadanori Mayumi, Yasuo Tsutsumi
    2004 年 27 巻 10 号 p. 1483-1488
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    With the success of human genome projects, the focus of life science research has shifted to the functional and structural analyses of proteins, such as disease proteomics. These structural and functional analyses of expressed proteins in the cells and/or tissues are expected to contribute to the identification of therapeutically applicable proteins for various diseases. Thus, pharmaco-proteomic based drug development for protein therapies is most noticed currently. However, there is a clinical difficulty to use almost bioactive proteins, because of their very low stability and pleiotropic actions in vivo. To promote pharmaco-proteomic based drug development for protein therapies to various diseases, we have attempted to establish a system for creating functional mutant proteins (muteins) with desired properties, and to develop a site-specific polymer-conjugation system for further improving the therapeutic potency of proteins. In this review, we are introducing our original protein-drug innovation systems mentioned above.
  • Sumio Ohtsuki
    2004 年 27 巻 10 号 p. 1489-1496
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    The blood–brain barrier (BBB) segregates the circulating blood from interstitial fluid in the brain, and restricts drug permeability into the brain. Our latest studies have revealed that the BBB transporters play important physiological roles in maintaining the brain milieu. The BBB supplies creatine to the brain for an energy-storing system, and creatine transporter localized at the brain capillary endothelial cells (BCECs) is involved in BBB creatine transport. The BBB is involved in the brain-to-blood efflux transport of the suppressive neurotransmitter, γ-aminobutyric acid, and GAT2/BGT-1 mediates this transport process. BCECs also express serotonin and norepinephrine transporters. Organic anion transporter 3 (OAT3) and ASCT2 are localized at the abluminal membrane of the BCECs. OAT3 is involved in the brain-to-blood efflux of a dopamine metabolite, a uremic toxin and thiopurine nucleobase analogs. ASCT2 plays a role in L-isomer-selective aspartic acid efflux transport at the BBB. Dehydroepiandrosterone sulfate and small neutral amino acids undergo brain-to-blood efflux transport mediated by organic anion transporting polypeptide 2 and ATA2, respectively. The BBB transporters are regulated by various factors, ATA2 by osmolarity, taurine transporter by TNF-α, and L-cystine/L-glutamic acid exchange transporter by oxidative stress. Clarifying the physiological roles of BBB transport systems should give us important information allowing the development of better CNS drugs and improving our understanding of the relationship between CNS disorders and BBB function.
Biochemistry/Molecular Biology
Regular Articles
  • Heun-Sik Lee, Mie-Hee Park, Suk-Jin Yang, Hai-Young Jung, Sang-Soon By ...
    2004 年 27 巻 10 号 p. 1497-1503
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Trichostatin A (TSA) and S-adenosyl-L-homocysteine (AdoHcy) have been reported to affect histone modifications. To investigate the effects of two drugs that can reportedly affect chromatin remodeling, we analyzed the gene expression profiles of TSA and AdoHcy in a gastric cancer cell line using 14 K cDNA microarray. The significant analysis of microarray (SAM) identified 98 and 43 differentially expressed genes in TSA and AdoHcy treated sets, respectively, and selected genes were functionally classified. In the gastric cancer cell line, genes related to cell communication, cell growth/maintenance, and morphogenesis were highly expressed with TSA, and genes with cell growth/maintenance, metabolism, oxidoreductase activity were upregulated with AdoHcy. Genes downregulated with TSA included those controlling the cell cycle, cell growth/proliferation, DNA binding, and metabolism, whereas genes involved in calcium signaling, cell growth/proliferation, and metabolism were downregulated with AdoHcy. Furthermore, we identified the genes commonly expressed in both drug treatments. Compared to TSA, AdoHcy did not induce apoptosis in the SNU-16 gastric cancer cell line, and RT-PCR was performed for selective genes to confirm the microarray data. This gene expression profile analysis with TSA and AdoHcy should contribute to a greater understanding of the molecular mechanism of chromatin remodeling and cancer, and provide candidate genes for further studies involving the roles of histone modifications in gastric cancer.
  • Sungwook Chae, Ju Sun Kim, Kyoung Ah Kang, Hyung Deuk Bu, Youngki Lee, ...
    2004 年 27 巻 10 号 p. 1504-1508
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    The antioxidant property of jionoside D, isolated from Clerodendron trichotomum (Verbenaceae), was investigated. This compound showed scavenging activity of intracellular reactive oxygen species and of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, as well as lipid peroxidation inhibitory activity. This radical scavenging activity of jionoside D protected the cell viability of Chinese hamster lung fibroblast (V79-4) cells exposed H2O2. Furthermore, jionoside D reduced the apoptotic cells induced by H2O2, as demonstrated by the decreased number of sub G1 hypo-diploid cells and apoptotic body formation. However, it increased the activities of cellular antioxidant enzymes, superoxide dismutase and catalase. Taken together, these findings suggest that jionoside D, isolated from C. trichotomum, exhibits antioxidant properties.
  • Naosuke Nakamura, Kenji Katano, Shinya Toba, Akira Kurosaka
    2004 年 27 巻 10 号 p. 1509-1514
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Polypeptide N-acetylgalactosaminyltransferases (GalNAc-transferases) catalyze the initial reaction of mucin-type O-glycosylation. Here, we report the first biochemical characterization of one of the Drosophila GalNAc-transferases, dGalNAc-T3. This enzyme retains conserved motifs essential for the catalytic activity, but is a novel isozyme in that it has several inserted sequences in its lectin-like domain. Northern hybridization analysis of this isozyme identified a 2.5-kb mRNA in Drosophila larva. Biochemical characterization was carried out using the recombinant soluble dGalNAc-T3 expressed in COS7 cells. dGalNAc-T3, which required Mn2+ for the activity, had a pH optimum ranging from pH 7.5 to 8.5, and glycosylated most effectively at 29—33 °C. Its Km for UDP-GalNAc was 10.7 μM, which is as low as that of mammalian isozymes. dGalNAc-T3 glycosylated the peptides containing a sequence of XTPXP or TTAAP most efficiently. The enzyme was irreversibly inhibited by p-chloromercuriphenylsulphonic acid, indicating the presence of essential Cys residues for the activity.
Notes
  • Yasuhiro Morita, Eiko Matsumura, Toshihiro Okabe, Toru Fukui, Tatsuhik ...
