Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
30 巻 , 2 号
選択された号の論文の38件中1~38を表示しています
Regular Articles
  • Nataliya Doronicheva, Hiroyuki Yasui, Hiromu Sakurai
    2007 年 30 巻 2 号 p. 213-217
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    The antioxidative activity of flavonoids depends upon a combination of many factors, such as the concentration and chemical structure of the flavonoids and the arrangement of functional groups in their structure. In the present study, to evaluate the antioxidative effect of several types of flavonoids on catalase activity at a physiological H2O2 concentration, a chemiluminescent (CL) method was used. The H2O2/luminol-dependent CL intensity in a system containing 3.7 nM catalase and low concentrations (10—100 nM) of green tea flavanols (epigallocatechin gallate; EGCG and epicatechin gallate; EG) was enhanced in comparison with that of a system without catalase, suggesting that EGCG and EG partially suppressed catalase activity. On the other hand, flavone and flavonols such as rutin (a 3-glycosidic flavone), quercitrin (a 3-glycosidic flavonol), myricetin, and kaempferol (flavonols), respectively, lowered the CL intensity to a greater extent at low concentrations (<0.1 μM) when catalase was present than when catalase was absent, indicating that these flavonoids activate catalase. In addition, isoflavone and flavanone such as daidzein and naringenin, respectively, exhibited weak antioxidative activities against H2O2 without any effect on the catalase activity over a wide range of flavonoid concentrations (0.04—0.4 μM). From these results, it was for the first time suggested that the binding of flavonoids to the heme moiety or a protein region of catalase contributes to the enhancement of catalase activity.
Regular Articles
  • Haruhiko Kamada, Takayuki Okamoto, Maki Kawamura, Hiroko Shibata, Yasu ...
    2007 年 30 巻 2 号 p. 218-223
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Many biologically active proteins need to be delivered intracellularly to exert their therapeutic action inside the cytoplasm. Cell penetrating peptides (CPPs) have been developed to efficiently deliver a wide variety of cargo in a fully biological active form into a range of cell types for the treatment of multiple preclinical disease models. To further develop this methodology, we established a systematic approach to identify novel CPPs using phage display technology. Firstly, we screened a phage peptide library for peptides that bound to the cell membrane. Secondly, to assess functionality as intracellular carriers, we recombined cDNAs of binding peptides with protein synthesis inhibitory factor (PSIF) to create fusion proteins. Randomly chosen clones were cultured and expression of peptide-PSIF fusion proteins induced, followed by screening of protein synthesis activity in cells. Using this systematic approach, novel and effective CPPs were rapidly identified. We suggest that these novel cell-penetrating peptides can utilized as drug delivery tools for protein therapy or an analytical tool to study mechanisms of protein transduction into the cytoplasm.
  • Shinya Abe, Satoko Suyama, Seiko Chiba, Hajime Nakao, Yosuke Ohtake, Y ...
    2007 年 30 巻 2 号 p. 224-229
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    We have recently reported that transferrin (Tf)-unbound gallium-67 (67Ga) may be taken up into the liver of carbon tetrachloride (CCl4)-treated rats. In the present study, we attempted to clarify detailed mechanism of Tf-unbound 67Ga uptake by hepatocytes treated with CCl4 using in vitro experimental system. Hepatotoxic damages by CCl4 are mostly attributed to radical formed by an action of cytochrome P450. P450 isozymes have a higher expression in the perivenous hepatocytes (PVH) more than periportal hepatocytes (PPH). Therefore, we thought that the uptake of 67Ga which had been used for the detection of liver damage might have a zonal difference. The results of ALT activities showed that the CCl4 exposure for 4 h strongly impaired PVH more than PPH. The uptake of 67Ga by PVH treated with CCl4 was also higher than that by PPH. Moreover, the uptake of 45Ca by PVH was higher than that by PPH. In order to investigate whether 67Ga passed through calcium channel of hepatocytes, we made use of calcium channel blocker and activator. The Ca2+-channel blocker, verapamil, significantly decreased the uptakes of 45Ca and 67Ga by PPH and PVH pretreated with CCl4. The addition of the Ca2+-channel activator, Bay K8644, significantly increased the uptake both of 45Ca and 67Ga by PPH pretreated with CCl4. In the present study, it was demonstrated that the uptake of Tf-unbound 67Ga preferentially occurred in CCl4-damaged PVH and 67Ga was taken up into the hepatocytes in part through calcium channel.
  • Kunihiko Gyoubu, and Mitsuo Miyazawa
    2007 年 30 巻 2 号 p. 230-233
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    The in vitro metabolism of (−)-camphor was examined in human liver microsomes and recombinant enzymes. Biotransformation of (−)-camphor was investigated by gas chromatography-mass spectrometry (GC-MS). (−)-Camphor was oxidized to 5-exo-hydroxyfenchone by human liver microsomal cytochrome (P450) enzymes. The formation of metabolites of (−)-camphor was determined by the relative abundance of mass fragments and retention time on gas chromatography (GC). CYP2A6 was the major enzyme involved in the hydroxylation of (−)-camphor by human liver microsomes, based on the following lines of evidence. First, of eleven recombinant human P450 enzymes tested, CYP2A6 catalyzed the oxidation of (−)-camphor. Second, oxidation of (−)-camphor was inhibited by (+)-menthofuran and anti-CYP2A6 antibody. Finally, there was a good correlation between CYP2A6 contents and (−)-camphor hydroxylation activities in liver microsomes of 9 human samples.
  • Mi-Ja Kim, Kyung Hee Jung, Yoon Kyung Uhm, Kang-Hyun Leem, Hye Kyung K ...
    2007 年 30 巻 2 号 p. 234-236
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Oxidative stress is produced under diabetic conditions and involved in progression of pancreatic β-cell dysfunction. Both an increase in reactive oxygen free radical species (ROS) and a decrease in the antioxidant defense mechanism lead to the increase in oxidative stress in diabetes. Electrolyzed reduced water (ERW) with ROS scavenging ability may have a potential effect on diabetic animals, a model for high oxidative stress. Therefore, the present study examined the possible anti-diabetic effect of ERW in genetically diabetic mouse strain C57BL/6J-db/db (db/db). ERW with ROS scavenging ability reduced the blood glucose concentration, increased blood insulin level, improved glucose tolerance and preserved β-cell mass in db/db mice. The present data suggest that ERW may protects β-cell damage and would be useful for antidiabetic agent.
