研究者詳細 - 沢　禎彦

論文 - 沢　禎彦

分割表示 95 件中 61 - 80 件目／全件表示 >>

EMILIN-1 regulates the amount of oxytalan fiber formation in periodontal ligaments in vitro 査読国際誌

Yuka Nakatomi, Eichi Tsuruga, Kazuki Nakashima, Yoshihiko Sawa, Hiroyuki Ishikawa

Connective Tissue Research 52 ( 1 ) 30 - 35 2011年2月

記述言語：英語掲載種別：研究論文（学術雑誌）

The elastic system fibers comprise oxytalan, elaunin, and elastic fibers, differing in their relative microfibril and elastin contents. Among them, human periodontal ligament (PDL) contains only oxytalan fibers (pure microfibrils). Elastin microfibril interface-located protein-1 (EMILIN-1) is localized at the interface between microfibrils and elastin. We hypothesized that EMILIN-1 may contribute to the formation of oxytalan fibers. We used a small interfering RNA (siRNA) for EMILIN-1 in PDL cell culture to examine the extracellular deposition of fibrillin-1 (the major component of microfibrils). EMILIN-1 was labeled on microfibrils positive for fibrillin-1 and was colocalized with fibrillin-1 upon immunoprecipitation assay. EMILIN-1 suppression reduced the level of fibrillin-1 deposition to 23 of the control, and this was responsible for the diminution of fibrillin-1 deposition revealed by immunofluorescence. These results suggest that EMILIN-1 may regulate the formation of oxytalan fibers and play a role in their homeostasis. © 2011 Informa Healthcare USA, Inc.

DOI： 10.3109/03008207.2010.502982

Expression of podoplanin and classical cadherins in salivary gland epithelial cells of klotho-deficient mice 査読

Ikuko Amano, Yuri Imaizumi, Chiaki Kaji, Hiroshi Kojima, Yoshihiko Sawa

Acta Histochemica et Cytochemica 44 ( 6 ) 267 - 276 2011年

記述言語：英語掲載種別：研究論文（学術雑誌）

We have recently shown that salivary gland myoepithelial cells express podoplanin. Podoplanin indirectly binds the actin filament network which links classical cadherins. The study here is aimed to investigate the expression of podoplanin and cadherins on salivary gland myoepithelial cells and the changes in the aging cells using klotho-deficient (kl/kl) mice. The submandibular glands of kl/kl mouse lack granular ducts which express klotho in wild type mice, suggesting that klotho may be a gene responsible for granular duct development. Although aging resulted in growth suppression of myoepithelial cells because of the sparse distribution of the cells in kl/kl mouse salivary glands, the expression of podoplanin and Ecadherin was shown in aging myoepithelial cells. It is thought that podoplanin participates in the actin-E-cadherin networks which are maintained in aging myoepithelial cells. It was also shown that granular ducts were filled with P-cadherin, and that the P-cadherin amount was larger in the wild type mouse submandibular glands than in the sublingual and parotid glands of wild type mouse, and in the submandibular glands of kl/kl mouse. These findings suggest that the granular duct is an organ secreting soluble P-cadherin into the saliva. © 2011 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.11037

The effect of valproic Acid on mesenchymal pluripotent cell proliferation and differentiation in extracellular matrices. 国際誌

Yuji Hatakeyama, Junko Hatakeyama, Atsushi Takahashi, Kyoko Oka, Eichi Tsuruga, Tetsuichiro Inai, Yoshihiko Sawa