    2004 年 27 巻 10 号 p. 1666-1669
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    β-Dolabrin, γ-thujaplicin, and 4-acetyltropolone, the components of Aomori Hiba (Thujopsis dolabrata SIEB. et ZUCC. var. hondai MAKINO), showed antifungal activity on seven kinds of plant-pathogenic fungi, antibacterial activity against two kinds of Legionella sp., and in vitro cytotoxic effect on murine P388 lymphocytic leukemia cell line. Firstly, β-dolabrin, γ-thujaplicin and 4-acetyltropolone had clear antifungal activity against seven kinds of plant-pathogenic fungi tested. In particular, β-dolabrin and 4-acetyltropolone showed strong antifungal activity against Pythium aphanidermatum IFO 32440, with minimum inhibitory concentration (MIC) values of 6.0 μg/ml. Secondly, β-dolabrin, γ-thujaplicin and 4-acetyltropolone had obvious growth-inhibitory effect on two kinds of Legionella sp. 4-Acetyltropolone especially had strong antibacterial activity toward Legionella pneumophila SG 1, and its MIC value was 3.1 μg/ml. These three compounds showed cytotoxic effects against murine P388 lymphocytic leukemia cell line in vitro. The cytotoxic effect of three compounds in the murine P388 lymphocytic leukemia cell line were clear when cell growth was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. At 48 h after treatment, γ-thujaplicin and 4-acetyltropolone at 0.63 μg/ml inhibited cell growth of murine P388 lymphocytic leukemia by 85% and 65%, respectively. At the same time after treatment, the growth of the murine P388 lymphocytic leukemia cell line was completely suppressed by the three compounds at concentrations higher than 5.0 μg/ml. Among these three compounds, γ-thujaplicin had the strongest cytotoxic activity on the growth of this tumor cell line in vitro.
Pharmacology
Regular Articles
  • Wan-Liang Lu, Qiang Zhang, Li Zheng, Hua Wang, Rong-Yu Li, Li-Feng Zha ...
    2004 年 27 巻 10 号 p. 1515-1520
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Ketoprofen is a nonsteroidal anti-inflammatory drug (NSAID) orally effective in treating fever, pain, and inflammation but gastrointestinal side effects were observed. Preparation of ketoprofen β-cyclodextrin inclusion complexes was to increase the solubility and reduce the irritation. The complexes were prepared and preliminarily confirmed using X-ray diffraction and dissolution test. Antipyretic, analgesic and anti-inflammatory models were induced by 10% yeast using rabbits, 0.8% acetic acid using mice and 1% carrageenin using rats, respectively. Results showed that the dissolution rate of ketoprofen was significantly improved by complexation. X-Ray diffraction pattern of the complexes exhibited a diffuse pattern that differed from that of physical mixture of ketoprofen and β-cyclodextrin. Ketoprofen markedly inhibited the fever reactions at a single dose of 2 mg/kg as follows: 64.53% (inhibition rate %) at 1h for ketoprofen, 73.04% at 1 h for ketoprofen β-cyclodextrin inclusion complexes, respectively. Alleviating pain reaction rates following a single dose of 8 mg/kg at 20 min were 39.25% for the inclusion complexes and 26.72% for ketoprofen, respectively. Inhibition rates to rat edema following a single dose of 5 mg/kg at 1 h were 39.47% for the inclusion complexes and 23.86% for ketoprofen. Results for antipyretic, analgesic and anti-inflammatory activities showed that the rapid and stronger effects were found in the treatment group of ketoprofen β-cyclodextrin inclusion complexes in comparison with those of free ketoprofen.
  • Hye-Young Shin, Tae-Yong Shin, Sang-Wan Seo, Hyo-Jin An, Yong-Taek Kwo ...
    2004 年 27 巻 10 号 p. 1521-1526
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    WooKiEum (WKE) has been used for the purpose of the development of increased immune-system strength in Korea. In the present study, we examined the anti-immobility effect of WKE on the forced swimming test (FST), and then measured blood biochemical parameters related to fatigue: glucose (Glc), blood urea nitrogen (BUN), lactic dehydrogenase (LDH), creatinine, and total protein (TP). WKE (0.1, 1 g/kg) was administered orally to mice for 7 d. After 2 d, the immobility time was decreased in the WKE-administered group. After 7 d, the immobility time was significantly decreased in the WKE-administered group (64.6±9.0 s for 0.1 g/kg) in comparison with the control group (101.3±32.7 s). In addition, amount of Glc in the blood serum was increased, whereas the contents of BUN, LDH and TP decreased in the WKE-administered group. Next, we investigated the effect of WKE on the production of cytokines in a human T-cell line, MOLT-4 cells and mouse peritoneal macrophages. WKE (1 mg/ml) significantly increased interferon (IFN)-γ and TNF-α production compared with the media control (about 2.2-fold for IFN-γ, about 1.7-fold for TNF-α, p<0.05) after 24 h. WKE increased the protein expression of IFN-γ in MOLT-4 cells. These results suggest that WKE may be useful in immune function improvement.
  • Chun-Ling Zhang, Li-Jun Wu, Shin-ichi Tashiro, Satoshi Onodera, Takash ...
    2004 年 27 巻 10 号 p. 1527-1531
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Oridonin, an active component isolated from Rabdosia rubescences, has been reported to exhibit antitumor effects, but little is known about its molecular mechanisms of action. In this study, the growth-inhibitory activity of oridonin for L929 cells is in time- and dose-dependent manner. After treatment with various concentrations of oridonin for 12 h, the majority of L929 cells underwent apoptosis as measured by an LDH activity-based assay. Although apoptotic bodies were observed in oridonin-treated L929 cells, DNA fragmentation as a hallmark of apoptosis was not found. The pan-caspase inhibitor, z-VAD, and caspase-3 inhibitor, z-DEVD, sensitized L929 cells to oridonin, however, a PARP inhibitor (DPQ) effectively blocked oridonin-induced cell death. After 12 h treatment, PARP proenzyme was significantly cleaved. This result indicated that oridonin-induced L929 cell death required PARP degradation in a caspase-independent manner. In addition, an MEK/ERK inhibitor (PD98059) markedly blocked oridonin-induced cell death, whereas a p38 inhibitor (SB203580) and JNK inhibitor (SP600125) weakly protected the cells against death. Treatment with 41.2 μM oridonin for 12 h induced significant and persistent ERK activation and p38 inactivation in L929 cells without evident changes in the protein levels. The responsiveness of ERK and p38 to oridonin suggests the involvement of these kinases in this apoptotic process. Moreover, oridonin increased the ratio of Bax/Bcl-2 protein expression, whereas it had no effect on the expression of Bcl-xL. These results indicate that regulation of the Bcl-2 and MAPK families maybe the effector mechanisms of oridonin-induced L929 cell death, independent of the caspase pathway.