  • Yoshitaka Kanayama, Chizuru Aoki, Yasuo Sakai
    2007 年 30 巻 2 号 p. 237-241
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    The basic concept of tissue engineering, an actively applied technology in regenerative medicine, is the coordination between cells, scaffolds, and proliferation factors. Collagen and gelatin are materials that have mostly been used as scaffolds because of their excellent affinity for cells, they are biodegradable, and their ability to form gel. However, very few of the collagen or gelatin preparations used as tissue engineering material have been prepared with low levels of endotoxin, which may trigger fever, shock and a fall in blood pressure even in very minute quantities. Therefore we decided to decrease the content of endotoxin in gelatin so that this biomaterial is not harmful to the body but still maintains its properties, namely, bioaffinity and biodegradation for use in the fields of cell culture and regenerative medicine. In this paper, we describe a method whereby endotoxin could be removed from gelatin solution by using an ultrafiltration membrane with a molecular weight cut-off of 100 kDa and low endotoxin gelatin, endotoxin activity in 0.1% gelatin solution was below 0.03 EU/ml, was obtained as the resulting filtrate solution.
Regular Articles
  • Eun Kyung Kim#, Kang Beom Kwon#, Ju Hyung Lee, Byung Hyun Park, Jin Wo ...
    2007 年 30 巻 2 号 p. 242-246
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Cytokines produced by immune cells infiltrating pancreatic islets are important mediators of β-cell destruction in insulin-dependent diabetes mellitus. Scoparone (6,7-dimethoxycoumarin) is known to have a wide range of pharmacological properties in vitro. In this study, the effects of scoparone on cytokine-induced β-cell dysfunction were examined. Presence of scoparone significantly protected interleukin-1β (IL-1β) and interferon-γ (IFN-γ)-mediated cytotoxicity of RINm5F, a rat insulinoma cell line, and preserved glucose-stimulated insulin secretion in rat pancreatic islets. Scoparone also resulted in a significant reduction in IL-1β and IFN-γ-induced nitric oxide (NO) production, a finding that correlated well with reduced levels of the inducible form of NO synthase (iNOS) mRNA and protein. The molecular mechanism by which scoparone inhibited iNOS gene expression appeared to involve the inhibition of NF-κB activation. These results revealed the possible therapeutic value of scoparone for the prevention of diabetes mellitus progression.
  • Jie Qiu, Hai-Qing Gao, Rui-Hai Zhou, Ying Liang, Xu-Hua Zhang, Xu-Ping ...
    2007 年 30 巻 2 号 p. 247-253
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Digitalis has been used to treat congestive heart failure for more than 200 years, although the dual effects (proliferation and death) induced by digitalis on cell growth have been known for many years, the mechanisms by which digitalis causes the actions were not completely known. The aim of this work was to characterize the proliferative effect of ouabain on cell growth in endothelial cells, and, to do the differential proteomic analysis of human umbilical vein endothelial cells (HUVEC) in response to ouabain and examine changes in protein expression. HUVEC were exposed to different concentrations (0.1—100 nM) of ouabain at 12—48 h intervals. Cell growth and morphological changes of HUVEC treated with ouabain were compared with cells under nontreated conditions. Ouabain stimulated HUVEC cell proliferation at low concentrations and induced cell death at higher concentrations. Using proteomics study, we identified 32 proteins of HUVEC with various important cellular functions and revealed 8 proteins such as Annexin A1, Annexin A2, Malate dehydrogenase, Myosin regulatory light chain 2 (MRLC2), Profilin-1, S100 calcium-binding protein A13, Triosephosphate isomerase and Translationally controlled tumor protein, regulated by low-dose ouabain treatment and MRLC2 was subsequently confirmed by Western blot. Our results give new insights into the cellular and molecular mechanisms of the proliferation action of low-dose ouabain on HUVEC and provide new avenues for the treatment of cardiovascular diseases.
  • Dan Li, Li-Jun Wu, Shin-ichi Tashiro, Satoshi Onodera, Takashi Ikejima
    2007 年 30 巻 2 号 p. 254-260
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Oridonin, an active component isolated from the plant Rabdosia rubescens, has been reported to exhibit antitumor effects, but little is known about its molecular mechanism of action. In this study, we first investigated the mechanism involved in oridonin-induced cell death in human epidermoid carcinoma A431 cells, which overexpress epidermal growth factor receptor (EGFR). After treatment with various doses of oridonin for 24 h, the majority of A431 cells underwent apoptosis in a time- and dose-dependent manner as measured by an LDH activity-based assay. Treatment with oridonin at various concentrations for 24 h caused significant inhibition on the total tyrosine kinase activities and downregulation of EGFR expression or EGFR phosphorylation. Oridonin significantly affected the localization of EGFR and phosphorylated EGFR on the cell membrane. However, genistein (a well-known tyrosine kinase inhibitor) did not induce apoptotic A431 cell death. Importantly, oridonin exhibited much stronger inhibitory effect on the total tyrosine kinase activities or EGFR tyrosine phosphorylation as well as much stronger suppression on EGFR and phosphorylated EGFR localization than genistein in A431 cells. Taken together, oridonin exerted a potential inhibitory effect on the tyrosine kinase activity of A431 cells. The decrease in the tyrosine kinase activity and the blockage of EGFR tyrosine phosphorylation might be one of the causes of oridonin-induced A431 cell death.
  • Yohko Yamazaki, Setsuko Kunimoto, Daishiro Ikeda
    2007 年 30 巻 2 号 p. 261-265
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Hypoxia is a common feature of many solid tumors and contributes to their progression. Hypoxic cells in the tumor are not only involved in therapeutic resistance to chemotherapy and radiotherapy but are also relevant to tumor angiogenesis. To identify novel hypoxia-selective cytotoxins, we screened 20000 cultured broths of microorganisms and found that rakicidin A showed significant hypoxia-selective cytotoxicity. Rakicidin A was approximately 17.5-fold more cytotoxic under hypoxic than under normoxic conditions. CoCl2 and antioxidants had no effect on the rakicidin A cytotoxicity under normoxic conditions and rakicidin A did not show the inhibitory effects on HIF-1 transcriptional activity under hypoxic conditions. Thus, although the action mechanism of the hypoxia-selective cytotoxicity of rakicidin A was unknown, our screening study suggested that rakicidin A acts as an antitumor agent for selective therapy against solid tumors.