Drug target insights 5 1 - 9 2011年

記述言語：英語掲載種別：研究論文（学術雑誌）

Valproic acid (2-n-propylpentanoic acid, VPA) is a widely used antiepileptic and anticonvulsant drug. Previous studies have reported that VPA effects osteogenesis in vivo and in vitro, yet it remains unclear whether VPA promotes cell differentiation of osteoblasts derived from mesenchymal cells. The purpose of this study was to clarify the effect of VPA on undifferentiated pluripotent mesenchymal cell proliferation and differentiation into osteoblasts while analyzing the impact of the absence or presence of extracellular matrices (ECMs). Mouse mesenchymal cells were cultured on non-coated plastic, type I collagen-coated, and fibronectin-coated plates in the absence or presence of VPA. A cell proliferation assay was performed in which modified formazan dye content was analyzed and proliferation nuclear antigen (PCNA)-positive cells were counted at various concentrations of VPA. A high concentration of VPA did not clearly alter cell morphology, but large numbers of stress fibers were observed in these cells and the cell proliferation ratio was decreased with positive PCNA counts. In the presence of matrices, the cell proliferation ratio decreased at low VPA concentrations compared with the ratio obtained in the absence of these ECMs. On the other hand, VPA promoted osteoblastic differentiation in the presence of type I collagen. These findings indicate that for undifferentiated mesenchymal cells, VPA promotes a decrease in the cell proliferation rate in the presence of ECMs and promotes osteoblastic differentiation, both of which could provide insight into additional mechanisms of osteoblastic cell differentiation caused by VPA.

DOI： 10.4137/DTI.S6534

歯根膜オキシタラン線維形成過程におけるEMILIN-1の機能

中冨佑香, 中島一記, 久永豊, 敦賀英知, 沢禎彦, 石川博之

日本矯正歯科学会大会プログラム・抄録集 69回 184 - 184 2010年9月

Immunoelectron microscopic study of podoplanin localization in mouse salivary gland myoepithelium 査読

Minoru Hata, Ikuko Amano, Eichi Tsuruga, Hiroshi Kojima, Yoshihiko Sawa

Acta Histochemica et Cytochemica 43 ( 2 ) 77 - 82 2010年

記述言語：英語掲載種別：研究論文（学術雑誌）

We have recently reported that salivary gland cells express the lymphatic endothelial cell marker podoplanin. The present study was aimed to immunohistochemically investigate the expression of the myoepithelial cell marker α-smooth muscle actin (SMA) on podoplaninpositive cells in mouse parotid and sublingual glands, and to elucidate podoplanin localization in salivary gland myoepithelial cells by immunoelectron microscopic study. The distribution of myoepithelial cells expressing podoplanin and α-SMA was examined by immunofluorescent staining, and the localization of reaction products of anti-podoplanin antibody was investigated by pre-embedded immunoelectron microscopic method. In immunohistochemistry, the surfaces of both the mucous acini terminal portion and ducts were covered by a number of extensive myoepithelial cellular processes expressing podoplanin, and the immunostaining level with anti-podoplanin antibody to myoepithelial cells completely coincided with the immunostaining level with anti-α-SMA antibody. These findings suggest that podoplanin is a salivary gland myoepithelial cell antigen, and that the detection level directly reflects the myoepithelial cell distribution. In immunoelectron microscopic study, a number of reaction products with anti-podoplanin antibody were found at the Golgi apparatus binding to the endoplasmic reticulum in the cytoplasm of myoepithelial cells between sublingual gland acinar cells, and were also found at the myoepithelial cell membrane. These findings suggest that salivary gland myoepithelial cells constantly produce podoplanin and glycosylate at the Golgi apparatus, and transport them to the cell membrane. Podoplanin may be involved in maintaining the homeostasis of myoepithelial cells through its characteristic as a mucin-type transmembrane glycoprotein. © 2010 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.10011

Fibulin-4 and -5, but not Fibulin-2, are Associated with Tropoelastin Deposition in Elastin-Producing Cell Culture 査読

Yoshinori Yamauchi, Eichi Tsuruga, Kazuki Nakashima, Yoshihiko Sawa, Hiroyuki Ishikawa

