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Seungheon Lee, Dong Hyun Kim, Ji Hye Lee, Eun Seong Ko, Won Bo Oh, Yon ...
2013 年 36 巻 11 号 p.
1692-1699
発行日: 2013/11/01
公開日: 2013/11/01
[早期公開] 公開日: 2013/08/22
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The aim of this study was to identify the effects of 85% methanolic extract of
Morus alba leaves (EMA), which is a traditional herb, in mice. The effects of EMA on the anxiolytic-like behaviour were studied using the elevated plus maze (EPM) and hole-board test. To elucidate the mode of action of the anxiolytic-like effects of EMA, the mice were subjected to the co-administration of EMA (200 mg/kg,
per os (
p.o.)) and either antagonist. EMA (at 200 or 400 mg/kg) significantly increased the percentages of time-spent in the open arms and entries into the open arms of the EPM
versus vehicle-treated control group (
p<0.05). Moreover, in the hole-board test, EMA (200 and 400 mg/kg) significantly increased the number of head-dips
versus vehicle-treated control group (
p<0.05). However, there were no changes in the locomotor activity and myorelaxant effects in any group compared with the vehicle-treated control group. In addition, the anxiolytic-like effects of EMA were abolished by thioperamide (10 mg/kg, intraperitoneally (i.p.)), which is a histamine H
3 receptor antagonist. Moreover, results from reverse transcription polymerase chain reaction (RT-PCR) also revealed that the amygdalal histidine decarboxylase mRNA expression levels in EMA (200 mg/kg)-treated group were significantly higher than those in the vehicle-treated controls (
p<0.05). These results suggest that EMA might prove to be an effective anxiolytic agent and that EMA acts
via the histaminergic system in central nerve system.
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Cheng-Gang Liu, Xiao-Li Wang, Xiao-Wei Du, De-You Jiang, Nai-Zhi Geng, ...
2013 年 36 巻 11 号 p.
1700-1707
発行日: 2013/11/01
公開日: 2013/11/01
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Metabolomics is a new platform based on the comprehensive analysis of low molecular weight metabolites and provides a powerful approach to discover biomarkers in biological systems. Modified Sinisan (MSNS), a traditional Chinese medicine formula, displayed bright prospects in the prevention and therapy of liver injury. However, its molecular mechanism of hepatoprotective effects remains unclear. This paper was designed to explore the effects and potential mechanisms of MSNS against dimethylnitrosamine-induced liver injury. Global metabolic profiling was performed by ultra-performance liquid chromatography/electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC/ESI-Q-TOF-MS) in conjunction with multivariate data analysis and pathway analysis. Eleven serum biomarkers were identified and pathway analysis results showed that phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism, tryptophan metabolism, retinol metabolism, tyrosine metabolism were perturbed by liver injury. More importantly, MSNS has showed satisfactory pharmacological effect on liver injury through partially regulating the perturbed pathways, correlates well to the biochemical and histopathological detection results. The present study proved that the robust metabolomics approach is promising for unraveling hepatoprotective effects of MSNS and these findings provide new insights into mechanisms of the liver injury, and its pathophysiologic processes.
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Tatsuya Morimoto, Yoichi Sunagawa, Yasufumi Katanasaka, Sae Hirano, Ma ...
2013 年 36 巻 11 号 p.
1708-1714
発行日: 2013/11/01
公開日: 2013/11/01
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Curcumin has various biological activities including antioxidant and antiinflammatory actions, and alcohol detoxification. However, because of its poor absorption efficiency, it is difficult for orally administered curcumin to reach blood levels sufficient to realize its bioactivities. We have generated capsules and tablets containing Theracurmin, a highly absorptive curcumin. In addition, we recently created a drinkable preparation of Theracurmin. To evaluate the absorption efficiency of this type of curcumin, we performed a single-dose, double-blind, 4-way crossover study. We compared plasma curcumin levels after the administration of Theracurmin beverage and 3 other drinkable types of curcumin sold in Japan. Twenty-four healthy subjects (male/female=13/11, age: 23–32) were administered with these 4 drinkable preparations of curcumin. The area under the blood concentration–time curve at 0–8 h was found to be 1.5 to 4.0-fold higher with Theracurmin than with the other 3 kinds of curcumin beverage. Moreover, maximal plasma curcumin concentrations (0–8 h) of Theracurmin were 1.8 to 3.8 times higher than those of the other 3 curcumin beverages. These data indicate that our newly prepared Theracurmin beverage exhibits a much better absorption efficiency than other kinds of curcumin beverage sold in Japan.
