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31.
Mochida S Yoshimoto T Mimura S Inao M Matsui A Ohno A Koh H Saitoh E Nagoshi S Fujiwara K 《Biochemical and biophysical research communications》2004,317(1):114-120
Osteopontin, a crucial factor for Th1 immune response, is expressed in stellate cells and macrophages activated in injured liver. To clarify the role of osteopontin in inflammatory changes in the liver, we attempted to establish transgenic mice expressing osteopontin in hepatocytes. Mouse osteopontin cDNA, cloned from concanavalin-A-stimulated spleen cells in C57BL/6 mice, was constructed into the vector containing serum amyloid-P component promoter. This construction was microinjected into fertilized eggs of C57BL/6 mice, and 4 lines of the transgenic mice were obtained. Western blotting and immunohistochemistry revealed that osteopontin was expressed in hepatocytes, but not in non-parenchymal cells, in the transgenic mice. The mean osteopontin concentrations in the liver and plasma in the mice were 13 and 2.6 times higher than those in negative littermates. Antinuclear antibody was positive in the plasma in 50% of the transgenic mice. In the transgenic mice later than 12 weeks of age, mononuclear cell infiltration in the liver developed, and these cells were positive for CD8 and HLA-DR. Plasma ALT activity was increased with focal necrosis in hepatic lobules in the transgenic mice later than 24 weeks of age. The transgenic mice expressing osteopontin in hepatocytes may be useful as a model of autoimmune hepatitis. 相似文献
32.
Genetic polymorphims in promoter region of osteopontin gene may be a marker reflecting hepatitis activity in chronic hepatitis C patients 总被引:2,自引:0,他引:2
Mochida S Hashimoto M Matsui A Naito M Inao M Nagoshi S Nagano M Egashira T Mishiro S Fujiwara K 《Biochemical and biophysical research communications》2004,313(4):1079-1085
BACKGROUND AND AIMS: Osteopontin, an extracellular matrix protein with RGD motif, is shown to be a cytokine essential for Th1 immune response initiation. Genetic polymorphisms in the osteopontin gene (OPN) determine the magnitude of immunity against rickettsial infection in mice. Similar polymorphisms, if present also in human beings, might affect hepatitis activity in those infected with HCV. METHODS: Blood was collected from 176 patients with chronic hepatitis C. SNPs in the promoter region of OPN were analyzed in 20 patients by direct sequencing of DNA fragments amplified by PCR and in 156 patients by Invader assay. Ninety-five patients compatible to evaluation criteria were classified into three groups depending on maximal serum ALT levels during the observation periods at least for 2 years as follows; lower than 30IU/L (low-activity group), between 30 and 80IU/L with no hepatoprotective treatment (medium-activity group), and higher than 80IU/L irrespective of hepatoprotective treatment (high-activity group). RESULTS: There were 16, 19, and 60 patients in the low-, medium-, and high-activity groups, respectively. Four SNPs (nt -155, -443, -616, and -1748) were detected in the promoter region of OPN. Among them, the SNP at nt -443 (C or T) was a novel one and showed an association with hepatitis activity in our patients: T/T homozygosity was found in 2 (13%), 8 (42%), and 25 (44%), and C/T heterozygosity in 12 (75%), 8 (42%), and 23 (40%), in the low-, medium-, and high-activity groups, respectively. The other 3 SNPs already known showed linkage disequilibrium with D(') and r(2) greater than 0.937 to each other without correlation to disease activity. CONCLUSIONS. OPN promoter region SNP at nt -433 may be a useful marker reflecting hepatitis activity in chronic hepatitis C patients. 相似文献
33.
The translocation profiles of 11C-photoassimilates from eithertendrils or leaflets of the compound leaf of Pisum sativum weresimilar in shape, speed and susceptibility to blockage by chillingand heat girdling. When the feed leaf component was exposedto an anaerobic gas stream consisting of N2 gas supplementedwith 40 Pa CO2, the export of previously-fixed 11C-photoassimilatesfrom both leaflets and tendrils continued in the light, butstopped in the dark. However, in the light, translocation of11C-assimilates from the leaflet was rapidly blocked by a flowof pure N2 (i.e. anoxia). Movement of 11C-assimilates from theleaf of another C3 plant, sunflower, was similar to that fromthe pea leaflet. In contrast to both laminar leaf components,export from the tendrils was stopped under pure N2 only in thedark. Taken together the data suggest that photosynthetic O2production facilitated the movement of 11C-assimilates in theabsence of exogenous O2. The differences observed between thetendrils and the leaflets exposed to pure N2 could be attributedto the greater capacity of tendrils to produce and recycle CO2to support photosynthetic O2 production in the light. Key words: Pea, 11C-translocation, anoxia, tendril, leaflet 相似文献
34.