Acta Histochemica et Cytochemica 43 ( 6 ) 131 - 138 2010年

記述言語：英語掲載種別：研究論文（学術雑誌）

201E0l aTshteic J sapyasnte Smo cfiibeteyr so fc oHnisstioscth oefm misitcrryo afinbdri lCs ya-nd tropoelastin. During development, microfibrils act as a template on which tropoelastin is deposited. Fibrillin-1 is the major component of microfibrils. It is not clear whether elastic fiber-associated molecules, such as fibulins, contribute to tropoelastin deposition. Among the fibulin family, fibulin-2, -4 and -5 are capable of binding to tropoelastin and fibrillin-1. In the present study, we used the RNA interference (RNAi) technique to establish individual gene-specific knockdown of fibulin-2, -4 and -5 in elastin-producing cells (human gingival fibroblasts
HGF). We then examined the extracellular deposition of tropoelastin using immunofluorescence. RNAi-mediated downregulation of fibulin-4 and -5 was responsible for the diminution of tropoelastin deposition. Suppression of fibulin-5 appeared to inhibit the formation of fibrillin-1 microfibrils, while that of fibulin-4 did not. Similar results to those for HGF were obtained with human dermal fibroblasts. These results suggest that fibulin-4 and -5 may be associated in different ways with the extracellular deposition of tropoelastin during elastic fiber formation in elastinproducing cells in culture. © 2010 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.10026

Immunohistochemical examination on the distribution of cells expressed lymphatic endothelial marker podoplanin and LYVE-1 in the mouse tongue tissue 査読

Yuya Noda, Ikuko Amano, Minoru Hata, Hiroshi Kojima, Yoshihiko Sawa

Acta Histochemica et Cytochemica 43 ( 2 ) 61 - 68 2010年

記述言語：英語掲載種別：研究論文（学術雑誌）

The clinical study for lingual disease requires the detailed investigation of the lingual lymphatic network and lymphatic marker-positive cells. Recently, it has been reported that several tissue cells and leukocytes express lymphatic markers, LYVE-1 and podoplanin. This study was aimed to clarify the lingual distribution of cells expressing LYVE-1 and podoplanin. In the mouse tongue, podoplanin is expressed in nerve sheaths, lingual gland myoepithelial cells, and lymphatic vessels. LYVE-1 is expressed in the macrophage marker Mac-1-positive cells as well as lymphatic vessels, while factor-VIII was detected in only blood endothelial cells. α-SMA was detected in vascular smooth muscle and myoepithelial cells. Therefore, identification of lymphatic vessels in lingual glands, the combination of LYVE-1 and factor-VIII, or LYVE-1 and Mac-1 is useful because myoepithelial cells express podoplanin and α-SMA. The immunostaining of factor-VIII on lymphatic vessels was masked by the immunostaining to LYVE-1 or podoplanin because lymphatic vessels express factor-VIII to a far lesser extent than blood vessels. Therefore, except for the salivary glands, the combination of podoplanin and α-SMA, or factor-VIII is useful to identify lymphatic vessels and blood vessels with smooth muscle, or blood capillaries. © 2010 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.10008

Immunohistochemical examination for the distribution of podoplanin-expressing cells in developing mouse molar tooth germs 査読

Yuri Imaizumi, Ikuko Amano, Eichi Tsuruga, Hiroshi Kojima, Yoshihiko Sawa

Acta Histochemica et Cytochemica 43 ( 5 ) 115 - 121 2010年

記述言語：英語掲載種別：研究論文（学術雑誌）

We recently reported the expression of podoplanin in the apical bud of adult mouse incisal tooth. This study was aimed to investigate the distribution of podoplanin-expressing cells in mouse tooth germs at several developing stages. At the bud stage podoplanin was expressed in oral mucous epithelia and in a tooth bud. At the cap stage podoplanin was expressed on inner and outer enamel epithelia but not in mesenchymal cells expressing the neural crest stem cell marker nestin. At the early bell stage nestin and podoplanin were expressed in cervical loop and odontoblasts. At the root formation stage both nestin and podoplanin were weakly expressed in odontoblasts generating radicular dentin. Podoplanin expression was also found in the Hertwig epithelial sheath. These results suggest that epithelial cells of developing tooth germ acquire the ability to express nestin, and that tooth germ epithelial cells maintain the ability to express podoplanin in oral mucous epithelia. The expression of podoplanin in odontoblasts was induced as tooth germ development advanced, but was suppressed with the completion of the primary dentin, suggesting that podoplanin may be involved in the cell growth of odontoblasts. Nestin may function as an intermediate filament that binds podoplanin in odontoblasts. © 2010 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.10023

Fibulin-5 contributes to microfibril assembly in human periodontal ligament cells 査読

Yutaka Hisanaga, Kazuki Nakashima, Eichi Tsuruga, Yuka Nakatomi, Yuji Hatakeyama, Hiroyuki Ishikawa, Yoshihiko Sawa

