著者

Satoko Iwasa Hirotaka Ota Kensuke Nishio Mariko Ohtsu Masafumi Kusunoki Takahiro Gojoubori Tetsuo Shirakawa Masatake Asano

出版者

日本大学歯学部

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

vol.58, no.3, pp.317-323, 2016 (Released:2016-09-25)

参考文献数

25

被引用文献数

4

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Toll-like receptors (TLR) recognize microbe-associated molecular patterns and induce the innate immune response. Among them, TLR5 recognizes the Gram-negative bacterial component flagellin. The aim of this study was to examine the expression of TLR5 in mouse salivary gland (SG). The SG was excised from 8- to 10-week-old female C57BL/6 mice. Salivary gland epithelial cells (SGECs) were purified and subjected to reverse transcription polymerase chain reaction (RT-PCR). Western blotting was performed to detect TLR5 expression at the protein level in several organs. The localization of TLR5 in SG was examined using immunohistochemical staining. The responsiveness of SGECs to flagellin was further examined by evaluating the induction of CXCL1 by real-time PCR and immunoprecipitation followed by Western blotting. TLR5 expression in SG was confirmed at the gene and protein levels. Immunohistochemical staining detected TLR5 in both acinic and ductal cells of the sublingual gland, but not in serous acinic cells of the submandibular gland. Although TLR5 was detected throughout the cytoplasm in ductal cells, positive staining was observed on the basal side of the mucous acinic cells. The purified SGECs responded to flagellin and induced the production of CXCL1. These findings suggest that TLR5 is functionally expressed in the SG and responds to its cognate ligand flagellin. (J Oral Sci 58, 317-323, 2016)

著者

Eri Sata Leo Takada Ryo Kaetsu Mai Fukasawa Mariko Ohtsu Mitsuru Motoyoshi Masatake Asano

出版者

Nihon University School of Dentistry

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

pp.19-0477, (Released:2020-06-04)

参考文献数

14

被引用文献数

3

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Interleukin-1α (IL-1α) is produced inside cells in its precursor form (pIL-1α). Enzymatic cleavage yields mature (mIL-1α) and the propiece of IL-1α (ppIL-1α), which are thought to be localized in the nucleus, because of the presence of nuclear localizing signals. Studies of ppIL-1α function have been hampered by the lack of a ppIL-1α-specific antibody (Ab). In the present study, the authors generated anti-ppIL-1α Ab by using recombinant histidine-tagged ppIL-1α (His-ppIL-1α) as an immunogen. Rabbits were immunized with His-ppIL-1α, and affinity-purified Ab was obtained. Ab reactivity and specificity were examined by Western blotting. The antibody successfully recognized transfectant-derived green fluorescence protein (GFP)-tagged ppIL-1α but not GFP. A sandwich enzyme-linked immunosorbent assay (ELISA) system established by biotinylating the anti-ppIL-1α Ab successfully detected GFP-ppIL-1α. The Ab and ELISA system allows functional analysis of ppIL-1α and improves understanding of ppIL-1α.

著者

Satsuki Saito Mariko Ohtsu Masatake Asano Tomohiko Ishigami

出版者

Nihon University School of Dentistry

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

pp.18-0411, (Released:2019-09-20)

参考文献数

24

被引用文献数

4

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Accurate evaluation of the anti-cancer effects of ouabain, a cardiac glycoside, requires an understanding of its signaling pathway. This study examined the effects of ouabain stimulation on spontaneous interleukin (IL)-8 and IL-1α secretion in the HSC3 oral squamous cell carcinoma cell line. IL-8 secretion was reduced and IL-1α secretion was increased in the cells. Real-time polymerase chain reaction confirmed that these changes were regulated at the transcriptional level. Further analysis revealed that ouabain stimulation induced phosphorylation of activator protein (AP)-1 components (c-Jun and c-Fos) but not nuclear factor kappa B (NF-κB) components (p65 and p50). A luciferase assay demonstrated that the NF-κB-binding site located at 1 kb upstream of the TATA box in the IL-8 gene contributed to the reduction in IL-8 secretion. Pre-incubation of the cells with BAPTA-AM and L-glutathione increased IL-8 secretion, which indicates that Ca2+ ions and reactive oxygen species are associated with the ouabain-mediated reduction in IL-8 levels. The inhibitory effect of ouabain was attributed to reduced nuclear translocation of the NF-κB p65 subunit. Taken together, these findings indicate that ouabain exerts opposing effects on transcription factors NF-κB and AP-1.

著者

Mariko Ohtsu Muneaki Tamura Hideto Sasaki Shuichi Sato Masatake Asano

出版者

日本大学歯学部

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

vol.59, no.3, pp.453-456, 2017 (Released:2017-09-14)

参考文献数

16

被引用文献数

1

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The effects of bittern water (BW), obtained from the ocean floor, on cariogenic bacteria and saliva secretion were examined. Streptococcus mutans was mixed with BW for 1, 3, 5, 10, and 20 min to explore the bactericidal effects of BW against cariogenic bacteria. Bacterial viability was calculated by counting the number of colony-forming units on Brain Heart Infusion agar plates. The results indicated a bacterial viability of more than 35% even after 20 min of incubation. Subsequently, the effects of BW on saliva secretion and the salivary concentration of secretory IgA (sIgA) were examined. Gargling with BW significantly augmented saliva secretion. Although the sIgA concentration was reduced, the total sIgA secreted into saliva was increased significantly. Our findings indicate that the use of BW may be a new strategy for the treatment of various oral diseases, including dry mouth.

著者

Saeka Mochizuki Mayuko Takano Naoyuki Sugano Mariko Ohtsu Kou Tsunoda Ryosuke Koshi Naoto Yoshinuma

出版者

日本酸化ストレス学会

雑誌

Journal of Clinical Biochemistry and Nutrition (ISSN:09120009)

巻号頁・発行日

vol.58, no.1, pp.64-68, 2016 (Released:2016-01-01)

参考文献数

33

被引用文献数

10

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The aim of this study was to test the effects of B-group vitamin supplements on wound healing in diabetic mice. The mice in the experimental group were treated daily with 1 g/L B6, 1.25 mg/L B12, and 62.5 mg/L folic acid in their drinking water. Full-thickness excision wounds were created with 6-mm skin biopsy punches. Each wound closure was digitally photographed. Beginning on day 3 after wounding, the wound area in the diabetic mice was statistically larger than that of normal mice (p<0.05 vs diabetic mice). The diabetic mice treated with B vitamins displayed accelerated wound closure on day 3 (wound area 42.8 ± 11.3%, p<0.05). On day 9 after wounding, the wound area in the diabetic mice was also statistically larger than that of normal mice (p<0.05 vs diabetic mice). The diabetic mice treated with B vitamins displayed accelerated wound closure on day 3 (wound area 13.2 ± 16.8%, p<0.05). In addition, the high glucose level in the diabetic animals decreased significantly in response to B vitamin treatment. In conclusion, the results of this study indicate that B vitamin supplementation may improve wound healing in diabetic mice.