  • Li Gan, Xianliang Xin, Meiyu Geng, Jian Ding
    2004 年 27 巻 10 号 p. 1532-1536
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Two conjugates of marine polysaccharide (MPS) and bovine serum albumin (BSA) were prepared using two methods, periodate oxidation and reductive amination, with the intent of enhancing its immunogenicity. Sera samples from Balb/c mice immunized with the products named MPS–BSAp and MPS–BSAr respectively were evaluated by enzyme-linked-immunosorbent assay (ELISA). The results showed that mice immunized with MPS–BSAp produced antibodies not against MPS but rather against MPS–BSAp, while the mice immunized with MPS–BSAr produced high titer antibodies only specific for MPS. The difference was attributed to the fact that the epitopes of MPS had been changed in the coupling process by periodate oxidation. A mouse immunized with MPS–BSAr was chosen to prepare monoclonal antibodies (mAbs) specific for polysaccharide MPS. A hybridoma cell line that secreted monoclonal antibody recognizing specifically polysaccharide MPS was established.
  • Rong Lin, Juntian Liu, Weijie Gan, Guangde Yang
    2004 年 27 巻 10 号 p. 1537-1543
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Inflammation plays a pivotal role in the formation of atherosclerosis. In addition to being a risk marker for cardiovascular diseases, the role of C-reactive protein (CRP) in atherogenesis has been supported by more recent data. CD40–CD40L system is proven to be an important mediator of several auto-immune and chronic inflammation diseases. Interruption of CD40–CD40L signaling pathway not only reduces the initiation and progression of atherosclerotic lesions, but also modulates plaque architecture. By using a flow cytometry and western blotting, we found that incubation of human umbilical vein endothelial cells (HUVECs) with CRP resulted in a time- and dose-dependent increase in the cell-surface expression of CD40 and CD40L. In addition, CRP (25 μg/ml) increased gelatinolytic activities of MMP-2 and MMP-9. Anti-CD40 antibody significantly reversed the upregulated activities of MMP-2 and MMP-9 induced by CRP with gelatin zymography. Furthermore, lovastatin (10−7, 10−6, 10−5 mol/l) and fenofibrate (5×10−5, 10−4, 2×10−4 mol/l) significantly diminished the expression of CD40, CD40L and gelatinase activities (MMP-2, MMP-9) induced by CRP in HUVECs. In conclusion, our data provide evidence to support the direct pro-inflammatory effects of CRP via CD40–CD40L signaling pathway involved in the pathogenesis of atherosclerosis, and lovastatin and fenofibrate possess anti-inflammatory effects independent of their lipid-lowering action.
  • Kazuhiko Oishi, Tomokazu Yoshizumi, Noriyasu Seki, Masaatsu K. Uchida
    2004 年 27 巻 10 号 p. 1544-1548
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    The changes in the reorganization of actin filaments during desensitization of secretion were investigated by transfecting RBL-2H3 cells with cDNA encoding the human m3 muscarinic acetylcholine receptors (RBL-m3 cells). Incubation of RBL-m3 cells with 10—100 μM carbachol in Ca2+-free medium developed membrane ruffling. When the cells were desensitized under the condition where desensitization of carbachol-induced secretion occurred, desensitized cells failed to develop membrane ruffling with the subsequent addition of carbachol. These results suggest that m3 muscarinic receptor-mediated desensitization of secretion involves negative regulation of actin reorganization leading to membrane ruffling.
  • Toshikazu Tsuji, Toshinobu Kato, Masahiro Kimata, Toru Miura, Isao Ser ...
    2004 年 27 巻 10 号 p. 1549-1554
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    In the present study, we examined the inhibitory effects of the β2-adrenoceptor agonists isoproterenol, salbutamol, fenoterol, and clenbuterol, on the release of chemical mediators from cultured human mast cells after prolonged treatment with the agonists. Although preincubation of sensitized mast cells for 10 min with β2-adrenoceptor agonists potently inhibited mediator release, prolongation of the preincubation period up to 240 min attenuated the inhibition. The attenuation of histamine release inhibition was potent when compared with that of prostaglandin D2 (PGD2) and cysteinyl leukotriene (LT) release inhibition. In contrast, forskolin inhibited mediator release and the inhibition increased gradually in proportion to the preincubation period. The reduced inhibition by the β2-adrenoceptor agonists was compensated for by simultaneous treatment with cholera toxin. The β2-adrenoceptor agonists elevated intracellular cAMP levels after 10-min incubation and the elevated levels were almost comparable to those after 240-min incubation. Forskolin elevated the intracellular cAMP levels more potently after incubation for 240 min than after 10 min. When mast cells were incubated for 3 d with the β2-adrenoceptor agonists, similar attenuation of mediator release inhibition was observed. Elevation of intracellular cAMP levels was also attenuated, although β2-adrenoceptor mRNA expression was potentiated. The present results collectively indicate that the attenuation of mediator release inhibition by β2-adrenoceptor agonists under the present experimental conditions involves uncoupling between β2-adrenoceptors and Gs proteins. Furthermore, the β2-adrenoceptor desensitization causes differential attenuating effects on the inhibition of histamine, PGD2, and LT release, suggesting that downstream events involved in each inhibitory pathway have different sensitivity to receptor desensitization.
  • Yong Liu, Wei Li, Peng Li, Mai-Cun Deng, Sheng-Li Yang, Ling Yang
    2004 年 27 巻 10 号 p. 1555-1560
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    The intestinal bacterial metabolites of ginsenosides are responsible for the main pharmacological activities of ginseng. The purpose of this study was to find whether these metabolites influence hepatic metabolic enzymes and to predict the potential for ginseng–prescription drug interactions. Utilizing the probe reaction of CYP3A activity, testosterone 6β-hydroxylation, the effects of derivatives of 20(S)-protopanaxadiol and 20(S)-protopanaxatriol families on CYP3A activity in rat liver microsomes were assayed. Our results showed that ginsenosides from the 20(S)-protopanaxadiol and 20(S)-protopanaxatriol family including Rb1, Rb2, Rc, Compound-K, Re, and Rg1 had no inhibitory effect, whereas Rg2, 20(S)-panaxatriol and 20(S)-protopanaxatriol exhibited competitive inhibitory activity against CYP3A activity in these microsomes with the inhibition constants (Ki) of 86.4±0.8 μM, 1.7±0.1 μM, and 3.2±0.2 μM, respectively. This finding demonstrates that differences in their chemical structure might influence the effects of ginsenosides on CYP3A activity and that ginseng-derived products might have potential for significant ginseng–drug interactions.