Acta Histochemica et Cytochemica 42 ( 5 ) 151 - 157 2009年10月

記述言語：英語掲載種別：研究論文（学術雑誌）

The elastic system fibers comprise oxytalan, elaunin and elastic fibers, which differ in their relative microfibril and elastin content. Human periodontal ligaments (PDL) contain only oxytalan fibers (pure microfibrils) among them. Since fibulin-5 regulates the organization of elastic fibers to link the fibers to cells, we hypothesized that fibulin-5 may contribute to the formation of oxytalan fibers. We used siRNA for fibulin-5 in PDL cell culture to examine the extracellular deposition of fibrillin-1 and -2, which are the major components of microfibrils. Fibulin-5 was labeled on microfibrils positive for fibrillin-1 and -2. Fibulin-5 suppression reduced the level of fibrillin-1 and -2 deposition to 60% of the control level. These results suggest that fibulin-5 may control the formation of oxytalan fibers, and play a role in the homeostasis of oxytalan fibers. © 2009 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.09021

Stretching stimulates fibulin-5 expression and controls microfibril bundles in human periodontal ligament cells 査読

K. Nakashima, E. Tsuruga, Y. Hisanaga, H. Ishikawa, Y. Sawa

Journal of Periodontal Research 44 ( 5 ) 622 - 627 2009年10月

記述言語：英語掲載種別：研究論文（学術雑誌）

Background and Objective: The elastic fiber system comprises oxytalan, elaunin and elastic fibers, differing in their relative microfibril and elastin contents. Human periodontal ligaments contain oxytalan fibers (pure microfibrils). Periodontal ligaments are continuously exposed to various functional forces, such as tooth movement and occlusal loading. We have reported that bundles of microfibrils coalesce in response to mechanical strain in cultured periodontal ligament fibroblasts, as assessed in terms of their positivity for fibrillin-1 (the major component of microfibrils). However, the mechanism of microfibril coalescence is unclear. We hypothesized that the fibrillin-1-binding molecule, fibulin-5, contributes to oxytalan fiber formation under mechanical stra Material and Methods: We subjected periodontal ligament fibroblasts to stretching in order to examine the effects of fibulin-5 on the formation of oxytalan fibers in cell/matrix layers. We transfected periodontal ligament cells with small interference RNA for fibulin-5, then examined oxytalan fibers using immunofluorescence and electron microscopy. Results: Immunofluorescence showed that fibrillin-1-positive microfibrils coalesced as a result of stretching, compared with cells that were not subjected to stretching. Fibulin-5 colocalized on fibrillin-1-positive microfibrils. Stretching increased fibulin-5 gene expression and protein deposition. Immunofluorescence and immunogold electron microscopy analysis revealed that fibulin-5 suppression inhibited the coalescence of microfibrils under stretching conditions. Conclusion: These results suggest that fibulin-5 up-regulated in response to tension strain may control the formation of microfibril bundles in periodontal ligament. © 2008 Blackwell Munksgaard.

DOI： 10.1111/j.1600-0765.2008.01170.x

Integrin αvβ3 regulates microfibril assembly in human periodontal ligament cells 査読

E. Tsuruga, A. Sato, T. Ueki, K. Nakashima, Y. Nakatomi, H. Ishikawa, T. Yajima, Y. Sawa

Tissue and Cell 41 ( 2 ) 85 - 89 2009年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier Ltd

Fibrillin-1 is the major structural component of extracellular microfibrils. However, the mechanism by which extracellular fibrillin-1 assembles into microfibrils is not fully understood. Fibrillin-1 contains the Arg-Gly-Asp (RGD) motif, which may allow binding to RGD-recognizing integrins. We hypothesized that integrin αvβ3 on the cell surface of human periodontal ligament (PDL) fibroblasts may influence fibrillin-1 assembly into cell/matrix layers. We treated PDL fibroblasts with an integrin αvβ3-specific antagonist to examine fibrillin-1 assembly. Western blotting and immunofluorescence analysis showed that treatment with the integrin αvβ3 antagonist at 5 μM clearly abolished fibrillin-1 deposition. These results provide for the first time evidence that integrin αvβ3 regulates extracellular assembly of fibrillin-1, thereby modulating cell-mediated homeostasis of microfibrils. © 2008 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.tice.2008.07.005