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Shigehiro Matsunaga, Noboru Uchide, Midori Shono, Kunio Ohyama, Makoto ...
2013 年 36 巻 11 号 p.
1715-1721
発行日: 2013/11/01
公開日: 2013/11/01
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Human cytomegalovirus (CMV) is the most common cause of viral intrauterine infection in developed countries. It has been shown that CMV DNA was frequently detected in the fetal membranes when the placenta was infected
in utero. However, it is still not clear whether CMV replicates in constituent cells of the fetal membranes. We investigated CMV infection of primary cultured chorion and amnion cells prepared from human fetal membrane tissues. In both types of cell cultures, rounded cells were observed at day 8 and 12 after CMV inoculation, and virus yields in culture supernatants were increased after the inoculation. In both types of cells, viral immediately early 1 (IE1) protein-positive nuclei were scattered at day 4 after the inoculation, and IE1 mRNA was expressed throughout day 1 to 12 after CMV inoculation. In chorion cell cultures, the number of IE1 protein-positive nuclei increased significantly at day 8 and 12 after CMV inoculation as compared to day 4, by which foci were formed. Furthermore, an evident increase in levels of lactate dehydrogenase leakage from chorion cells was observed after CMV inoculation. Contrary, these phenomena were not observed in amnion cell cultures. These results demonstrated that both chorion and amnion cells were permissive to CMV infection, while the velocity of cell-to-cell spread of CMV infection in amnion cells was much lower than that in chorion cells. Therefore, the present study suggests that CMV may replicate rapidly in the chorion cell layer and slowly in the amnion cell layer during intrauterine infection.
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Yu Inoue, Seiji Hasegawa, Takaaki Yamada, Yasushi Date, Hiroshi Mizuta ...
2013 年 36 巻 11 号 p.
1722-1730
発行日: 2013/11/01
公開日: 2013/11/01
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Hydroquinone (HQ) is a chemical compound that inhibits the functions of melanocytes and has long been known for its skin-whitening effect. According to previous studies, the Tyrosinase (Tyr) activity inhibitory effect and melanocyte-specific cell toxicity are known depigmenting mechanisms; however, details of the underlying mechanisms are unknown. Arbutin (Arb) is also known for its Tyr activity inhibitory effect and is commonly used as a skin-whitening agent. However, the detailed depigmenting mechanism of Arb is also not yet fully understood. Few studies have attempted to elucidate the effects of HQ and Arb on undifferentiated melanocytes. In this study, we examined the effects of HQ and Arb throughout each stage of differentiation of melanocytes using a mouse embryonic stem cell (ESC) culture system to induce melanocytes. The results showed that HQ in particular downregulated the early stage of differentiation, in which neural crest cells were generated, and the late stage of differentiation, in which melanogenesis became active. On the other hand, Arb had no effect on the differentiation of melanocytes, and only suppressed melanogenesis by specifically suppressing elevations in Tyr expression in the late stage of differentiation.
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Yoshimasa Takafuji, Yuriko Higuchi, Atsushi Muro, Kohei Oshiro, Shiger ...
2013 年 36 巻 11 号 p.
1731-1738
発行日: 2013/11/01
公開日: 2013/11/01
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Artificial introduction of functional molecules on the cell surface may be a promising way to improve the therapeutic effects of cell therapy. Pegylated lipids are conventionally used in drug carriers. The lipid part of pegylated lipids noncovalently interacts with the cell surface. However, little information is available regarding conditions for cell-surface modification by using pegylated lipids. In this study, we synthesized fluorescein-labeled pegylated lipids and evaluated the factors that affect modification efficiency by using human mesenchymal stem cells (hMSCs). As the concentration of the pegylated lipid as well as the exposure time increased, the modification efficiency increased. The modification efficiency at 37°C was 20- and 3-fold higher than that at 4°C and 25°C, respectively. In addition, with an increase in the molecular weight of polyethylene glycol (PEG), more pegylated lipids were extracellularly distributed than those intracellularly distributed. At the optimal condition, pegylated lipids were observed mainly on the cell membrane by confocal microscopy. In contrast, the cell condition (adherent or nonadherent) had little or no effect on the cell-surface modification efficiency. The results of this study will be useful for constructing an optimal modification method for introducing functional molecules on the cell surface.
