Biomedical Research
Online ISSN : 1880-313X
Print ISSN : 0388-6107
ISSN-L : 0388-6107
26 巻 , 6 号
December
選択された号の論文の6件中1~6を表示しています
Full Papers
  • Yoichiro KAMIYA, Kazumasa OHTA, Yuzuru KANEKO
    2005 年 26 巻 6 号 p. 231-239
    発行日: 2005年
    公開日: 2006/01/16
    ジャーナル フリー
    Local anesthetics are known to affect a variety of cellular responses other than the action of anesthetics through the Na+ channel blockade. In this study, we examined the effect of a common local anesthetic lidocaine on the cellular activity and viability of human histiocytic lymphoma U937 cells. The cellular activity and viability were assessed by WST-1 reduction activity and trypan blue exclusion test, respectively. Induction of apoptosis was monitored by DNA ladder formation, reduction of mitochondrial transmembrane potential (ΔΨm), caspase-3 activity and nuclear morphology. Lidocaine at concentrations below 12 mM induced apoptosis characterized by DNA fragmentation and chromatin condensation dose- and time-dependently. A pan-caspase inhibitor and a caspase-3 inhibitor blocked DNA ladder formation followed by the reduction of cell death. However, the caspase inhibitors did not affect the ΔΨm, but cyclosporin A inhibited the collapse of ΔΨm followed by a reduction of cell death. Lidocaine-induced apoptosis was mitochondria- and caspase-dependent, but the collapse of ΔΨm was independent of caspase activation. At concentrations above 15 mM, lidocaine induced necrosis with early disruption of membrane integrity. These results indicate that lidocaine induced apoptosis and necrosis in U937 cells depending on its dosage.
  • Shusuke KODA, Shinhiro TAKEDA, Hiroshi ONIMARU, Shinji AKADA, Atsuhiro ...
    2005 年 26 巻 6 号 p. 241-247
    発行日: 2005年
    公開日: 2006/01/16
    ジャーナル フリー
    Previous studies have suggested that cannabinoid compounds are anticonvulsants and that these compounds depress respiratory activity. However, the anticonvulsant potential of cannabinoids and their depressive effect on respiration have not been evaluated simultaneously. In the present study, we used a brainstem-spinal cord preparation model to investigate changes in inspiratory activity and the anticonvulsant effects of a cannabinoid receptor agonist, WIN55, 212-2, in bicuculline-induced convulsion. Application of 10 μM WIN55, 212-2 caused no change in inspiratory activity (6.9 ± 0.89 bursts/min vs. 8.0 ± 1.3 bursts/min, not significant) and decreased bicuculline-induced seizure-like nerve activity (number of seizure-like activities in 10 min, 11 ± 7.4 bursts vs. 1.5 ± 1.6 bursts, P < 0.01; average duration of seizure-like activity, 8.9 ± 4.0 sec vs. 4.7 ± 2.1 sec, P < 0.01). Our results suggest that administration of an appropriate dose of cannabinoid receptor agonist WIN55,212-2 has an anticonvulsant effect but does not cause respiratory depression.
  • Masako YAJIMA, Takaji YAJIMA, Tamotsu KUWATA
    2005 年 26 巻 6 号 p. 249-255
    発行日: 2005年
    公開日: 2006/01/16
    ジャーナル フリー
    Lactoferrin (LF) plays various anti-inflammatory roles in inflammation experimentally induced by lipopolysaccharides (LPS). But the effects of LF on albumin extravasation and neutrophilia have not been elucidated. We aimed to study the effects of LF on albumin extravasation, neutrophilia and/or on other symptoms in inflammation caused by LPS in rats. Human lactoferrin (hLF) was injected (10 mg/100 mL in PBS) 18 h, or 15 min prior to, or 60 min after intraperitoneal injection of LPS in 13 days old Sprague Dawley rats. Prophylactic injection of hLF significantly ameliorated albumin extravasation in ascitic fluid at 5 h and neutrophilia in the blood at 24 h after LPS injection, but the after-injection of hLF did not. Interestingly, an injection of rat anti-TNFα IgG 15 min prior to LPS injection did not ameliorate albumin extravasation. Prophylactic injection of hLF significantly ameliorated other symptoms like mortality, and the decrease of phagocytotic activity of peritoneal polymorpho-nuclear leukocytes (PMNL), but did not ameliorate the decrease of platelets in the plasma. These findings suggest that hLF may be available as a medical treatment prior to surgery for prophylaxis of side effects like albumin extravasation or neutrophilia.
  • Yuta SUZUKI, Kenichi MATSUZAKA, Ken ISHIZAKI, Masakazu TAZAKI, Toru SA ...