Expression of podoplanin in the mouse tooth germ and apical bud cells 査読

Yoshihiko Sawa, Kana Iwasawa, Hiroyuki Ishikawa

Acta Histochemica et Cytochemica 41 ( 5 ) 121 - 126 2008年10月

記述言語：英語掲載種別：研究論文（学術雑誌）

This study was designed to investigate the distribution of cells expressing podoplanin in the mouse tooth bud. Podoplanin expression was detected in enamel epithelia of the cervical loop at cell-cell contacts strongly, and weakly on the loosely aggregated stellate reticulum in the center and the neighboring stratum intermedium. Odontoblasts exhibited intense podoplanin expression at the junction with predentin while no expression was detected in the enamel organ containing ameloblasts. These results suggest that proliferating inner and outer enamel epithelia express podoplanin but that the expression is suppressed in the differentiated epithelia containing ameloblasts. On the other hand the podoplanin expression occurs in the differentiating odontoblasts and the expression is sustained in differentiated odontoblasts, indicating that odontoblasts have the strong ability to express podoplanin. In cultured apical bud cells podoplanin was detected at cell-cell contacts. In real-time PCR analysis the amount of podoplanin mRNA of the apical buds was 2-fold compared with the amount of kidney used as a positive control. These findings indicate that apical bud cells have the strong ability to express the podoplanin gene. Podoplanin is a mucin-type glycoprotein negatively charged by extensive O-glycosylation and a high content of sialic acid, which expresses the adhesive property. The podoplanin may contribute to form odontoblastic fiber or function as the anchorage to the tooth development and in proliferating epithelial cells of cervical loop and apical bud. © 2008 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.08019

Induction of ICAM-1 and VCAM-1 on the mouse lingual lymphatic endothelium with TNF-α 査読

Kana Iwasawa, Takeshi Kameyama, Hiroyuki Ishikawa, Yoshihiko Sawa

Acta Histochemica et Cytochemica 41 ( 5 ) 115 - 120 2008年10月

記述言語：英語掲載種別：研究論文（学術雑誌）

This study investigated the TNF-α-induced ICAM-1 and VCAM-1 expression on mouse lingual lymphatic vessels. All podoplanin-positive lymphatic vessels expressed PECAM-1. In the lamina propria mucosae of TNF-α-treated tongue, almost all initial lymphatics expressed ICAM-1. There were initial lymphatics with the VCAM-1 expression and also the vessels without the expression. In the tunica muscularis of TNF-α-treated tongue, collecting lymphatic vessels expressed ICAM-1, but rarely expressed VCAM-1 whereas blood vessels simultaneously expressed ICAM-1 and VCAM-1. The ICAM-1-positive rate increased with TNF-α to 75% from 10% on initial lymphatics, and to 40% from 0% on collecting lymphatic vessels while it increased to 90% from 45% on blood vessels. The VCAM-1-positive rate increased with TNF-α to 30% from 0% on initial lymphatics, and to 5% from 0% on collecting lymphatic vessels while it increased to 75% from 5% on blood vessels. These findings suggest that the lingual lymphatic endothelium has the ability to express ICAM-1, and VCAM-1 to a lesser extent than the ICAM-1 induction with TNF-α, and that the ICAM-1 and VCAM-1 induction predominantly occurs in the initial lymphatics compared with collecting lymphatic vessels. © 2008 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.08017