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Yuya Yoshida, Takumi Tsuji, Sayaka Watanabe, Ayane Matsushima, Yuki Ma ...
2013 年 36 巻 11 号 p.
1739-1746
発行日: 2013/11/01
公開日: 2013/11/01
[早期公開] 公開日: 2013/08/30
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Fingolimod (FTY720) is known to have a significant therapeutic effect in various autoimmune disease models. Here, we examined FTY720 in a model of rheumatoid arthritis, induced by immunizing DBA/1 mice with a peptide consisting of residues 325 through 339 of glucose-6-phosphate isomerase (GPI
325–339). The efficacy was evaluated in terms of macroscopic findings, inflammatory cell infiltration and autoantibody level. Prophylactic administration of FTY720 from the day of immunization significantly suppressed the development of paw swelling, but therapeutic administration of FTY720 from onset of symptoms on day 8–9 was less effective. Interestingly, however, combination treatment with FTY720 plus GPI
325–339 for 5 d after onset of symptoms significantly reduced the severity of symptoms in all mice, and no relapse occurred after booster immunization. Taking into account the reported mechanism of action of FTY720, these results indicate that combination treatment with FTY720 plus pathogenic autoantigen might efficiently induce immune tolerance by sequestering circulating autoantigen-specific lymphocytes from blood and peripheral tissues to the secondary lymphoid tissues. Combination treatment with FTY720 plus pathogenic autoantigen may become a breakthrough treatment for remission-induction in patients with autoimmune diseases including rheumatoid arthritis.
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Myoung-Yun Pyo, Bo-kyung Park, Jeong June Choi, Mihi Yang, Hyun Ok Yan ...
2013 年 36 巻 11 号 p.
1747-1753
発行日: 2013/11/01
公開日: 2013/11/01
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The oral consumption of capsicum has been reported to increase interleukin (IL)-2 and interferon (IFN)-γ production in Peyer’s patches (PP); however, the active components responsible for these effects have not been completely identified. The beneficial biological effects of green peppers cultivated under environmentally friendly farming conditions (ECP), without the use of chemical pesticides, have rarely been compared with those of green peppers cultivated under conventional farming conditions (CCP). Oral administration of ECP extract significantly induced the production of IL-2 and IFN-γ in concanavalin A-treated cells from PP
ex vivo; their levels were much higher than those in the CCP extract-treated group. A comparative analysis of the HPLC profiles indicated a 1.7-fold increase of a peak, named EF-1, at 415 nm in the ECP extract. The major component of EF-1 was identified as pheophytin
a, which is a chlorophyll
a molecule lacking a central Mg
2+ ion, as determined from NMR data. Intake of pheophytin
a and chlorophyll
a significantly increased IL-2 and IFN-γ production, and the percentage of IL-2- and IFN-γ-producing CD4+ T-cells in PP. Taken together, our data suggest that ECPs produce a higher content of pheophytin
a than CCPs, and pheophytin
a and chlorophyll
a are immune-modulating components in green vegetables.
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Su Ui Lee, Hyun Ju In, Mi So Kwon, Bi-oh Park, Minmi Jo, Mun-Ock Kim, ...
2013 年 36 巻 11 号 p.
1754-1759
発行日: 2013/11/01
公開日: 2013/11/01
[早期公開] 公開日: 2013/08/29
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G-protein coupled receptor 43 (GPR43) serves as a receptor for short-chain fatty acids (SCFAs), implicated in neutrophil migration and inflammatory cytokine production. However, the intracellular signaling pathway mediating GPR43 signaling remains unclear. Here, we show that β-arrestin 2 mediates the internalization of GPR43 by agonist. Agonism of GPR43 reduced the phosphorylation and nuclear translocation of nuclear factor-κB (NF-κB), which was relieved by short interfering RNA (siRNA) of β-arrestin 2. Subsequently, mRNA expression of proinflammatory cytokines, interleukin (IL)-6 and IL-1β, was downregulated by activation of GPR43 and knockdown of β-arrestin 2 recovered the expression of the cytokines. Taken together, these results suggest that GPR43 may be a plausible target for a variety of inflammatory diseases.
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Ryo Morofuji, Tomohiro Hikima, Kakuji Tojo
2013 年 36 巻 11 号 p.