  • Takako Iso, Kazunobu Futami, Teruaki Iwamoto, Yasuhiro Furuichi
    2007 年 30 巻 2 号 p. 266-271
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    We report that the expression of Bloom helicase (BLM) was up-regulated by 17β-estradiol (E2) in estrogen receptor (ER)-positive mammary tumor MCF-7 cells, but was hardly modulated in ER-negative mammary tumor MDA-MB-231 cells. ER antagonist ICI182780 blocked the E2 effect on BLM expression in MCF-7 cells. From these results we conclude that ER participates in up-regulation of BLM expression in MCF-7 cells by means of E2. Similar results were obtained when MCF-7 cells were treated with bisphenol A (BPA), an endocrine-disrupting chemical having a weak estrogenic activity. The ER binding ability of BPA is estimated at 1/1000 of E2 ability, and in this study about 1000-times more BPA was needed for the same levels of estrogenic effect of E2. The expression of cell-cycle associated genes, cdc6, MCM5, MCM2, Myt1, PCNA and AuroraA were up-regulated by E2 and BPA treatment in MCF-7 cells accompanied by up-regulation of BLM. In this BLM promoter study, Sp1 elements in the upper region of BLM modulated transcription, but were not indispensable for E2 response. Our results suggested that up-regulation of BLM expression by E2 and BPA is ER-dependent and may be responsible for repair of DNA damage caused by the genotoxicity of these estrogenic agents.
  • Takaki Tosaki, Hikaru Sakamoto, Jun Kitahara, Hirotaka Imai, Yasuhito ...
    2007 年 30 巻 2 号 p. 272-278
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    The synthesis of platelet-activating factor (PAF) by human umbilical vein endothelial cell (HUVEC) in response to H2O2 was significantly increased in a concentration-dependent manner. When HUVEC were pretreated with diethyl maleate, which depletes intracellular glutathione, PAF synthesis was enhanced 3-fold upon 5 mM H2O2-treatment. Intracellular redox was involved in regulating PAF synthesis, since the addition of antioxidants such as N-acetylcysteine, pyrrolidinecarbodithioic acid (PDTC), and Trolox reduced PAF production in H2O2-treated HUVEC. The activity of acetyl-CoA: 1-O-alkyl-2-lyso-sn-glycero-3-phosphocholine acetyltransferase, which is involved in the last step of PAF synthesis, was also activated in H2O2-treated cells. However, exogenous lyso-PAF addition had not effected to acetyltransferase activity. The acetyltransferase activity responded quickly to H2O2-treatment, but the activation was transitory. A tyrosine kinase inhibitor and a calmodulin antagonist blocked acetyltransferase activity in H2O2-stimulated cells, suggesting that tyrosine kinase and calcium/calmodulin-dependent protein kinase are involved in regulating acetyltransferase activity. These observations suggest that H2O2 is one of the modulators of lyso-PAF acetyltransferase activity via a phosphorylation system and platelet-activating factor (PAF) synthesis.
  • Hua Zhou, Xi Yang, Nai-li Wang, Ya-ou Zhang, Guo-ping Cai
    2007 年 30 巻 2 号 p. 279-283
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    The newly identified fat cell-secreted factor, visfatin, is insulin-mimetic and play a positive role in attenuating insulin resistance and diabetes. Natural steroidal saponins including tigogenin saponins have been reported to provide anti-diabetic activity and modulate glucose metabolism, but the mechanism remains unknown. In this study, we examined the effect of macrostemonoside A, a tigogenin steroidal saponin isolated from the bulbs of Allium macrostemon Bung on the expression of visfatin in differentiated 3T3-L1 adipocytes. It was found that macrostemonoside A markedly enhanced the synthesis and secretion of visfatin protein in 3T3-L1 adipocytes, and increased visfatin mRNA in a dose and time dependent manner as well. Moreover, visfatin promoter-driven luciferase expression in cells was elevated by macrostemonoside A, which was blocked by SB-203580, a specific inhibitor of p38 MAPK pathway. Lastly, we found that macrostemonoside A did not affect the expression of PPARγ and its DNA-binding ability to visfatin promoter. These results indicate that macrostemonoside A could potently stimulate visfatin expression in 3T3-L1 adipocytes, which occurs at the transcriptional level and is mediated at least partially via p38 MAPK signaling pathway. Its regulation on visfatin in adipocytes may constitute an important element in its improvement of insulin resistance and diabetes.
Notes
  • Takeshi Kinoshita, Hiroyoshi Ariga, Ken-ichi Matsumoto
    2007 年 30 巻 2 号 p. 354-358
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    We developed an easy and fast method to isolate extracellular matrix tenascin-X (TNX) from various tissues in mice based on TNX antibody affinity purification. We purified approximately 350-kDa cellular interstitial TNX (iTNX) from the spleen, liver and kidney as well as 200-kDa serum TNX (sTNX). Since the nature and significance of glycosylation in TNX remains to be elucidated, glycobiochemical properties of purified TNX were characterized by lectin blot analysis. Lectin blots by Con A, LCA, PHA-E4, RCA120 or WGA revealed the presence of N-glycan in the cellular TNX and especially complex-type N-glycan in the serum TNX. In addition, the iTNX from liver and kidney also possessed O-glycan based on the reaction to PNA. The binding to AAL indicated that iTNX from the three tissues possesses fucose linked α1,6 to a pentasaccharide core, whereas sTNX does not. The reaction to SSA but not to MAM suggested the presence of sialic acid linked α2,6 to galactose in both cellular and serum TNX. Lectin blots of trypsin-treated iTNX from the spleen also demonstrated that WGA alone reacts to the t300 product derived from the amino-terminal 300-kDa portion.