    2005 年 26 巻 6 号 p. 257-269
    発行日: 2005年
    公開日: 2006/01/16
    ジャーナル フリー
    The purpose of this study was to investigate the relationship between Merkel cells and nerve elements during tissue regeneration after receiving dental implants. Golden hamsters were divided into 3 groups and titanium alloy implants were fixed in their left-side maxilla through the third palatine ruga. Animals were sacrificed at 1, 2, 3, 4, 5, 6, and 7 days after the implantation and tissues were characterized at the immunohistochemical and morphological levels. CK 20 and PGP 9.5 antibodies which react with Merkel cells and nerve fibers were used. Immunohistochemically, no CK20-positive Merkel cells were seen in the peri-implant epithelium throughout the 7 days. However, starting at day 4, PGP 9.5-positive nerve fibers appeared in the connective tissue, and by day 7, nerve fibers had invaded the more superficial layer of the peri-implant epithelium compared to the mucosa removal control group. At the electron microscopic level, the intercellular spaces of the regenerating epithelium in the mucosa removal control group were small. In contrast, intercellular spaces of the peri-implant epithelium tended to be wide and regenerating nerve fibers invaded those intercellular spaces. In both the mucosa removal control group and the implantation group, the basal lamina and connective tissues regenerated completely. However, clear Merkel cells containing neurosecretory granules were not observed. Taken together, our results indicate that Merkel cells in the hamster palatine mucosa do not regenerate in the peri-implant epithelium. However, regenerative nerve fibers seem to play essential roles as part of the defense and sensory systems around the peri-implant epithelium to compensate for the weakened defense mechanism.
  • Toshihiko IWANAGA, Marie GOTO, Masahiko WATANABE
    2005 年 26 巻 6 号 p. 271-278
    発行日: 2005年
    公開日: 2006/01/16
    ジャーナル フリー
    L-Glutamate transport by intestinal epithelial cells is an initial step of the entire glutamate metabolism pathway in the gut mucosa. The present study examined the cellular distribution of glutamate transporters in the digestive tract of adult mice using immunohistochemistry and in situ hybridization technique. Expression of EAAC1 mRNA was more intense in the ileum, where the epithelium in crypts and the basal half of intestinal villi showed high levels of transcripts, suggesting an essential role of EAAC1 in differentiating or premature epithelial cells. Electron-microscopically, EAAC1 immunoreactivity was predominantly localized in the striated border of enterocytes. Immunoreactivity for GLT-1 was found in the lateral membrane of epithelial cells at the bottom of gastric glands and at the intestinal crypts, and also in the lateral membrane of secretory cells at the duodenal gland. GLAST immunoreactivity was restricted to the fundic and pyloric glands, and was especially intense in the neck portion of both glands. However, in situ hybridization analysis failed to confirm the expression of GLT-1 and GLAST at the mRNA level, possibly due to limited sensitivity. The strong and specific luminal localization of EAAC1 in the intestinal epithelium suggests that EAAC1 is a predominant transporter of glutamate, at least in the lower part of the small intestine.
  • Yoshinao HOSAKA, Hiromi UEDA, Tadatsugu YAMASAKI, Daisuke SUZUKI, Naoy ...
    2005 年 26 巻 6 号 p. 279-286
    発行日: 2005年
    公開日: 2006/01/16
    ジャーナル フリー
    The aim of this study was to investigate alterations of cultured tendon tissues to determine whether tissue culture is a useful method for biological analyses of the tendon. Tendon tissues for tissue culture were isolated from Achilles tendons of rabbits. The tendon segments were placed one segment per well and incubated in growth medium consisting of Dullbecco's modified Eagle's medium supplemented with 5% fetal bovine serum at 37°C in a humidified atmosphere with 5% CO2 for various periods. The alignment of collagen fibrils was preserved for 48 h, but tendon structure has disintegrated at 96 h. Alcian blue staining and gelatine zymography revealed that proteoglycan markedly diminished and that matrix metalloproteinase (MMPs) activity was upregulated sharply at 72 and 96 h. The ratio of collagen fibrils with large diameter had increased and the mean diameter and mass average diameter value had reached maximum at 48 h. The values then decreased and mean diameters at 72 and 96 h were significantly different from that at 48 h. At 96 h, the ratio of collagen fibrils with small diameters had increased and collagen fibrils with large diameters had disappeared. These findings indicate that structural alteration is possible to be induced by disintegration of collagen fibrils and disappearance of glycosaminoglycans from extracellular matrix (ECM), subsequent of upregulation of MMPs activity. Although the study period is limited, the tissue culture method is available for investigating cell-ECM interaction in tendons.
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