  • Jungsook Cho, Hyeong-Kyu Lee
    2004 年 27 巻 10 号 p. 1561-1564
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    The present study evaluated the effect of wogonin, a flavonoid originated from the root of Scutellaria baicalensis GEORGI, on focal ischemic brain injury in rats. Focal brain ischemia was induced by the permanent occlusion of middle cerebral artery (pMCAO) for 24 h with a silicone rubber cylinder inserted through the right internal carotid artery. We found that wogonin, intraperitoneally administered at a dosage of 20 mg/kg at 30 min before and 4 h after the surgery, reduced the pMCAO-induced infarct areas in the cerebral cortex as well as in the striatum. The total volume of infarction was significantly reduced by the treatment with wogonin. In addition, wogonin was found to significantly improve the pMCAO-induced behavioral deficits at 24 h after the surgery. Taken together, these results demonstrate that wogonin inhibits ischemic brain injury and improves behavioral dysfunction caused by pMCAO. These findings, along with previous reports demonstrating the neuroprotective effects of wogonin, provide strong pharmacological basis for the use of wogonin or Scutellaria baicalensis in the treatment of stroke.
  • Shinjiro Kobayashi, Miho Suzuki, Hiroshi Tsuneki, Ryoji Nagai, Seikoh ...
    2004 年 27 巻 10 号 p. 1565-1571
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Action of Nε-(carboxymethyl)lysine (CML) adduct, an advanced glycation end product, was investigated on neovascularization of cultured choroidal explants in streptozotocin (STZ)-diabetic rat. The choroidal explants of early (4 weeks after an injection of 60 mg/kg STZ) and advanced (8 months after the STZ injection) diabetic rats, and age-matched normal rats were cultured in fibrin gel with Dulbecco's modified Eagle medium containing fetal bovine serum. The number of budded microvessel-like structures was counted and used as an index of in vitro neovascularization. Choroidal explants in the early diabetic stage released vascular endothelial growth factor (VEGF) and tended to increase tumor necrosis factor (TNF) α and platelet-derived growth factor (PDGF)-B, and concomitantly facilitated growth of sprout and buds, compared to the normal control. When choroidal explants were stimulated with CML-human serum albumin (HSA), its releasing effect was in the order VEGF>TNFα>PDGF-B. CML-HSA and CML-bovine serum albumin augmented the neovascularization in the cultured diabetic explant and their actions did not virtually differ. A monoclonal anti-CML antibody (6D12) inhibited the neovascularization in the advanced diabetes greater than that in the early diabetes. Inhibitory actions of anti-VEGF and anti-TNFα antibodies on the neovascularization were similar to that of the anti-CML antibody in the diabetes. In conclusion, CML adducts were accumulated and over-produced the actions of VEGF, TNFα and PDGF-B in the choroidal explant during diabetes in an age-dependent manner. TNFα and VEGF are likely to play a predominant role for the CML-induced choroidal neovascularization.
Notes
  • Eric Brian Hoie, Timothy Robert McGuire, Patricia Mary Leuschen, Terre ...
    2004 年 27 巻 10 号 p. 1670-1673
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Vascular endothelium is a major target for the inflammatory damage that occurs with multiple organ dysfunction associated with sepsis and other trauma. The growing appreciation of endothelium as a target of inflammation has obscured the importance of these cells as a source of inflammatory mediators. In the following study we evaluated the ability of tumor necrosis factor-alpha (TNF) to induce the synthesis of complement component C3 in human umbilical vein endothelial cells (HUVEC) and whether pentoxifylline (PTX) could reduce C3 expression. Confluent monolayers of HUVEC were treated with increasing concentrations of TNF with and without two concentrations of PTX. Concentrations of C3 were determined every 48 h for 144 h in cellular supernatants and C3 mRNA was amplified using RT-PCR. TNF increased C3 release from HUVEC in a concentration dependent manner. PTX added at the same time as TNF significantly reduced C3 release at the 96 h time point. Consistent with data on C3 release PTX inhibited the increased C3 mRNA expression associated with TNF treatment. TNF increases C3 synthesis and release from endothelial cells which were inhibited by clinical concentrations of PTX. This data further supports the potential benefit of PTX in multiple organ dysfunction and other inflammatory processes involving the endothelium by inhibiting one of the major mediators of vascular damage.
  • Fumihiko Katagiri, Hiroki Itoh, Masaharu Takeyama
    2004 年 27 巻 10 号 p. 1674-1678
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Sho-hange-ka-bukuryou-to, a traditional Chinese herbal (Kampo) medicine, has been used to treat hyperemesis of pregnancy, nausea and vomiting. Most traditional herbal medicines are prepared from several herbs. For example, Sho-hange-ka-bukuryo-to is prepared from three herbs: Pinelliae Tuber, Zingiberis Rhizoma and Hoelen. Thus, to determine the precise mechanism of the pharmacological effects of Chinese herbal medicines is too difficult. So we have elucidated the effect of some Chinese herbal medicines by examining the change of the plasma levels of brain-gut peptides. In this study, we investigated the effects of Sho-hange-ka-bukuryo-to on the plasma levels of gut-regulated peptides (gastrin, somatostatin, motilin and vasoactive intestinal peptide (VIP)) and gastrointestinal mucosa regulatory neuropeptides (calcitonin gene-related peptide (CGRP) and substance P) in healthy human subjects. A single oral administration of Sho-hange-ka-bukuryo-to caused significant increases in plasma somatostatin-, CGRP- and substance P-immunoreactive substance (IS) levels, compared with a placebo group. Transient elevation of gastrin-IS levels in the placebo group was inhibited by the administration of Sho-hange-ka-bukuryo-to, but the medicine showed no effects on plasma motilin- or VIP-IS levels. In conclusion, these results might indicate that the pharmacological action of Sho-hange-ka-bukuryo-to is closely related to changes in gastrin-, somatostatin-, CGRP- and substance P-IS levels in human plasma.
  • Fumihiko Katagiri, Shin Inoue, Yuhki Sato, Hiroki Itoh, Masaharu Takey ...
    2004 年 27 巻 10 号 p. 1679-1682
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Sho-hange-ka-bukuryo-to and Nichin-to, traditional Chinese herbal (Kampo) medicines have been used to treat vomiting and nausea. Traditional herbal medicines have frequently been used in the empirical treatment. Some patients who take these medicines have no organic disease but have conditions classified as non-ulcer dyspepsia (NUD). To determine the pharmacological effects of Sho-hange-ka-bukuryo-to, Nichin-to, and the two herbs (Pinelliae Tuber and Zingiberis Rhizoma, both of which are included in Sho-hange-ka-bukuryo-to and Nichin-to), we examined the effects of these medicines on the plasma levels of adrencorticotropic hormone (ACTH) and cortisol under stress conditions by repetitive blood sampling. After a single administration of Kampo medicine or a placebo, venous blood samples were taken before and 20—240 min after administration. A single administration of Sho-hange-ka-bukuryo-to caused significant suppression of an increase in plasma ACTH-immunoreactive substance (IS) levels at 120 to 180 min and tended to suppress increases in plasma cortisol levels at 240 min, compared with the response to a placebo. A single administration of Nichin-to caused significant suppression of increases in plasma ACTH-IS levels at 120 min compared with a placebo group, but had no effect on plasma cortisol levels. Pinelliae Tuber had no significant effects in plasma ACTH-IS or cortisol, but Zingiberis Rhizoma significantly suppressed the increase of ACTH-IS (120 min) and cortisol (180 min). These medicines have a modulatory effect on the hypothalamo-pituitary-adrenal (HPA) axis and autonomic nervous function. These effects might be beneficial in stress-related disease and suggest that this medicine has clinical pharmacological activity.