1760-1765
発行日: 2013/11/01
公開日: 2013/11/01
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Skin has various types of transporters and is a biochemically active organ. These aspects of skin influence the distribution of chemicals in skin and their elimination from skin. The biochemical and histological variations of the skin must be taken into account when conducting transdermal penetration research. Here we used hairless mouse skin to investigate the percutaneous absorption of chemicals
in vitro from the stratum corneum (SC) side to the viable skin (VS) side (forward direction) and from the VS side to the SC side (backward direction). We examined the effects of molecular weight, lipophilicity (Log
K
o/w), electric charge, and the molecular structure of penetrants. The penetration flux of verapamil hydrochloride (VRP) for the backward direction was 3.2 times larger than that for the forward direction. The flux values of benzoic acid (BA) and
para-hydroxybenzoic acid (
pHBA) for the forward direction were 2.1 and 4.6 times larger than those for the backward direction, respectively. This directional difference was caused by the active transporter for VRP, the histological distribution of BA solubility, and the intermolecular hydrogen bonding between
pHBA and skin tissue in the stripped skin. Across intact skin, in contrast, there was no difference in the skin penetration profile between the forward direction and backward directions.
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Daisuke Aibara, Kimihiko Matsusue, Kohei Matsuo, Soichi Takiguchi, Fra ...
2013 年 36 巻 11 号 p.
1766-1772
発行日: 2013/11/01
公開日: 2013/11/01
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Fat-specific protein 27 gene (
FSP27), isolated by screening for genes specifically expressed in fully differentiated mouse adipocytes, belongs to the cell death-inducing DNA fragmentation factor, alpha subunit-like effector family. FSP27 is induced in not only adipose tissue but also the liver of
ob/
ob mice, and it promotes the development of fatty liver. The
FSP27 gene is expressed in a fatty liver-specific manner and is not detected in the normal mouse liver.
FSP27 expression is directly regulated by the induction of the hepatic peroxisome proliferator-activated receptor γ (PPARγ) in
ob/
ob fatty liver. In the present study, expression of hepatic
FSP27 mRNA was determined in non-genetic fatty liver models. The
FSP27 gene was markedly induced in the high-fat- or methionine- and choline-deficient (MCD) diet-induced fatty liver, but it was not elevated in alcohol-induced fatty liver. Interestingly, the induction of
FSP27 mRNA due to the MCD diet was independent of PPARγ levels and completely absent in the liver from PPARγ-null mice. These results suggest that
FSP27 mRNA expression in the liver depends on the etiology of fatty liver.
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Xingyu Zhao, Lianhai Jin, Nan Shen, Bin Xu, Wei Zhang, Hongli Zhu, Zhe ...
2013 年 36 巻 11 号 p.
1773-1778
発行日: 2013/11/01
公開日: 2013/11/01
[早期公開] 公開日: 2013/08/22
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Salidroside, a phenylpropanoid glycoside isolated from
Rhodiola rosea L., shows potent antioxidant property. Herein, we investigated the protective effects of salidroside against hydrogen peroxide (H
2O
2)-induced oxidative damage in human endothelial cells (EVC-304). EVC-304 cells were incubated in the presence or absence of low steady states of H
2O
2 (3–4 µ
M) generated by glucose oxidase (GOX) with or without salidroside. 3(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH) assays were performed, together with Hoechst 33258 staining and flow cytometric analysis using Annexin-V and propidium iodide (PI) label. The results indicated that salidroside pretreatment attenuated endogenous H
2O
2 induced apoptotic cell death in EVC-304 cells in a dose-dependent pattern. Furthermore, Western blot data revealed that salidroside inhibited activation of caspase-3, 9 and cleavage of poly(ADP-ribose) polymerase (PARP) induced by endogenous H
2O
2. It also decreased the expression of Bax and rescued the balance of pro- and anti-apoptotic proteins. All these results demonstrated that salidroside may present a potential therapy for oxidative stress in cardiovascular and cerebrovascular diseases.
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Sung Chul Kwak, Cheol Lee, Ju-Young Kim, Hyun Mee Oh, Hong-Seob So, My ...
2013 年 36 巻 11 号 p.