Expression of podoplanin in the mouse salivary glands 査読国際誌

Minoru Hata, Takeshi Ueki, Atsuko Sato, Hiroshi Kojima, Yoshihiko Sawa

Archives of Oral Biology 53 ( 9 ) 835 - 841 2008年9月

記述言語：英語掲載種別：研究論文（学術雑誌）

Objective: Podoplanin is one of the most highly expressed lymphatic-specific genes. Here, we report the distribution of cells expressing podoplanin in mouse salivary glands. Design: We immunohistochemically investigated the distribution of cells expressing podoplanin in mouse major salivary glands by laser-scanning microscopy. The expression of endothelial cell marker PECAM-1 was tested to discriminate lymphatic endothelium from salivary gland cells, and myoepithelial cells were identified by an antibody for P-cadherin. Results: The podoplanin expression was rarely found in acini of the parotid gland but clearly found at the basal portion of acini in the submandibular and sublingual glands. The number of portion reacted with anti-podoplanin is greater in the sublingual gland than in the submandibular gland. The expression was also found at the basal portion of ducts in all major salivary glands. The P-cadherin expression was rarely found in acini of the parotid gland but found in acini of the sublingual gland and on ducts in parotid and sublingual glands, corresponding to the area of podoplanin expression. Conclusions: It was suggested that the acinar and myoepithelial cells in the salivary glands have the ability to express podoplanin, and that the expression may be concerned with the mucous saliva excretion. © 2008 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.archoralbio.2008.02.006

Leukocyte adhesion molecule and chemokine production through lipoteichoic acid recognition by toll-like receptor 2 in cultured human lymphatic endothelium 査読国際誌

Yoshihiko Sawa, Eichi Tsuruga, Kana Iwasawa, Hiroyuki Ishikawa, Shigemitsu Yoshida

Cell and Tissue Research 333 ( 2 ) 237 - 252 2008年8月

記述言語：英語掲載種別：研究論文（学術雑誌）

We have recently reported that the human lymphatic endothelium has toll-like receptor 4 (TLR4)-mediated lipopolysaccharide recognition mechanisms that induce the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Although ligand engagement with TLR2 enables activation of the MyD88-dependent pathway similarly to TLR4, whether TLR2 ligands such as lipoteichoic acid (LTA) trigger the activation of lymphatic endothelium remains unclear. This study has been designed to investigate the expression dynamics of LTA-induced leukocyte adhesion molecules and chemokines in cultured human lymphatic endothelium (LEC). Reverse transcription/polymerase chain reaction (RT-PCR) and real-time quantitative PCR analyses have shown that LEC usually expresses TLR2 and increases TLR2 gene expression on LTA treatment. Indeed, LTA-treated LEC increases the expression of E-selectin, ICAM-1, and VCAM-1 but does not alter the gene expression of ICAM-2, ICAM-3, junctional adhesion molecule-1 (JAM-1), JAM-3, or platelet endothelial cell adhesion molecule-1 (PECAM-1). The expression of LTA-induced E-selectin, ICAM-1, and VCAM-1 in LEC is suppressed by anti-TLR2 but not by anti-TLR4 and is also suppressed by TLR2-specific short interfering RNA (siRNA) but not by siRNA for TLR4. The expression of CCL2, CCL5, and CCL20 (Cys-Cys motif chemokines) and of CXCL1, CXCL3, CXCL5, CXCL6, and CXCL8 (Cys-X-Cys motif chemokines) was induced in LEC with LTA. These data suggest that the human lymphatic endothelial phenotype has TLR2-mediated LTA-recognition mechanisms, resulting in increased expression of inflammatory leukocyte adhesion molecules and phagocyte-attractive chemokines. The human lymphatic endothelium may thus function to collect leukocytes from tissues into lymphatic vessels by means of immunologically functional molecules. © 2008 Springer-Verlag.

DOI： 10.1007/s00441-008-0625-5

The expression of E-selectin and chemokines in the cultured human lymphatic endothelium with lipopolysaccharides 査読国際誌