  • Naoyuki Okita, Yuki Kudo, Sei-ichi Tanuma
    2007 年 30 巻 2 号 p. 359-362
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Checkpoint kinase 1 (Chk1) plays important roles in genotoxic stress-induced cell cycle checkpoint and in normal cell cycle progression. Here, we show that Chk1 is cleaved in the treatment of apoptotic dose of etoposide (ETP) or cisplatin (CIS) but not of viable dose in HeLa S3 cells. The cleavage of Chk1 was completely inhibited by an irreversible and cell-permeable pan-caspase inhibitor, N-benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethylketone (z-VAD-fmk). These results identify Chk1 as a novel substrate that is cleaved by a caspase-dependent manner during genotoxic stress-induced apoptosis. Our data may also indicate the existence of a novel Chk1-regulated apoptotic pathway.
Regular Articles
  • Ayako Ogasawara, Nami Komaki, Hitoshi Akai, Kumiko Hori, Hiromi Watana ...
    2007 年 30 巻 2 号 p. 284-286
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    When mucin was added to Candida albicans under hyphal growth conditions, the hyphal formation was inhibited. After the 24 h incubation, the ratio of hyphal cells was 95.7±1.13% in the absence of mucin and no hyphal cells were observed in the presence of 1000 μg/ml mucin. The ratio of hyphal cells began to decreases at 6 h in the mucin addition group. Although mucin has antifungal activity, the concentration of mucin used in this assay did not inhibit the growth of C. albicans, indicating that the inhibition of hyphal formation was not due to the inhibition of germination by its antifungal activity. Expression of RAS1mRNA in C. albicans was inhibited by mucin. These results suggest that the inhibition of hyphal formation by mucin was caused by interruption of the hyphal formation signal of C. albicans.
  • Kumiko Horiuchi, Sumiko Shiota, Teruo Kuroda, Tsutomu Hatano, Takashi ...
    2007 年 30 巻 2 号 p. 287-290
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    We found that a crude extract from Salvia officinalis (sage) reduced the minimum inhibitory concentrations (MICs) of aminoglycosides in vancomycin-resistant enterococci (VRE). We isolated the effective compound from the extract and identified it as carnosol, one of diterpenoids. Carnosol showed a weak antimicrobial activity, and greatly reduced the MICs of various aminoglycosides (potentiated the antimicrobial activity of aminoglycosides) and some other types of antimicrobial agents in VRE. Carnosic acid, a related compound, showed the similar activity. The effect of carnosol and carnosic acid with gentamicin was synergistic.
Regular Articles
  • Sachiko Tanaka, Toshihiko Hirano, Toyokazu Saito, Nobuo Wakata, Kitaro ...
    2007 年 30 巻 2 号 p. 291-296
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Tacrolimus hydrate (FK506) reduces the symptoms of myasthenia gravis (MG) due to its immunosuppressive properties. A drug efflux pump P-glycoprotein (P-gp) actively transports FK506 out of target cells, thereby reducing their efficacy. We investigated the influence of FK506 therapy on the P-gp function of peripheral-blood mononuclear cells (PBMCs) in MG patients. Six MG patients treated with FK506 (MG(FK+)), four MG patients treated without FK506 administration (MG(FK−)), and 18 healthy subjects were included in this study. P-gp function was estimated by transporter activity that was inferred from a decrease in fluorescent P-gp substrate Rhodamine 123 (Rh123) and its inhibition by cyclosporine A (CsA). The P-gp efflux function in MG (FK+) patients assessed by the Kolmogorov–Smirnov (KS) statistic D was lower than in the healthy subjects (p=0.0084). However, PBMC sensitivity to FK506 in MG (FK+) patients was significantly higher compared to that of the healthy subjects (p=0.02). There was a significant correlation between the Rh123 efflux activity and PBMC sensitivity to FK506 in vitro (p=0.011). The data raise the possibility that FK506 treatment attenuated P-gp function in the PBMCs of the MG patients.
  • Yu-Mi Yang, Jun-Mo Chung, Hyewhon Rhim
    2007 年 30 巻 2 号 p. 297-302
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Many behavior studies indicate that cholecystokinin (CCK) is related to nociception and anxiety/panic actions in the midbrain periaqueductal gray (PAG). We previously reported that a sulfated form of CCK octapeptide (CCK-8S) produced excitatory effects at both pre- and postsynaptic loci in PAG neurons using slice preparations and whole-cell patch-clamp recordings. Here, we further examined the detailed mechanism of CCK-8S in acutely isolated PAG neurons of the rat using fura-2-based imaging of intracellular Ca2+ concentration ([Ca2+]i) and whole-cell patch-clamp recordings. Application of 1 μM CCK-8S produced an increase of [Ca2+]i, and its effect did not desensitize. This CCK-8S-induced [Ca2+]i increase was inhibited by the CCK2 receptor antagonist L-365260 but not by the CCK1 receptor antagonist L-364718. In addition, the effect of CCK-8S was eliminated by removing extracellular Ca2+, but not by an addition of the intracellular Ca2+ reuptake inhibitor thapsigargin. When simultaneous recordings of [Ca2+]i imaging and whole-cell patch-clamp were performed, CCK-8S-induced [Ca2+]i increase was significantly reduced at a membrane holding potential of −60 mV while CCK-8S-induced inward current was still observed. Current-voltage plots revealed that CCK-8S-induced inward current reversed near the equilibrium potential for K+ ions with a decreased membrane conductance. However, CCK-8S produced a significant inhibition on high-voltage-activated Ca2+ channel currents. These results suggest that CCK-8S can excite PAG neurons by inhibiting K+ channels, and CCK-8S-induced [Ca2+]i increase occurs secondary to depolarization. The evidence presented here expands our understanding of cellular mechanisms for CCK-mediated anti-analgesic and anxiogenic actions in the PAG.