1779-1786
発行日: 2013/11/01
公開日: 2013/11/01
[早期公開] 公開日: 2013/08/28
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Excessive osteoclastic bone resorption plays a critical role in inflammation-induced bone loss such as rheumatoid arthritis and periodontal bone erosion. Therefore, identification of osteoclast targeted-agents may be a therapeutic approach to the treatment of pathological bone loss. In this study, we isolated chlorogenic acid (CGA) from fructus of
Gardenia jasminoides to discover anti-bone resorptive agents. CGA is a polyphenol with anti-inflammatory and anti-oxidant activities, however, its effects on osteoclast differentiation is unknown. Thus, we investigated the effect of CGA in receptor activator of nuclear factor-kappa B (NF-κB) ligand (RANKL)-induced osteoclast differentiation and RANKL signaling. CGA dose-dependently inhibited RANKL-mediated osteoclast differentiation in bone marrow macrophages (BMMs) without any evidence of cytotoxicity. CGA inhibited the phosphorylation of p38, Akt, extracellular signal-regulated kinase (ERK), and inhibitor of nuclear factor-kappa B (IκB), and IκB degradation by RANKL treatment. CGA suppressed the mRNA expression of nuclear factor of activated T cells c1 (
NFATc1),
TRAP and
OSCAR in RANKL-treated bone marrow macrophages (BMMs). Also, overexpression of
NFATc1 in BMMs blocked the inhibitory effect of CGA on RANKL-mediated osteoclast differentiation. Furthermore, to evaluate the effects of CGA
in vivo, lipopolysaccharide (LPS)-induced bone erosion study was carried out. CGA remarkably attenuated LPS-induced bone loss based on micro-computed tomography and histologic analysis of femurs. Taken together, our findings suggest that CGA may be a potential treatment option for osteoclast-related diseases with inflammatory bone destruction.
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Seo-Yeon Yoon, Ji-Hee Yeo, Seung-Dae Han, Dong-Jun Bong, Beomsoo Oh, D ...
2013 年 36 巻 11 号 p.
1787-1793
発行日: 2013/11/01
公開日: 2013/11/01
[早期公開] 公開日: 2013/08/29
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Oxaliplatin, which is used as one of anti-cancer drugs, commonly induces peripheral neuropathic pain. We have previously reported that an injection of diluted bee venom (DBV) produced a significant anti-nociceptive effects in several pain models of mice or rats. In this study, we evaluated time- and dose-dependent development of oxaliplatin-induced mechanical allodynia in bilateral hind paws of mice, and investigated the effect of DBV injection on this mechanical allodynia. DBV (0.1 mg/kg) was subcutaneously injected into the Zusanli acupoint 2 weeks after oxaliplatin (10 mg/kg) injection. One hour after DBV injection, we observed a significant reduction of mechanical allodynia in the ipsilateral hind paw, but not in the contralateral hind paw to DBV injection site. We subsequently examined whether this effect of DBV was related to the activation of peripheral nerves in DBV injected site, and then whether it was mediated by the activation of spinal cord alpha-2 adrenoceptors or opioid receptors. Subcutaneous pre-injection of 2% lidocaine (40 mg/kg) into the Zusanli acupoint completely blocked the anti-allodynic effect of DBV. Intrathecal pretreatment with yohimbine (25 µg/mouse), an alpha-2 adrenoceptor antagonist, also prevented the anti-allodynic effect of DBV, whereas pretreatment with naloxone (20 µg/mouse), an opioid receptor antagonist, did not block the effect of DBV. Taken together, these findings demonstrate that DBV injection into the Zusanli acupoint significantly reduces ipsilateral mechanical allodynia generated by oxaliplatin in mice, and also suggest that this anti-allodynic effect is dependent on the peripheral nerve activation in injected site and spinal cord alpha-2 adrenoceptors.
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Kosuke Kanda, Yukinobu Kodama, Tomoaki Kurosaki, Masanobu Imamura, Hir ...
2013 年 36 巻 11 号 p.
1794-1799
発行日: 2013/11/01
公開日: 2013/11/01
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The purpose of the present study was to investigate the usefulness of the ternary complex with protamine and γ-polyglutamic acid (γ-PGA), which are biodegradable materials for foods and medical products, as a safe gene delivery vector. We formed cationic binary complexes (plasmid DNA (pDNA)/protamine complexes) with high transfection efficiency. The binary complex showed slight toxicity probably related to its total cationic charge. Then, we formed ternary complexes (pDNA/protamine/γ-PGA complexes) by addition of anionic polymer, γ-PGA, and they showed no cytotoxicity. The transfection efficiency of the pDNA/protamine/γ-PGA complexes was as high as that of the pDNA/protamine complexes, although their zeta potentials were different. Inhibition study of the gene expressions in B16-F10 cells suggested that pDNA/protamine complexes were taken up by caveolae-mediated endocytosis and macropinocytosis. On the other hand, pDNA/protamine/γ-PGA complexes were taken up by clathrin-mediated endocytosis and macropinocytosis. Thus, we succeeded in developing the ternary complex as a safe gene delivery vector with biocompatible materials.