Yoshihiko Sawa, Eichi Tsuruga

Journal of Anatomy 212 ( 5 ) 654 - 663 2008年5月

記述言語：英語掲載種別：研究論文（学術雑誌）

This study investigated the expression of selectins and chemokines in cultured human lymphatic endothelial cells stimulated with lipopolysaccharides. In microarray, vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 gene expressions in the lymphatic endothelium with lipopolysaccharides did not change at 0.5 h but increased two- to three-fold at 12 h, whereas E-selectin increased 10-fold at 0.5 h and 68-fold at 12 h compared with untreated cells. The E-selectin mRNA and protein increased in the lymphatic endothelial cells with lipopolysaccharides at more than two-fold levels compared with human umbilical vein endothelial cells. Induction of Cys-Cys chemokine ligand 2, 3, 5, 7, 8 and 20 mRNAs in the lymphatic endothelial cells with lipopolysaccharides was detected in microarray and real-time PCR. The Cys-Cys chemokine ligand 2, 5 and 20 mRNA amounts in cells with high concentration lipopolysaccharides were larger in the lymphatic endothelial cells than in human umbilical vein endothelial cells. The Cys-Cys chemokine ligand 3 and 8 mRNAs were not detected in human umbilical vein endothelial cells. Induction of Cys-X-Cys chemokine ligand 1, 3, 5, 6 and 8 mRNAs was detected in the lymphatic endothelial cells with lipopolysaccharides. The Cys-X-Cys chemokine ligand 3, 5 and 8 mRNA amounts in cells with high concentration lipopolysaccharides were larger in the lymphatic endothelial cells than in human umbilical vein endothelial cells. In conclusion, it was demonstrated that the cultured human lymphatic endothelial cells express E-selectin and phagocyte-attractive chemokine genes. © 2008 The Authors Journal compilation © 2008 Anatomical Society of Great Britain and Ireland.

DOI： 10.1111/j.1469-7580.2008.00892.x

Expression of junctional adhesion molecules on the human lymphatic endothelium 査読国際誌

Takeshi Ueki, Kana Iwasawa, Hiroyuki Ishikawa, Yoshihiko Sawa

MICROVASCULAR RESEARCH 75 ( 2 ) 269 - 278 2008年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE

Human lymphatic vessels express several leukocyte adhesion molecules. The study here investigated the expression of three junctional adhesion molecules (JAM) which are a newly reported glycoprotein family of adhesion molecules on human lymphatic endothelium. In this study, JAM-1 and JAM-3 but not JAM-2 were detected in cultured human neonatal dermal lymphatic endothelial cells (LEC) at the gene and protein levels by microarray, RT-PCR, real-time PCR, and immunohistochemical analysis. The JAM-I and JAM-3 expression was not altered in the TNF-alpha-treated LEC or in the untreated cells. In human tissue, the expression of JAM-1, and the expression of JAM-1, JAM-2, and JAM-3 were observed in collecting lymphatic vessels of uninflamed small intestine, and in initial lymphatics of inflamed tongue and uninflamed gingival tissue. It is thought that JAM-2 mRNA could be produced in mature vascular endothelium but not in cultured cells, and that human intestinal and oral lymphatic vessels usually express JAM-1, JAM-2, and JAM-3. There were initial lymphatics simultaneously expressing JAM-1, JAM-2, and JAM-3 in the mucosal connective tissue papillae of gingival tissue. The three JAM expressions on the lymphatic endothelium may contribute to both seal the cell-cell contact at interendothelial junctions and also allow lymphocytes to transmigrate into lymphatic vessels from tissue, independent of inflammatory cytokines. (c) 2007 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.mvr.2007.07.005

LPS-induced IL-6, IL-8, VCAM-1, and ICAM-1 expression in human lymphatic endothelium 査読国際誌

Yoshihiko Sawa, Takeshi Ueki, Minoru Hata, Kana Iwasawa, Eichi Tsuruga, Hiroshi Kojima, Hiroyuki Ishikawa, Shigemitsu Yoshida

Journal of Histochemistry and Cytochemistry 56 ( 2 ) 97 - 109 2008年2月

記述言語：英語掲載種別：研究論文（学術雑誌）

We have previously reported the TLR4 expression in human intestinal lymphatic vessels. In the study here, microarray analysis showed the expression of the TLR4, MD-2, CD14, MyD88, TIRAP, TRAM, IRAK1, and TRAF6 genes in cultured human neonatal dermal lymphatic microvascular endothelial cells (LEC). The microarray analysis also showed that LEC expressed genes of IL-6, IL-8, VCAM-1, and ICAM-1, and the real-time quantitative PCR analysis showed that mRNA production was increased by lipopolysaccharide (LPS). The LPS-induced IL-6, IL-8, VCAM-1, and ICAM-1 production in LEC was suppressed by the introduction of TLR4-specific small interfering RNA, and also by anti-TLR4, nobiletin, and CAPE pretreatment. These findings suggest that LEC has TLR4-mediated LPS recognition mechanisms that involve at least activation of NF-κB, resulting in increased expression of IL-6, IL-8, VCAM-1, and ICAM-1. Both the LPS effect on the gene expression and also the suppression by nobiletin and CAPE pre-treatment on the protein production were larger in IL-6 and in VCAM-1 than in IL-8 and in ICAM-1 in LEC. The signal transduction of NF-κB and AP-1-dependent pathway may be more critical for the expression of IL-6 and VCAM-1 than that of IL-8 and ICAM-1 in LEC. © The Histochemical Society, Inc.