Toxicology
Regular Articles
  • Hiroe Tsutsumi, Munehiro Katagi, Mayumi Nishikawa, Hitoshi Tsuchihashi ...
    2004 年 27 巻 10 号 p. 1572-1575
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    In order to investigate the conversion of selegiline (SG), a drug used in the treatment of Parkinson's disease, to selegiline N-oxide (SGO) as a major metabolic pathway for SG, rat liver microsomal incubations were carried out in vitro in the presence of NADPH. SG was transformed into SGO in vitro as described in our previous human in vivo experiment. In the kinetic studies, the Vmax/Km value of the N-oxidation at pH 8 was found to be approximately four times greater than that at pH 7.4. The N-oxidation was also found to be inhibited by methimazole, an inhibitor of the flavin-containing monooxigenase (FMO) rather than by SKF 525A, an inhibitor of cytochrome P450s, and stimulated approximately two times by n-octylamine, an stimulator of FMO. Moreover, the N-oxidation activity remained almost unchanged in the presence of NADPH even after heating at 50 °C for a few minutes. The present data demonstrate that the N-oxidation of SG to SGO is principally mediated by FMO.
Medicinal Chemistry
Regular Articles
Notes
  • Nithyanantham Muruganantham, Ramaiah Sivakumar, Navaneetharaman Anbala ...
    2004 年 27 巻 10 号 p. 1683-1687
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    A series of 8-substituted quinolines were synthesized and tested against seizures induced by maximal electro shock (MES), pentylenetetrazole (scMet) and antihypertensive activities. Neurologic deficit was evaluated by the rotarod test. Among the newly synthesized derivatives, several compounds with a 2-hydroxypropyloxyquinoline moiety displayed excellent anticonvulsant and antihypertensive activities. Compound 20 (8-(3′-(4″-phenylpiperazino)-2′-hydroxypropyloxy)quinoline) was potent in both series as an anticonvulsive agent. 13 (8-(3′-piperazino)-2′-hydroxypropyloxyquinoline) and 14 (8-(3′-imidazolo)-2′-hydroxypropyloxyquinoline) showed very good anticonvulsant activities in the propanol series of compound, whereas in the ethane series, 1 (8-(2′-piperazino-ethanoxy)quinoline) and 2 (8-(2′-imidazolo-ethanoxy)quinoline) were the most active as anticonvulsive agents. Compounds 20 (8-(3′-(4″-phenylpiperazino)-2′-hydroxypropyloxy)quinoline), 13 (8-(3′-piperazino)-2′-hydroxypropyloxyquinoline) and 19 (8-(3′-(4″-ethylpiperazino)-2′-hydroxypropyloxy)quinoline) have shown excellent antihypertensive activity. They have significantly antagonized the pressor response elicited by adrenaline. These pharmacological results suggest that their anticonvulsant and antihypertensive effects may be correlated to the presence of β-blocking properties, and that those properties depend on the presence of aryloxypropanolamine.
Pharmacognosy
Regular Articles
  • Jang-Shim Yim, Young-Suk Kim, Sang-Kwan Moon, Ki-Ho Cho, Hyung-Sup Bae ...
    2004 年 27 巻 10 号 p. 1580-1583
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    To evaluate the pharmacological actions of herbal medicines, metabolic activities of herbal medicine components, ginsenoside Rb1, glycyrrhizin, geniposide and baicalin to their bioactive compounds compound K, 18β-glycyrrhetic acid, genipin and baicalein by fecal specimens were measured. Their metabolic activities were 646.1±591.4, 29.4±51.7, 926.3±569.6 and 3884.6±1400.1 μmol/h/g, respectively. The profiles of these metabolic activities of baicalin and ginsenoside Rb1 were not significantly different to those of water extracts of Scutellariae Radix and Ginseng Radix. None of the metabolic activities tested were different between males and females, or between ages. However, the difference in these metabolic activities in individuals was significant. These results suggest that the human intestinal microflora enzymes that convert herbal components to their bioactive compounds may be used as selection markers of responders to traditional medicines.
  • Hidenori Ueda, Kazuko Kawanishi, Masataka Moriyasu
    2004 年 27 巻 10 号 p. 1584-1587
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Caesalpinia ferrea MART. (Leguminosae) called as Jucá is one of the medicinal plants in Brazil used for diabetes. From the fruits of this plant, ellagic acid (EA) and 2-(2,3,6-trihydroxy-4-carboxyphenyl)ellagic acid (TEA) have been recently isolated as aldose reductase (AR) inhibitors. In this study, we examined to prove the inhibitory activity against AR of EA and TEA in vitro, and EA in vivo by measurement of the accumulation of sorbitol, which is the product of glucose reduction catalyzed by AR. TEA was not examined in vivo because of its shortage of yield from the fruits. EA and TEA significantly and dose-dependently inhibited sorbitol accumulation in erythrocytes, lens and sciatic nerve under incubating with glucose in vitro. EA at a dose of 75 mg/kg/d showed the most potent inhibition of sorbitol accumulation in erythrocytes, lens and sciatic nerve at 50, 75 and 100 mg/kg/d in vivo. These results suggest that the inhibitory activity of EA against AR causes to inhibit sorbitol accumulation by in vitro and in vivo experiments. EA is distributed in fruits and vegetables, so that taking them might be able to relieve diabetic complications.
  • Young-Mi Park, In-Tae Kim, Hee-Juhn Park, Jong-Won Choi, Kun-Young Par ...