  • Yoshinori Funakami, Taeko Hata, Eiji Itoh, Seigo Itano
    2007 年 30 巻 2 号 p. 303-308
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Rats stressed by specific alternation of rhythm in temperature (SART) show various symptoms of disautonomia, increased pulse rates, continuous hypotension, and severe orthostatic hypotension (OH) when they are subjected to postural change. The OH symptoms are improved by muscarinic M2-receptor blockers. In the present study, effects of β-adrenoceptor blocking agents on OH in SART-stressed rats were investigated. Anesthetized rats were restrained on a board in the supine position, and direct blood pressure and ECG were measured automatically using Fluclet® Jr.2. Postural change was performed by raising the rat's head up to a 60° angle for 4 min. Unstressed rats treated with hexamethonium showed large decrease in blood pressure, small reflex from the bottom of pressure and decreased tachycardia reflex, whereas isoproterenol showed little changes. In SART-stressed rats, isoproterenol alleviated the decrease in blood pressure in postural change, brought large reflex from the bottom of pressure and increased tachycardia reflex, whereas hexamethonium had little changes. Propranolol and atenolol induced the similar changes as those seen by hexamethonium. ICI-118,551, a selective β2-adrenoceptor antagonist showed large reflex from the bottom of pressure and increased tachycardia reflex in stressed rats, whereas little changes in unstressed rats. In conclusion, it was suggested that the hypotension in OH manifestation time of rats reflects the state of peripheral blood vessels, and β1-adrenoceptors played a role in compensatory tachycardia reflex and β2-adrenoceptors in blood pressure reflex. The circulatory regulation in SART-stressed rats seems to be poorly functioning in nervous reflex in postural changes.
  • Hai-Yan Peng, Jun-Rong Du, Guang-Yi Zhang, Xi Kuang, Yan-Xin Liu, Zhon ...
    2007 年 30 巻 2 号 p. 309-312
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    The present study investigated the effect of Z-Ligustilide (LIG), a characterized 3-n-alkyphthalide derivative existed in many medical Umbelliferae plants, on permanent focal ischemic brain injury in rats. Focal cerebral ischemia was induced by the occlusion of middle cerebral artery (MCA) for 24 h. LIG (20, or 80 mg/kg), orally administered at 2 h after ischemia, reduced the cerebral infarct volumes by 48.29% and 84.87% respectively compared to control group as visualized by 2,3,5-triphenyltetrazolium chloride (TTC) staining (p<0.01). Treatment with LIG could dose-dependently reduce brain swelling by 68.62% and 82.08% (p<0.01), and significantly improve behavioral deficits (p<0.01). In addition, LIG at the above used doses had no significant effect on rat body temperature. These data, along with previous findings in our lab demonstrating the neuroprotective effects of LIG in transient cerebral ischemia, suggest that LIG may be a potential neuroprotective agent for the treatment of ischemic stroke in future.
  • Takahisa Noto, Yasuhisa Furuichi, Masayuki Ishiye, Nobuya Matsuoka, Ic ...
    2007 年 30 巻 2 号 p. 313-317
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    We investigated the neuroprotective effect of tacrolimus (FK506) on the ischemia-reperfusion injury caused by transient focal brain ischemia induced by middle cerebral artery (MCA) occlusion for 60 min in rats. Neuronal damage visualized as a decrease of MAP2 immunoreactivity was observed in the cerebral cortex at 9 h after MCA occlusion and further expanded at 24 h. Hypoxic areas visualized with an immunohistochemical reaction for 2-nitroimidazole, a hypoxia marker (hypoxyprobe-1), and accumulation of granulocytes and platelets were also observed at 9 h and 24 h after MCA occlusion. Tacrolimus (1 mg/kg, i.v.), administered immediately after MCA occlusion, attenuated cortical damage and decreased the hypoxyprobe-1 positive area, as well as the number of granulocytes and platelets at 24 h after MCA occlusion. Immunohistochemical analysis showed that tacrolimus reduced the number of blood vessels positively stained for ICAM-1, E-selectin and P-selection. These results suggested that tacrolimus limited attachment of granulocytes and platelets to blood vessels by inhibiting the expression of adhesion molecules and protected neuronal tissue from hypoxic insults.
  • Yoshihiko Chiba, Yingyan Yu, Hiroyasu Sakai, Miwa Misawa
    2007 年 30 巻 2 号 p. 318-323
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Although an involvement of matrix metalloproteinase (MMP)-12 in the development of chronic obstructive pulmonary disease (COPD) and airway inflammation has been suggested, its detailed role in the airways is not well known now. In the present study, the changes in the expression and localization of MMP-12 in airways of repeatedly antigen-challenged rats were investigated to show an association of MMP-12 with allergic bronchial asthma. Rats sensitized by dinitrophenylated Ascaris antigen were 3 times repeatedly challenged with aerosolized antigen solution to induce an asthmatic reaction. Twenty-four hours after the last antigen challenge, marked airway inflammation and bronchial smooth muscle hyperresponsiveness were observed. In this animal model of allergic bronchial asthma, a significant increase in the expression/activity of MMP-12 was found: the peak was observed at 12 h after the last antigen challenge. Furthermore, mRNA expression of MMP-12 was also increased at the early phase (1—3 h) after the last antigen challenge. Immunohistochemical studies revealed that MMP-12 was mainly expressed in airway epithelia and alveolar macrophages. These findings suggest that MMP-12 is upregulated after the induction of asthmatic reaction. MMP-12 might be a new target for the therapy against allergic bronchial asthma.
Notes
  • Shin Tokunaga, Yasuhiro Takeda, Tsuyoshi Niimoto, Norihisa Nishida, To ...
    2007 年 30 巻 2 号 p. 363-366
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    In the present study, we studied the effect of valerian extract preparation (BIM) containing valerian extract, golden root (Rhodiola rosea L.) extract and L-theanine (γ-glutamylethylamide) on the sleep-wake cycle using sleep-disturbed model rats in comparison with that of valerian extract. A significant shortening in sleep latency was observed with valerian extract and the BIM at a dose of 1000 mg/kg. On the other hand, valerian extract and the BIM caused no significant effects on total times of wakefulness, non-rapid eye movement (non-REM) sleep and REM sleep. Valerian extract and the BIM at a dose of 1000 mg/kg also had no significant effect on delta activity. In conclusion, it became clear that the BIM could be useful as a herbal medicine having a sleep-inducing effect without causing an alteration of the sleep-wakefulness cycle.
  • Vummidi Giridhar Premkumar, Srinivasan Yuvaraj, Kothandaraman Vijayasa ...