DOI： 10.1369/jhc.7A7299.2007

Effects of bFGF on suppression of collagen type I accumulation and scar tissue formation during wound healing after mucoperiosteal denudation of rat palate 査読国際誌

Wookjin Choi, Hitoshi Kawanabe, Yoshihiko Sawa, Kunihisa Taniguchi, Hiroyuki Ishikawa

ACTA ODONTOLOGICA SCANDINAVICA 66 ( 1 ) 31 - 37 2008年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：TAYLOR & FRANCIS AS

Objective. The purpose of this study was to evaluate the effect of basic fibroblast growth factor (bFGF) on collagen changes after mucoperiosteal denudation of rat palate. Material and methods. A total of 36 male Wistar rats were divided into control, scar, sham, and bFGF groups. In the scar, sham, and bFGF groups, lateral palatal mucoperiosteum was excised to form scar tissue on the palate. In the bFGF group, bFGF solution was injected into the operated area 1 week postoperatively. At 6 weeks postoperatively, the distribution of collagen type I and the 3-dimensional structure of collagen fibers were investigated under immunofluorescent and scanning electron microscopy. Result. In the bFGF group, weakly immunostained submucosa was clearly distinguishable from the strongly immunostained cervical periodontal ligament and gingiva. Collagen fibers running from submucosal tissue into the surface of underlying palatal bone comprised loosely arranged collagen fibrils. Lumen structures in collagen fibers resembled those in the control group. Conclusion. Administration of bFGF for suppression of collagen type I generation could suppress scar tissue formation and reduce connective strength with adjacent teeth and palatal bone.

DOI： 10.1080/00016350701884612

Effects of TNF-alpha on leukocyte adhesion molecule expressions in cultured human lymphatic endothelium 査読国際誌

Yoshihiko Sawa, Yukitaka Sugimoto, Takeshi Ueki, Hiroyuki Ishikawa, Atuko Sato, Toshikazu Nagato, Shigemitsu Yoshida

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY 55 ( 7 ) 721 - 733 2007年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：HISTOCHEMICAL SOC INC

TNF-alpha alters leukocyte adhesion molecule expression of cultured endothelial cells like human umbilical vein endothelial cells (HUVEC). This study was designed to investigate the changes in vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and platelet endothelial cell adhesion molecule-1 (PECAM-1) expression with TNF-alpha stimulation in cultured human neonatal dermal lymphatic endothelial cells (HNDLEC). The real-time quantitative PCR analysis on HNDLEC showed that TNF-alpha. treatment leads to increases of VCAM-1 and ICAM-1 mRNAs to the 10.8- and 48.2-fold levels of untreated cells and leads to a reduction of PECAM-1 mRNA to the 0.42-fold level of untreated cells. Western blot and immunohistochemical analysis showed that TNF-a leads to VCAM-1 and ICAM-1 expressions that were inhibited by antiserum to human TNF receptor or by AP-1 inhibitor nobiletin. In flow cytometry analysis, the number of VCAM-1- and ICAM-1-positive cells increased, and PECAM-1-positive cells decreased with TNF-alpha treatment. Regarding protein amounts produced in cells and amounts expressed on the cell surface, VCAM-1 and ICAM-1 increased in HNDLEC and HUVEC, and PECAM-1 decreased in HNDLEC in a TNF-a concentration-dependent manner. VCAM-1, ICAM-1, and PECAM-1 protein amounts in TNF-alpha-stimulated cells were lower in HNDLEC than in HUVEC. This suggests that the lymphatic endothelium has the TNF-alpha-induced signaling pathway, resulting in increased VCAM-1 and ICAM-1 expression to a weaker extent than blood endothelium and PECAM-1 reduction to a stronger extent than blood endothelium.

DOI： 10.1369/jhc.6A7171.2007

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