    2004 年 27 巻 10 号 p. 1588-1593
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    In an attempt to find bioactive natural products with an anti-inflammatory activity, we evaluated the effects of the methanol extract of Fomes fomentarius (MEFF) on in vivo anti-inflammatory and anti-nociceptive activities. MEFF (50, 100 mg/kg/d, p.o.) reduced acute paw edema induced by carrageenin in rats, and showed MEFF analgesic activity, as determined by an acetic acid-induced writhing test and a hot plate test in mice. To investigate the mechanism of the anti-inflammatory action of MEFF, we examined the effect of MEFF on lipopolysaccharide (LPS)-induced responses in murine macrophages cell line RAW 264.7. MEFF potently inhibited the production of nitric oxide (NO), prostaglandin E2 (PGE2), and tumor necrosis factor-α (TNF-α) in LPS-stimulated RAW 264.7 macrophages. Consistent with these observations, inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) levels were reduced by MEFF in a dose-dependent manner. Furthermore, MEFF suppressed nuclear factor-κB (NF-κB) activation in LPS-stimulated RAW 264.7 macrophages. These findings suggest that the anti-inflammatory and anti-nociceptive properties of the methanol extract of MEFF may result from the inhibition of iNOS and COX-2 expression through the down-regulation of NF-κB binding activity.
  • Bo-Rim Seo, Chae-Bin Yoo, Hee-Juhn Park, Jong-Won Choi, Kyungwon Seo, ...
    2004 年 27 巻 10 号 p. 1594-1598
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    The present work was performed to investigate the effects of saucernetin-8 on proliferation and differentiation of human leukemia HL-60 cells as well as the underlying mechanisms for these effects. Saucernetin-8 exhibited a potent antiproliferative activity against HL-60 cells. This compound was also found to be a potent inducer of differentiation in human leukemia derived HL-60 cells through the examination of differentiation markers, as assessed by nitroblue tetrazolium reduction test, esterase activity assay, phagocytic activity assay, morphology change, and expression of CD14 and CD66b surface antigens. These results suggest that saucernetin-8 induces the differentiation of human leukemia cells to granulocytes and monocytes/macrophages lineage. Moreover, DNA flow-cytometry indicated that saucernetin-8 induced a G1 phase arrest of HL-60 cells. The protein and mRNA expression levels of p21 were up-regulated during saucernetin-8-dependent HL-60 cell differentiation, whereas the level of c-myc was down-regulated. Taken together, our results suggest that saucernetin-8 may have potential as a therapeutic agent in human leukemia.
  • Keiko Kawashima, Asano Nomura, Toshiaki Makino, Ken-ichi Saito, Yoshih ...
    2004 年 27 巻 10 号 p. 1599-1603
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Hange-shashin-to (HST) has been used as an herbal formula to treat inflammatory ulcerative gut diseases complicated with psychoneurosis in Japanese traditional Kampo medicine. The aim of the present study is to clarify anti-colitic effect of HST using a model of colitis induced by intracolonic instillation of 2,4,6-trinitrobenzene sulfonic acid (TNBS) in rats, and to evaluate the pharmaceutical properties of its herbal components. The colonic damage was elucidated by macroscopic damage scores, colon wet weight and area of mucosal necrosis. Orally administered HST significantly reduced the colonic damage. Other rats were orally treated with single-component berberine (BE), baicalin (BA), glycyrrhizin (GL) or saponin fraction of ginsenosides (GS), or with the mixture (TL) of BA, BE, GL and GS, or with the combinations of BA plus BE (BA-BE), or that of GL plus GS (GL-GS). Oral treatment of TL ameliorated colitis observations. However, no effects were found in the treatment of single-component BA, BE, GL or GS, whereas the GL-GS combination ameliorated the colitis. These results suggest that HST might suppress inflammatory bowel disease (IBD) and imply that there will be a potential benefit in the traditionally derived herbal combination.
  • Seong-Gyu Ko, Seung-Hee Koh, Chan-Yong Jun, Chang-Gyu Nam, Hyun-Su Bae ...
    2004 年 27 巻 10 号 p. 1604-1610
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    We performed this study to understand the molecular basis underlying the antitumor effects of Saussurea lappa, Pharbitis nil, Plantago asiatica and Taraxacum mongolicum, which have been used for herbal medicinal treatments against cancers in East Asia. We analyzed the effects of these medicinal herbs on proliferation and on expression of cell growth/apoptosis related molecules, with using an AGS gastric cancer cell line. The treatments of Saussurea lappa and Pharbitis nil dramatically reduced cell viabilities in a dose and time-dependent manner, but Plantago asiatica and Taraxacum mongolicum didn't. FACS analysis and Annexin V staining assay also showed that both Saussurea lappa and Pharbitis nil induce apoptotic cell death of AGS. Expression analyses via RT-PCR and Western blots revealed that Saussurea lappa, but not Pharbitis nil, increased expression of the p53 and its downstream effector p21Waf1, and that the both increased expression of apoptosis related Bax and cleavage of active caspase-3 protein. We also confirmed the translocation of Bax to mitochondria. Collectively, our data demonstrate that Saussurea lappa and Pharbitis nil induce growth inhibition and apoptosis of human gastric cancer cells, and these effects are correlated with down- and up-regulation of growth-regulating apoptotic and tumor suppressor genes, respectively.
  • Hideaki Matsuda, Yoshiko Kawaguchi, Miho Yamazaki, Noriko Hirata, Shun ...
    2004 年 27 巻 10 号 p. 1611-1616
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    A methanolic extract from the leaves of Piper nigrum L. showed a significant stimulatory effect on melanogenesis in cultured murine B16 melanoma cells. Activity-guided fractionation of the methanolic extract led to the isolation of two known lignans, (−)-cubebin (1) and (−)-3,4-dimethoxy-3,4-desmethylenedioxycubebin (2), together with a new lignan, (−)-3-desmethoxycubebinin (3). Among these lignans, 1 and 2 showed a significant stimulatory activity of melanogenesis without any significant effects on cell proliferation.
  • Ricardo Reyes-Chilpa, Jesús Rivera, Martha Oropeza, Pilar Mendo ...
    2004 年 27 巻 10 号 p. 1617-1620
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Hamelia patens JAQC. (Rubiaceae) is a medicinal bush widely distributed in tropical areas of the American continent. It is used in Mexican Traditional Medicine for the treatment of menstrual disorders, therefore suggesting that its chemical constituents may have some effect on myometrium contractility. Physiological effects might differ due to quantitative variations in the content of alkaloids arisisng from its wide geographical distribution. To test this hypothesis, the content of oxindole alkaloids in methanol extracts of five different samples collected in Mexico was quantified by GC-MS. Each extract was assayed on contractility of estrogen-primed rat myometrium. Variations in the content of alkaloids were observed among the different samples. All samples relaxed in a concentration-dependent manner the high KCl-induced contraction in rat myometrium. Those which lack rumberine and/or maruquine displayed a higher relaxant effect than samples containing them, suggesting that these alkaloids might counteract the effects of isopteropodine. However, in contrast with verapamil, Hamelia patens metanol extracts are poor relaxants.
  • Satoru Mitsuma, Eriko Ishigaki, Ryuji Sugiyama, Tetsuya Asamizu, Kyoji ...