S Mashiba S Mochida K Ishikawa M Inao A Matsui A Ohno H Ikeda S Nagoshi M Shibuya K Fujiwara 《Biochemical and biophysical research communications》1999,258(3):674-678
Activated hepatic stellate cells produce vascular endothelial growth factor (VEGF). VEGF has been shown to act on mesenchymal cells as well. If hepatic stellate cells can express FLT tyrosine receptor family, flt-1 and KDR/flk-1, their function might be regulated by VEGF in an autocrine manner. This hypothesis was tested using hepatic stellate cells isolated from normal rats. Northern blot analysis and immunocytochemical study revealed that hepatic stellate cells cultured for 3 days on plastic dishes expressed both flt-1 and KDR/flk-1. When the culture was prolonged to 10 days, the flt-1 mRNA expression was increased, whereas both KDR/flk-1 mRNA and protein expressions diminished. DNA and collagen syntheses were minimal in the cells cultured for 3 days, but marked in those cultured for 10 days. Addition of recombinant human VEGF to the culture medium did not change both syntheses but attenuated an increase of smooth muscle alpha-actin expression in the cells during culture on plastic dishes and also contraction of collagen gels on which the cells were cultured. We conclude that VEGF may inhibit contraction of hepatic stellate cells appearing during activation by culture, probably through attenuation of smooth muscle alpha-actin expression via upregulated VEGF receptor, flt-1. 相似文献
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36.
The iron-chlorophyllin complex was found to be chemiluminescent (CL) in an acetonitrile (22%)/water mixed solvent. In the presence of hydrogen peroxide, when iron-chlorophyllin was added to the mixed solvent, a sharp CL signal immediately appeared. Also, analysis of the absorption spectra revealed decomposition of iron-chlorophyllin (based on decrease in absorbance at 396 nm), hence iron-chlorophyllin is the CL substance. Moreover, the CL intensity decreased in the presence of potassium thiocyanate (KSCN), indicating that the axial coordinative position of iron-chlorophyllin acts as a point of catalytic activation. In addition, when fluorophores were present with iron-chlorophyllin CL, their CL intensity values were similar to or greater than that of the well-known trichlorophenylperoxalate ester (TCPO) CL. Thus, during the decomposition reaction of iron-chlorophyllin, the latter transfers its energy to the coexisting fluorophores. Moreover, since the decomposed compound in this CL reaction had a fluorescence, it was found that the iron-chlorophyllin also functions as an energy donor. Therefore, the iron-chlorophyllin complex acts not only as a CL substance, but also as a catalyst and energy donor in the reaction. 相似文献
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38.
Tumor necrosis factor-alpha downregulates adrenomedullin receptors in human coronary artery smooth muscle cells 总被引:1,自引:0,他引:1
We examined the effects of tumor necrosis factor (TNF)-alpha on the expression and functionality of adrenomedullin (AM) receptors in cultured human coronary artery smooth muscle cells. Analysis of real-time quantitative polymerase chain reactions showed that these cells abundantly express two AM receptors comprised of calcitonin receptor-like receptor (CRLR) and receptor activity-modifying protein 1 (RAMP1) or RAMP2. TNF-alpha induced time- and dose-dependent decreases in the expression of CRLR and RAMP1/2 mRNAs, thereby diminishing AM-evoked cAMP production. The suppression of these three mRNAs was unaffected by inhibiting NOS, protein kinase G, protein kinase A, superoxide formation or NF-kappaB activation. 相似文献
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40.
Yusuke Kashiwagi Tomohisa Nagoshi Takuya Yoshino Toshikazu D. Tanaka Keiichi Ito Tohru Harada Hiroyuki Takahashi Masahiro Ikegami Ryuko Anzawa Michihiro Yoshimura 《PloS one》2015,10(6)