    2007 年 30 巻 2 号 p. 367-370
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    In breast cancer patients, it is not the primary tumour, but its metastases at distant sites that are the main cause of death. Circulating breast cancer tumour markers such as carcinoembryonic antigen (CEA) and carbohydrate antigen 15-3 (CA 15-3) are reliable indicators of impending relapse, in which an increasing tumour marker level is associated with a very likelihood of developing recurrence. In the present study, 84 breast cancer patients were randomized to receive a daily supplement of 100 mg coenzyme Q10 (CoQ10), 10 mg riboflavin and 50 mg niacin (CoRN) one dosage per day along with 10 mg tamoxifen (TAM) twice a day. Serum CEA and CA 15-3 levels were elevated in untreated breast cancer patients (group II) and their tumour marker levels significantly reduced upon tamoxifen therapy for more than 1 year (group III). Group III patients supplemented with CoRN for 45 d (group IV) and 90 d (group V) along with tamoxifen significantly reduced CEA and CA 15-3 levels. This study suggests supplementing CoRN to breast cancer patients along with tamoxifen reduces the serum tumour marker level and thereby reduce the risk of cancer recurrence and metastases.
  • Susumu Imaoka, Tomohiro Mori, Tsutomu Kinoshita
    2007 年 30 巻 2 号 p. 371-374
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Bisphenol A (BpA) is widely used in industry and dentistry. Its effects on the embryonic development of Xenopus laevis were investigated. Xenopus embryos at stage 10.5 were treated with BpA. Developmental abnormalities were observed at stage 35; malformation of the head region including eyes and scoliosis. The expression of several markers of embryonic development was investigated by reverse transcription-polymerase chain reaction (RT-PCR). The pan-neural marker SOX-2, the neural stem cell marker nrp-1, the mesodermal marker MyoD, and the endodermal marker sox17α, were used. Although the expression of marker genes was not changed by treatment with BpA, that of Pax-6, a key regulator of the morphogenesis of the eyes, was decreased by BpA. Pax-6 is a downstream factor of Notch signaling. So, the expression of a typical Notch-dependent factor, ESR-1, was investigated in the presence of BpA. The expression of ESR-1 was efficiently suppressed by BpA. In whole mount in situ hybridization (WISH), Pax-6 was expressed in the central nervous system and eyes. The expression was lost completely on treatment with BpA. The expression of ESR-1 in the central nervous system and eyes also disappeared with BpA treatment. Injection of the intracellular domain of Notch efficiently recovered ESR-1 expression in the presence of BpA although injection of a ligand for notch, Delta, did not. These results suggest that BpA decreased the expression of ESR-1 by disrupting the Notch signal.
  • Otavio Aparecido Flausino, Jr., Ana Maria Pereira, Vanderlan da Silva ...
    2007 年 30 巻 2 号 p. 375-378
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    The effects of acute oral administration of erythrinian alkaloids, i.e. (+)-α-hydroxy-erysotrine, erythravine and (+)-11α-hydroxy-erythravine isolated from the flowers of Erythrina mulungu were investigated in two animal models of anxiety in mice—the light–dark transition model (LDTM) and the elevated plus-maze (EPM). In the LDTM, erythravine (3, 10 mg/kg) and (+)-11α-hydroxy-erythravine (10 mg/kg) increased the time spent by the animals in the illuminated compartment and (+)-11α-hydroxy-erythravine (3 mg/kg) increased the number of transitions between compartments of the LDTM, suggesting an anxiolytic-like effect of these erythrinian alkaloids. Nevertheless, the third alkaloid studied, (+)-α-hydroxy-erysotrine, did not change any behavioral response with the range of doses used (3—10 mg/kg). Since the oral administration of the crude extract of E. mulungu (EM) (100—400 mg/kg) did not modify the conventional measures of anxiety in the EPM, this animal model was not chosen to evaluate the anxiolytic properties of the isolated alkaloids. These results suggest that the alkaloids erythravine and (+)-11α-hydroxy-erythravine are responsible for the anxiolytic effects of the crude extract of E. mulungu.
Regular Articles
  • Je-Min Choi, Byoung-Seok Yoon, Sang-Kyou Lee, Jae-Kwan Hwang, Ryung Ry ...
    2007 年 30 巻 2 号 p. 324-330
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Neohesperidin dihydrochalcone (NHDC), a non-nutritive sweetening agent, is simply produced by hydrogenation of neohesperidin. The aim of this study is to evaluate the antioxidant and radical scavenging properties of neohesperidin dihydrochalcone and other structurally related compounds (phloridzin, neohesperidin) toward different reactive radical and oxygen species including ·ABTS+, ·O2, ·OH, H2O2, and HOCl in vitro. NHDC showed remarkable radical scavenging activity against stable radical and reactive oxygen species (ROS) in concentration dependent manner. Especially, NHDC was the most potent inhibitor of H2O2 and HOCl. NHDC showed HOCl scavenging activity of 93.5% and H2O2 scavenging property of 73.5% which was more than those of all the tested compounds including ascorbic acid and BHT. Moreover, NHDC could inhibit protein degradation, plasmid DNA strand cleavage and HIT-T15, HUVEC cell death from HOCl attack while mannitol, BHT, and ascorbic acid could not protect them effectively. These results suggest that NHDC is a potent antioxidant, especially it is evaluated as a novel HOCl scavenger. This study implies the possibility of therapeutic effect of NHDC on ROS-related inflammatory diseases.