    2004 年 27 巻 10 号 p. 1621-1625
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Transgenic sesame (Sesamum schinzianum ASCH.) was produced by Agrobacterium-mediated transfection of a carrot calmodulin gene, cam-4, which was specifically expressed upon the contact of carrot cells with oligogalacturonide elicitor. Coding region of cam-4 was ligated to the downstream of 35S promoter of cauliflower mosaic virus and subcloned into pMATGBO-DB3.1. A. tumefaciens 4404 was transformed with the constructed vector, and the crown gall tissues formed in the sesame seedlings were transferred onto appropriate media to obtain the re-differentiated plants. The reverse-transcription polymerase chain reaction followed by Southern blot analysis revealed that cam-4 gene was appreciably expressed in the transgenic plants. Activities of two key enzyme regulating phenylpropanoid metabolisms, phenylalanine ammonia-lyase and caffeic acid O-methyltransferase, and the contents of phenolic compounds in the transformed sesame were markedly elevated as compared with those of the control. These results suggest that the over-expression of cam-4 gene enhances the biosynthetic activities of phenylpropane derivatives in the transformed sesame plants.
Notes
Biopharmacy
Regular Articles
  • Yaw-Bin Huang, Yi-Hung Tsai, Wan-Chiech Yang, Jui-Sheng Chang, Pao-Chu ...
    2004 年 27 巻 10 号 p. 1626-1629
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    The purpose of this study was to develop propranolol extended release formulations containing hydroxypropylmethylcellulose (HPMC). The results indicate that the drug release from the tablet form containing a high amount of HPMC was incomplete, and avicel addition could increase the release percent at a later stage. In order to readily obtain an optimal formulation, response surface methodology and multiple response optimization utilizing a quadratic polynomial equation was used. The model formulations were prepared according to a factorial design. The effects of causal factors including the HPMC/drug ratio (X1) and avicel level (X2), on drug release were also measured. The drug release percentage at 1.5, 4, 8, 14 and 24 h were the target response and were restricted to not more than 25%, 35—50%, 55—70%, 75—90%, and 95—110%, respectively. The results showed that the optimized formulation provided a dissolution pattern equivalent to the predicted curve, which indicated that the optimal formulation could be obtained using response surface methodology. The mechanism of drug release from HMPC matrices tablets followed quasi-Fickian diffusion.
  • M. Zahirul I. Khan, Dragica Raušl, Ružica Zanoški, S ...
    2004 年 27 巻 10 号 p. 1630-1635
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Loratadine was studied both in vitro and in vivo (in healthy humans) to classify it according to the Biopharmaceutics Classification System (BCS) in order to gain more understanding of the reasons for its highly variable nature with respect to plasma time profiles, and to determine the most appropriate dissolution test conditions for in vitro assessment of the release profile of the drug from solid dose forms. Based on the solubility of loratadine determined under various pH conditions and its permeability through Caco-2 monolayers, loratadine was classified as a Class II drug. Plasma profiles were predicted by convolution analysis using dissolution profiles obtained under various pH and hydrodynamic conditions as the input function and plasma time data obtained from a syrup formulation as the weighting function. The predicted profiles based on dissolution studies done at gastric pH values were in reasonable agreement with the mean bio-data suggesting dissolution testing should be done at gastric pH values. However, the bio-data were highly variable and it is suggested this may be due, at least in part, to high individual gastric pH variability and dissolution occurring in the intestine on some occasions, and therefore, dissolution testing should also be done in simulated intestinal fluid.
  • Kap Joo Park, Hye Yun Kim, Byung Joon Chang, Hyung Hoan Lee
    2004 年 27 巻 10 号 p. 1636-1641
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    This study sought to investigate the ameliorating effects of soy 11S protein on the impacts of alcohol consumption in rat hepatocytes and in reducing total cholesterol levels and total lipid levels in the serum. Liver histology and the clinically important enzyme markers (Aspartate Aminotransferase: AST and Alanine Aminotransferase: ALT) of rats, administered with both alcohol and soy 11S protein treatments, were compared with those in the control group. The treatment regimen (11S soy protein extract) significantly reduced serum ALT and AST levels, indicating the hepato-protective effects of soy 11S protein. Furthermore, total cholesterol and total lipid levels were significantly reduced. In addition to preventing the presence of lipid droplets and secondary lysosomes, electron microscopy indicated that the administration of the soy 11S protein treatment preserved important hepatocyte structures. These results indicate that soy 11S protein can positively mediate the effects of alcohol on hepatocytes and general liver functions.
  • Masato Taguchi, Takashi Nozawa, Kouichi Mizumaki, Hiroshi Inoue, Katsu ...
    2004 年 27 巻 10 号 p. 1642-1648
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    This study was performed to estimate the mean pharmacokinetic parameters of routinely administered metoprolol in middle-aged and elderly Japanese patients. Whole blood concentration data (65 samples) at steady-state following repetitive administration to 34 patients were analyzed using a nonlinear mixed effects model. A one-compartment model was parameterized in terms of oral clearance (CL/F) and apparent volume of distribution (V/F). We evaluated the effect of polymorphic alleles (CYP2D6*2, CYP2D6*10, CYP2C19*2 and CYP2C19*3), age, gender, and heart failure on the pharmacokinetic parameters of metoprolol. The CL/F value in patients homozygous for the CYP2D6*10 allele was 64% lower than that in patients with a CYP2D6*1/*1 or *1/*2 genotype. The CL/F value in older (>70 years old) patients was 26% lower than that in younger (≤70 years old) patients. In addition, the V/F value in patients homozygous for the CYP2D6*10 allele was 25% lower than that in patients with the CYP2D6*1/*1 or *1/*2 genotype. On the other hand, the CYP2C19 genotype, gender, and heart failure showed no significant effects on the pharmacokinetics of metoprolol. The results suggest that the pharmacokinetic variability of metoprolol in Japanese extensive metabolizers of CYP2D6 is very large, probably because CYP2D6*10 is responsible not only for the decreased systemic clearance (CL) but also for the increased bioavailability (F) of the drug.
  • Michiya Kobayashi, Hiroshi Saitoh, Shujiro Seo, Veronika Butterweck, S ...
    2004 年 27 巻 10 号 p. 1649-1652
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    We investigated the effect of Apocynum venetum L. extract (AV) on the activity of cytochrome P450 (CYP) 3A and P-glycoprotein (P-gp). The plasma concentration of nifedipine (NF), which is a substrate for CYP3A, did not change after oral administration with AV (3.3 mg/kg). Also, AV (3.3 and 33 mg/kg) did not affect the intestinal absorption of NF. In the rats treated with multiple administrations (15 mg/kg/d) of St. John's wort extract (SJW) for 2 weeks, the plasma concentration of NF after oral administration was significantly decreased. On the other hand, there was no significant differences in the pharmacokinetic parameters of NF between AV-treated (3.3 mg/kg/d) and none-treated rats. Furthermore, the intestinal absorption of methylprednisolone, which is a substrate for P-gp, was not affected by AV treatment for 2 weeks. These results suggest that, unlike SJW, the recommended dose of AV (3.3 mg/kg/d) would not influence hepatic CYP3A and intestinal P-gp in rats.