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Tomoaki Kurosaki, Yukinobu Kodama, Takahiro Muro, Norihide Higuchi, Ta ...
2013 年 36 巻 11 号 p.
1800-1806
発行日: 2013/11/01
公開日: 2013/11/01
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In this experiment, we developed a novel safe and effective gene delivery vector coated with γ-polyglutamic acid (γ-PGA-coated complexes). The γ-PGA-coated complex was composed of chiseled spherical nano-particles with anionic charges. The plasmid DNA/polyethyleneimine complex (non-coated complex) showed high transgene efficiency in the spleen and lung after intravenous administration in mice, with high liver toxicity and lethality. On the other hand, γ-PGA-coated complex selectively showed high transgene efficiency in the spleen without such toxicity. Furthermore, the γ-PGA-coated complex highly accumulated and showed high gene expression in the marginal zone of the spleen. Those results strongly indicated that γ-PGA-coated complex was suitable as a DNA vaccine vector. We therefore applied γ-PGA-coated complex to melanoma DNA vaccine, pUb-M. The γ-PGA-coated complex containing pUb-M significantly inhibited the growth and metastasis of a melanoma cell line, B16-F10 cells. In conclusion, we developed a splenic gene vector, γ-PGA-coated complex, as a novel technology for clinical vaccination.
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Naoki Yoshikawa, Shintaro Fumoto, Mikiro Nakashima, Kenta Shimokawa, H ...
2013 年 36 巻 11 号 p.
1807-1813
発行日: 2013/11/01
公開日: 2013/11/01
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We analyzed the effect of serum and fibronectin on pulmonary transgene expression after intravenous injection of cationic liposome–plasmid DNA (pDNA) complex (lipoplex) in mice. 1,2-Dioleoyl-3-trimethylammonium-propane (DOTAP) methyl sulfate salt/cholesterol lipoplex was incubated with several serum components for 5 min at 37°C prior to injection. We analyzed pulmonary transgene expression and pulmonary accumulation of lipoplex. While interaction with serum did not decrease pulmonary transgene expression, interaction with heat-inactivated serum did decrease it. Moreover, interaction with fibronectin enhanced pulmonary transgene expression. Inhibition of the binding of fibronectin to integrin decreased pulmonary transgene expression after injection of untreated lipoplex. We found that pulmonary accumulation of lipoplex changed depending on the kind of interacting serum components after injection. Furthermore, interaction with fibronectin increased pulmonary accumulation of lipoplex. Interaction with serum was required for pulmonary gene transfer following intravenous injection of lipoplex. Fibronectin appears to be a particularly critical component. Furthermore, the binding of fibronectin interacting with lipoplex to integrin was an important mechanism for pulmonary transgene expression.
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Miwa Uesugi, Mio Hosokawa, Haruka Shinke, Emina Hashimoto, Tamotsu Tak ...
2013 年 36 巻 11 号 p.
1814-1821
発行日: 2013/11/01
公開日: 2013/11/01
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Association between cytochrome P450 (CYP)
3A4*1G genotype of donors (
n=412) and/or recipients (
n=410), and the pharmacokinetics of tacrolimus and the risk of acute cellular rejection was examined in Japanese living-donor liver transplant patients between 2004 and 2011. The concentration/dose (C/D) ratio of tacrolimus in patients carrying graft liver with
CYP3A4*1/*1 was significantly higher during 7 d after surgery than in that with
CYP3A4*1/*1G (214
vs. 157 [ng/mL]/[mg/kg/day],
p<0.01). After postoperative day 8, no significant difference was observed among
CYP3A4*1G genotypes in the graft liver. However, the C/D ratio in
CYP3A4*1/*1 of the intestine was significantly higher than that in
CYP3A4*1G/*1G for 5 weeks after surgery (postoperative days 1–14;
p<0.001, postoperative days 15–35;
p<0.01). During postoperative days 14 and 26, acute cellular rejection incidences tended to be lower in the patients with graft liver carrying the
CYP3A4*1/*
1 allele than in the patients carrying
CYP3A4*1G allele (8.7%
vs. 14.6%,
p=0.0973). However,
CYP3A4*1G in the intestine had almost no effect on the incidence of rejection (9.9% in
CYP3A4*1/*1 vs. 12.5% in
CYP3A4*1G allele,
p=0.4824).