  • Ken Shirato, Kazuhiko Imaizumi, Akihisa Abe, Akio Tomoda
    2007 年 30 巻 2 号 p. 331-336
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    The aim of the present study was to determine whether phenoxazines such as 2-amino-4,4-α-dihydro-4α-phenoxazine-3-one (Phx-1) and 2-aminophenoxazine-3-one (Phx-3) may suppress the proliferation of human neuroblastoma cell line, NB-1 that is refractory to chemotherapeutic agents, inducing apoptosis through the activation of caspase pathway or not. Phx-1 and Phx-3 suppressed the proliferation of NB-1 cells extensively dependent on dose and time. The IC50 of Phx-1 and Phx-3 was about 20 μM and 0.5 μM, respectively, when the cells were treated with Phx-1 or Phx-3 for 72 h. Phx-1 and Phx-3 caused the mixed types of cell death—apoptosis and necrosis—in NB-1 cells, which was detected by flow cytometry. The induction of apoptosis/necrosis caused by these phenoxazines seemed to be correlated dominantly with the caspase independent pathway, because the increased activity of effector caspase 3/7 in NB-1 cells caused by 50 μM Phx-1 or 20 μM Phx-3 was completely cancelled by the addition of z-VAD-fmk, a pan-caspase inhibitor, but such phenoxazines-suppressed viability of NB-1 cells was not recovered to normal levels by this inhibitor. The results of this study demonstrate that Phx-1 and Phx-3 have antitumor activity against the neuroblastoma cell line, NB-1, though the IC50 was extremely low for Phx-3, inducing the mixed types of cell death, apoptosis and necrosis, caspase-independently.
Regular Articles
  • Young-Ju Yun, Bombi Lee, Dae-Hyun Hahm, Sung-Keel Kang, Seung-Moo Han, ...
    2007 年 30 巻 2 号 p. 337-342
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Ginseng Radix, Atractylodis Macrocephalae Rhizoma, Poria, Glycyrrhizae Radix, Angelicae Gigantis Radix, Ligusticum Rhizoma, Rehmanniae Radix, Paeoniae Radix, Acori Graminei Rhizoma, and Polygalae Radix have been widely used as herbal medicine against ischemia. In order to test the neuroprotective effect of a novel prescription, the present study examined the effects of Palmul-Chongmyeong-Tang (PMCMT) consisting of these ten herbs on learning and memory in the Morris water maze task and the central cholinergic system of rats with cerebral ischemia-induced neuronal and cognitive impairments. After middle cerebral artery occlusion (MCAO) for 2 h, rats were administered with saline or PMCMT (200 mg/kg, p.o.) daily for 2 weeks, followed by their training to the tasks. In the water maze test, the animals were trained to find a platform in a fixed position during 6 d and then received a 60 s probe trial on the 7th day following removal of the platform from the pool. Rats with ischemic insults showed impaired learning and memory of the tasks and treatment with PMCMT produced a significant improvement in escape latency to find the platform in the Morris water maze. Consistent with behavioral data, treatment with PMCMT also reduced the loss of cholinergic immunoreactivity in the hippocampus induced by cerebral ischemia. These results demonstrated that PMCMT has a protective effect against ischemia-induced neuronal and cognitive impairments. The present study suggested that PMCMT might be useful in the treatment of vascular dementia.
Notes
  • MinKyun Na, Kyung Ah Kim, Hyuncheol Oh, Bo Yeon Kim, Won Keun Oh, Jong ...
    2007 年 30 巻 2 号 p. 379-381
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Inhibition of protein tyrosine phosphatase 1B (PTP1B) has been proposed as a strategy for the treatment of type 2 diabetes and obesity. Bioassay-guided fractionation of MeOH extract of Selaginella tamariscina (Selaginellaceae) afforded a PTP1B inhibitory compound, amentoflavone. The compound inhibited PTP1B with an IC50 value of 7.3±0.5 μM. Kinetic study suggested that amentoflavone is a non-competitive inhibitor of PTP1B, with a Ki value of 5.2 μM. Treatment of 32D cells overexpressing the insulin receptor (IR) with amentoflavone resulted in a dose-dependent increase in tyrosine phosphorylation of IR. These results indicate that amentoflavone may enhance insulin-induced intracellular signaling possibly through inhibition of PTP1B activity.
  • Toru Iizuka, Masahiro Nagai, Akemi Taniguchi, Hiroyoshi Moriyama, Keik ...
    2007 年 30 巻 2 号 p. 382-384
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    A platelet-aggregatory inhibitor was isolated from the 50% MeOH extract of Phyllanthus niruri L. leaf. Its structure was determined to be methyl brevifolincarboxylate on the basis of the 1H-, 13C-NMR, and high-resolution mass spectral data. We compared the antiplatelet aggregatory effects of the constituent with adenosine, a well-known inhibitor of platelet aggregation. Platelet aggregation was induced by collagen or adenosine 5′-diphosphate as an activating agent; the extent of inhibition was monitored with a platelet aggregometer employing a laser-scattering method. The inhibitory effects of methyl brevifolincarboxylate were found to be as potent as adenosine that is known to act on an A2A subtype receptor.
  • Taiko Oda, Keiichi Kamoshita, Sakiko Maruyama, Kuniko Masuda, Masakazu ...
    2007 年 30 巻 2 号 p. 385-387
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    The in vitro growth inhibitory activity of lissoclibadins and lissoclinotoxins isolated from the tropical ascidian Lissoclinum cf. badium against nine human cancer cell lines was examined to evaluate their potential anticancer efficacy. Lissoclibadins 1 (1) and 2 (2), and lissoclinotoxin F (4) showed the strongest activity of the six compounds tested, which were more potent than the anticancer drug cisplatin. Compound 1 has a trimeric structure, and compounds 2 and 4 are structural isomers possessing dimeric structures connected by disulfide and sulfide bonds of trans- and cis-orientations, respectively. Lissoclibadin 3 (3), a dimeric compound connected by two sulfide bonds, and two monomeric compounds (5, 6) were less active than 1, 2, and 4. Lissoclibadin 2 (2) was the most interesting compound possessing potent inhibitory activity against colon (DLD-1 and HCT116), breast (MDA-MB-231), renal (ACHN), and non-small-cell lung (NCI-H460) cancer cell lines and showing no toxicity following a 50 mg/kg single treatment to mice, and preferable stability in rat plasma.
  • Qilong Wang, Hisashi Matsuda, Koudai Matsuhira, Seikou Nakamura, Dan Y ...