  • Teruko Nishio, Masato Maruyama, Toyokazu Yoshida, Yoshihiko Watanabe, ...
    2004 年 27 巻 10 号 p. 1653-1655
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    We have investigated the secretion polarity of human interferon-β (HuIFN-β) exogenously expressed in the porcine renal proximal tubule cell line, LLC-PK1. In these cells, stably expressed HuIFN-β was secreted to the apical and basolateral sides. However, when transiently expressed by apical lipofection, HuIFN-β was secreted to both cell sides, while basolateral-preferential secretion was seen for basal transfection. Confocal imaging using enhanced green fluorescent protein (EGFP)-tagged HuIFN-β revealed no difference in the subcellular distribution in either of the chimeric protein-expressing cells examined. These results suggest that the secretion polarity of HuIFN-β is regulated by a post trans-Golgi network in a cell type-dependent manner.
Notes
  • Fumio Itagaki, Masato Homma, Kohji Takara, Noriaki Ohnishi, Teruyoshi ...
    2004 年 27 巻 10 号 p. 1694-1696
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    The aim of our study was to investigate the effects of rabeprazole, a proton pump inhibitor, on MDR1 expressed on human colon carcinoma cell line, Caco-2, and MDR1-overexpressing human cervical carcinoma cell line, HeLa cells selected by exposure to 100 nM vinblastine (Hvr100-6 cells). Inhibitory effects of rabeprazole on MDR1-mediated transport of Rhodamine123 were examined in these cells. A thousand micro molar rabeprazole increased Rhodamine 123 uptakes in Caco-2 and Hvr100-6 cells by 68% and 185%, respectively. No significant effects of rabeprazole were observed at the concentration of 1—100 μM. Since rabeprazole did not show any effects on Rhodamine 123 transport via MDR1 at the plasma levels (approximately 1 μM), it was considered that the drug interaction with MDR1 substrates would be minimal even though the interaction occurred in the patients with rabeprazole treatment.
  • Ryu Hirayama, Koyo Nishida, Shintaro Fumoto, Mikiro Nakashima, Hitoshi ...
    2004 年 27 巻 10 号 p. 1697-1699
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    The present study was undertaken to investigate liver site-specific gene transfer following the administration of naked plasmid DNA (pDNA) to the liver surface in mice. We examined whether genes could be delivered to the liver site specifically by utilizing the glass-made diffusion cell that is able to limit the contact dimension between the liver surface and pDNA solution administered. Gene expression was detected at the site of diffusion cell attachment (site 1) and was significantly higher than in other liver sites and tissues. Moreover, gene expression was also detected at deeper site from the liver surface (noncontact side with pDNA solution). The level of gene expression at site 1 did not change significantly with pDNA treatment for 10, 30, and 60 min. In conclusion, we demonstrated that naked pDNA administered to the liver surface in mice was taken up from its surface, and subsequently the protein encoded by pDNA could be produced site specifically.
Miscellaneous
Regular Articles
  • Gi-Young Kim, Myung-Geum Han, Yung-Sun Song, Byung-Cheul Shin, Yong-Il ...
    2004 年 27 巻 10 号 p. 1656-1662
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    Mushroom polysaccharides are increasingly being utilized to treat a wide variety of diseases. Phellinus linteus proteoglycan (PL) has been reported to have anti-tumor and immunomodulatory properties. However, the cellular and molecular mechanism underlying its therapeutic effect is poorly understood. In this study, we investigated whether PL induces the phenotypic and functional maturation of murine bone marrow-derived dendritic cells (DC) and the possibility that Toll-like receptors (TLRs), which are known to be involved in immune-related responses, may be the receptor(s) of PL. The expression of surface molecules, including major histocompatibility complex (MHC) class II and CD86, increased on DC that were stimulated in a dose-dependent manner with PL, in comparison with unstimulated DC. Furthermore, PL increases the production of IL-12 by DC, as well as the IL-2 secretion and proliferation of allogeneic T cells. In addition, the activities of PL on DC were significantly reduced by treating the cells with anti-TLR2 or anti-TLR4 antibody (Ab) prior to PL, suggesting that both of them are possible receptors of PL. Also, maturation of DC by PL was able to directly activate mitogen-activated protein kinases (MAPKs), such as ERK1/2 and p38, and the nuclear transcription factor NF-κB p65. Also, the pretreatment of DC with inhibitors of NF-κB p65, and ERK and p38 MAPK signal pathways inhibited PL-induced up-regulation of surface molecules, such as MHC class II and CD86, and IL-12 production. Our results demonstrated that PL stimulation could induce the phenotypic and functional maturation of DC via TLR2 and/or TLR4 mediated-NF-κB, ERK and p38 MAPK signal pathways.
  • Noriko Tagawa, Kimie Muraoka, Yoshihiro Okamoto, Mikio Nishida, Masana ...
    2004 年 27 巻 10 号 p. 1663-1665
    発行日: 2004年
    公開日: 2004/10/01
    ジャーナル フリー
    In our previous study, we have investigated the serum levels of dehydroepiandrosterone (DHEA) in type II collagen (CII)-induced arthritis (CIA) mice. During the study, we found that in normal control mice, serum levels of DHEA in the latter half of the experimental period (13—16 weeks old) were significantly lower than those at the beginning of the experiment (10 weeks old). However, in CIA mice, such decreases were not observed by CII treatment. To examine the cause of the retention of DHEA during the development of arthritis in CIA mice in this study, 17α-hydroxylase/C17-20 lyase P450 (CYP17) mRNA expressions were measured by real time RT-PCR and the CYP17 enzyme activities were investigated in the liver and testis on days 6, 13, 28 and 48 after CII treatment in DBA/1J mice. There were no significant differences of CYP17 expressions in the liver between control and CIA mice at each experimental day, while a significant increase of expression in the testis of CIA mice was observed on day 48. On the other hand, CYP17 enzyme activities on days 28 and 48 in testis microsome (Mc) from the CIA mice were significantly higher than those of the control on the same day, while no significant differences of activities in liver Mc were observed between the CIA and control mice. These findings suggested that the cause of the retention of DHEA on days 28 and 48 after CII treatment may be the increase of CYP17 expression and the enzyme activities in the testis.
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