CYP3A4*1G was significantly related to mRNA expression of CYP3A5 rather than of CYP3A4 in the graft liver and intestine and was strongly linked with the
CYP3A5*1. Thus, we elucidated that
CYP3A4*1G genotype in the intestine was an important indicator of the pharmacokinetics of tacrolimus, whereas this genotype in the graft liver tended to influence the frequency of acute cellular rejection after transplantation.
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Takuro Kobori, Shinichi Harada, Kazuo Nakamoto, Shogo Tokuyama
2013 年 36 巻 11 号 p.
1822-1828
発行日: 2013/11/01
公開日: 2013/11/01
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Previously, we reported that repeated oral administration of etoposide (ETP) increases P-glycoprotein (P-gp) expression in association with activation of ezrin/radixin/moesin (ERM) in the small intestine. Radixin has recently attracted attention for its critical role in the plasma membrane localization of certain drug transporters including P-gp by working as a scaffold protein. However, there have been no report investigating that radixin really interacts with small intestinal P-gp and is involved in the mechanism by which the levels of P-gp are altered. Here, we examined whether radixin is involved in the increased P-gp expression in the small intestine after ETP treatment. Repeated oral treatment with ETP (10 mg/kg/day) for 7 d significantly increased ERM proteins bound to P-gp in the small intestine as determined by immunoprecipitation analysis. In particular, radixin but not ezrin or moesin bound to P-gp was dramatically increased in association with the up-regulation of P-gp in the small intestinal membrane, and radixin was highly co-localized with P-gp as measured by immunofluorescence analysis. In conclusion, radixin may contribute, at least in part, to an increase in the expression of the small intestinal P-gp upon induction with repeated oral treatment with ETP.
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Kentaro Kono, Atsuyoshi Okada, Atsuko Ishikawa, Tetsuya Aiba
2013 年 36 巻 11 号 p.
1829-1834
発行日: 2013/11/01
公開日: 2013/11/01
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To clarify whether peripheral inflammation has a remote effect on the central nervous system, the electrolyte disposition between the circulating blood and central nervous system was evaluated in rats with carrageenan-induced acute peripheral inflammation (API). λ-Carrageenan was subcutaneously injected in the hind paw of the rat, and lithium was utilized as a surrogate marker of sodium. When the plasma and cerebrospinal fluid (CSF) concentrations of lithium were examined following lithium being intravenously administered, it was revealed that the CSF concentration of lithium in API rats is reduced compared to that in normal rats, while the plasma concentration profile of lithium in API rats is indistinguishable from that in normal rats. The pharmacokinetic analysis showed that the lithium disposition from the plasma to CSF markedly decreased by 35.8% in API rats compared to that in normal rats. On the other hand, when lithium was immediately administered into the lateral ventricle, its elimination profiles in CSF were not different between normal and API rats. It is therefore probable that the lithium disposition from the plasma to CSF alters in API rats, reflecting the entry process of electrolytes from the circulating blood to brain tissue being suppressed in response to peripheral inflammation.
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Yuka Sone, Ryosuke Nakamura, Hidemitsu Pan-Hou, Tomoo Itoh, Masako Kiy ...
2013 年 36 巻 11 号 p.
1835-1841
発行日: 2013/11/01
公開日: 2013/11/01
[早期公開] 公開日: 2013/08/28
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The characteristics of bacteria take up mercury into cells
via membrane potential-dependent sequence-divergent members of the mercuric ion (Mer) superfamily,
i.e., a periplasmic mercuric ion scavenging protein (MerP) and one or more inner membrane-spanning proteins (MerC, MerE, MerF, and MerT), which transport mercuric ions into the cytoplasm, have been applied in engineering of bioreactor used for mercurial bioremediation. We engineered bacteria to express MerC, MerE, MerF, or MerT with or without MerP to clarify their individual role and potential in transport of mercurial. By immunoblot analysis using specific polyclonal antibody, the proteins encoded by
merC,
merE,
merF,
merT or
merP, were certainly expressed and identified in the membrane fraction. Bacteria expressing MerC, MerE, MerF or MerT in the absence of MerP transported significantly more C
6H
5Hg(I) and Hg(II) across bacterial membrane than their isogenic strain.