    2007 年 30 巻 2 号 p. 388-392
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Thunberginols A, B, and F from the processed leaves of Hydrangea macrophylla var. thunbergii (Hydrangeae Dulcis Folium) substantially inhibited the degranulations by antigen and calcium ionophore A23187, and the releases of TNF-α and IL-4 by antigen in RBL-2H3 cells. Phyllodulcin and hydrangenol also showed significant inhibition for the antigen-induced degranulations, but their effects were weaker than those of thunberginols A, B, and F. Among them, thunberginol B showed the most potent activity. With regard to structural requirements of thunberginols for the activity, the 3,4-double bond was essential for the strong activity and the 6-hydroxyl group and lactone ring enhanced the activity. Thunberginols A, B, and F inhibited increase in intracellular free Ca2+ levels, which is an essential process for the degranulation and production of cytokines, in RBL-2H3 cells induced by antigen, but not by calcium ionophore A23187. These results suggested that these active compounds inhibited the degranulation processes both before and after increase in intracellular free Ca2+ levels.
Regular Articles
  • Jia-You Fang, Tung-Hu Tsai, Yu-Ying Lin, Wen-Wen Wong, Meng-Nan Wang, ...
    2007 年 30 巻 2 号 p. 343-349
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Using in vitro and in vivo techniques, terpenes were evaluated as enhancers to improve the skin permeation of therapeutically active agents derived from tea, including tea catechins and theophylline. The in vitro permeation was determined by Franz cells. The skin deposition and subcutaneous amounts of drugs sampled by microdialysis were evaluated in vivo. Terpenes varied in their activities of enhancing drug permeation. The oxygen-containing terpenes were effective enhancers of drug permeation, whereas the hydrocarbon terpenes were much less efficient. Oxygen-containing terpenes with a bicyclic structure had reduced enhancing activity. Terpenes enhanced tea catechin permeation to a much greater degree than they did theophylline. The isomers of (+)-catechin and (−)-epicatechin showed different permeation behaviors when incorporated with terpenes. In the in vivo status, terpenes promoted the skin uptake but not the subsequent subcutaneous concentration of (−)-epigallocatechin gallate (EGCG). Both increased skin/vehicle partitioning and lipid bilayer disruption of the stratum corneum (SC) contributed the enhancing mechanisms of terpenes for topically applied tea catechins and theophylline based on the experimental results from the partition coefficient and transepidermal water loss (TEWL). α-Terpineol was found to be the best enhancer for catechins and theophylline. The high enhancement by α-terpineol was due to macroscopic perturbation of the SC and the biological reaction in viable skin as evaluated by TEWL and colorimetry.
  • Gopal Garg, Shailendra Saraf, Swarnlata Saraf
    2007 年 30 巻 2 号 p. 350-353
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Cubosomes are nanoparticles but instead of the solid particles, cubosomes are self-assembled liquid crystalline particles of certain surfactant with proper ratio of water with a microstructure that provides unique properties of practical interest. The discovery of cubosomes is a unique story and spans the field of food science, differential geometry, biological membranes and digestive processes. One of the most common surfactants used to make cubosomes is the monoglyceride glycerol monoolein. Bicontinuous cubic liquid crystalline phase is an optically clear, very viscous material that has a unique structure at the nanometer scale. The word bicontinuous refers to the division of the two continuous but non-intersecting aqueous regions by a lipid bilayer that is contorted into a space-filling structure. Hydrating a surfactant or polar lipid that forms cubic phase and then dispersing the solid-like phase into smaller particles usually form Cubosomes. There is a lot of excitement about the cubic phases because its unique microstructure is biologically compatible and capable of controlled release of solubilized active ingredients like drugs and proteins.
Notes
  • Sangkil Lee, Jaehwi Lee, Young Wook Choi
    2007 年 30 巻 2 号 p. 393-396
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    To enhance skin permeation of ascorbyl palmitate (AsP), it was encapsulated in liposomes, and formulated into liposomal hydrogel (lipogel) by dispersing the liposome into poloxamer hydrogel matrix. To improve the skin permeation of AsP, we applied electric current supplying system that mimics an electric skin massager. We evaluated the effects of composition and surface charge of the liposomes and electrical assistance on the skin permeation of AsP. In the passive transport study, the permeated amounts of AsP from all the lipogels tested were higher than that of control hydrogel which contains Transcutol® used to solubilize AsP. In the cathodal delivery condition with a fixed cathodal current of 0.4 mA/cm2, the skin permeation characteristics of the negative lipogels were superior to that obtained with the neutral lipogels and the drug permeation was more increased with increased surface negative charge of the liposomes. In conclusion, the lipogel system was thought as a helpful drug delivery system to enhance skin permeation of AsP. Combined use of negative lipogel with cathodal electric assistance was found to be promising in enhancing the skin delivery of AsP.
  • Zeng-Chun Ma, Hong-Ling Tan, Cheng-Rong Xiao, Yu-Guang Wang, Sheng-Qi ...
    2007 年 30 巻 2 号 p. 397-402
    発行日: 2007年
    公開日: 2007/02/01
    ジャーナル フリー
    Tetramethylpyrazine is the active ingredient of a Chinese herbal medicine. In this study, tetramethylpyrazine was tested for its activities in irradiated bone marrow stromal QXMSC1 cells. The proliferation of QXMSC1 cells was measured by MTS assay kit and flow cytometry. To identify proteins involved in the processes of cellular and molecular response of tetramethylpyrazine to irradiation damage, we comparatively analyzed the proteome of nonirradiated, irradiated and tetramethylpyrazine treated QXMSC1 cells. Reverse transcriptase polymerase chain reaction (RT-PCR) were used to validate the differentially expressed proteins. 20 Gy 60Co γ irradition inhibited QMSC1 cells growth and tetramethylpyrazine could reverse of this action due to stimulating QXMSC1 cells from G1 to S progression. Proteomic analytical results showed that 18 spots were changed in irradiated QXMSC1 cells, and 15 spots matched with known proteins after database searching. The expression level of proteins such as translationally controlled tumor protein (TCTP), and galectin-3, were increased in irradiated QXMSC1 cells, while calmodulin, pyruvate kinase were decreased. Tetramethylpyrazine could prevent this change or reverse to some degree. The function of these proteins involves in hematopoiesis, cell cycle and signal transduction. The changes of these proteins were confirmed by RT-PCR at mRNA levels. This study suggested that stimulating proliferation via tetramethylpyrazine played an important role in the cure effect on irradiated QXMSC1 cells and was helpful to deeply understand the mechanism of tetramethylpyrazine at the molecular level.
feedback
Top