In vivo expression of MerP in the presence of all the transporters did not cause apparent difference to the C
6H
5Hg(I) transport, but gives an apparently higher Hg(II) transport than that did by MerE, MerF or MerT but not by MerC. Among the four transporters studied, MerC showed more potential to transport Hg(II) across bacterial membrane than MerE, MerF and MerT. Together these findings, we demonstrated for the first time that in addition to MerE and MerT, MerF and MerC are broad-spectrum mercury transporters that mediate both Hg(II) and phenylmercury transport into cells. Our results suggested that MerC is the most efficient tool for designing mercurial bioremediation systems, because MerC is sufficient for mercurial transport into cells.
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Amr Selim Abu Lila, Masako Ichihara, Taro Shimizu, Tatsuhiro Ishida, H ...
2013 年 36 巻 11 号 p.
1842-1848
発行日: 2013/11/01
公開日: 2013/11/01
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We have reported that PEGylated liposomes lose their long-circulating properties when injected twice into the same animal within a certain interval (the accelerated blood clearance (ABC) phenomenon). We assumed that this phenomenon was triggered
via the abundant secretion of anti-polyethylene glycol (PEG) immunoglobulin M (IgM) in response to the first dose of PEGylated liposomes and that the spleen played an important role in the production of anti-PEG IgM. However, no direct evidence has yet confirmed this suspicion. In the current study, we verified, both
in vitro and
ex vivo, that spleen cells are indeed responsible for the production of anti-PEG IgM in response to PEGylated liposomes. In this study, spleen cells obtained from either naïve mice or mice pre-treated with PEGylated liposomes induced the production of anti-PEG IgM in a dose- and time-dependent manner, upon incubation with PEGylated liposomes. In addition, we confirmed that among the different fractions of splenic B cells, IgM-positive B cells, rather than CD45R-positive or CD19-positive splenic B cells, which are presumed to be the marginal zone B (MZB) cells, are the major cells producing anti-PEG IgM in the response to stimulation by PEGylated liposomes. These results may provide new insights into the mechanisms underlying the anti-PEG IgM production in response to the stimulation by PEGylated liposomes.
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Tomoya Sasahara, Katsutoshi Yayama, Hiroshi Okamoto
2013 年 36 巻 11 号 p.
1849-1856
発行日: 2013/11/01
公開日: 2013/11/01
[早期公開] 公開日: 2013/08/23
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Hyperosmotic stress induces the contractile response of vascular smooth muscle cells (VSMCs). Previous studies have demonstrated that cytoskeleton reorganization and Rho/Rho-kinase-mediated inactivation of myosin light chain phosphatase (MLCP) play an important role in hyperosmotic vasoconstriction, but the precise mechanism is unknown. This study aimed to investigate the contractile response of endothelium-denuded rings of rat aortas to hyperosmolar sucrose (160 m
M) in the presence or absence of inhibitors for various protein kinases. We found that the hyperosmotic constriction of aortic rings was attenuated not only by ML-7 or hydroxyfasudil, specific inhibitor for myosin light chain kinase (MLCK) or Rho-kinase, respectively, but also by SB203580, a specific inhibitor for p38 mitogen-activated kinase (p38 MAPK). Hyperosmolar sucrose evoked a transient increase in cytosolic free Ca
2+ in rat VSMCs, and this response was not affected by SB203580. Western blot analysis of proteins extracted from rings showed that the hyperosmolar sucrose stimulated phosphorylation of the Rho-kinase-mediated myosin phosphatase target subunit 1, myosin light chain (MLC), and p38 MAPK. The experiments performed using a combination of the kinase inhibitors showed that hyperosmolarity-induced MLC phosphorylation is partially mediated
via the SB203580-sensitive pathway and is independent of both MLCK and Rho-kinase-mediated inactivation of MLCP. Furthermore, the hyperosmolarity-induced increase in the F-actin/G-actin ratio in rings was attenuated not only by hydroxyfasudil but also by SB203580. These results suggest that p38 MAPK is involved in hyperosmotic vasoconstriction
via stimulation of MLC phosphorylation and cytoskeleton reorganization through pathways independent of activation of MLCK and/or Rho-kinase-mediated mechanisms.
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