{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,27]],"date-time":"2026-04-27T14:09:58Z","timestamp":1777298998237,"version":"3.51.4"},"reference-count":0,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2004,5,4]],"date-time":"2004-05-04T00:00:00Z","timestamp":1083628800000},"content-version":"vor","delay-in-days":246,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Experimental Physiology"],"published-print":{"date-parts":[[2003,9]]},"abstract":"<jats:p>The Na<jats:sup>+<\/jats:sup>\u2010K<jats:sup>+<\/jats:sup>\u20102Cl<jats:sup>\u2212<\/jats:sup> cotransporter (NKCC1) is responsible for ion transport across the secretory and absorptive epithelia, the regulation of cell volume, and possibly the modulation of cell growth and development. It has been reported that a variety of cells, including osteoblasts, contain this cotransporter. In this study, the physiological role of NKCC1 in osteoclastogenesis was exploited in a co\u2010culture system. Bumetanide, a specific inhibitor of NKCC1, reduced the number of tartrate\u2010resistant acid phosphatase (TRAP)\u2010positive multinucleated cells. In order to investigate the mechanism by which bumetanide inhibits osteoclastogenesis, the mRNA expressions of the receptor activator of nuclear factor (NF)\u2010\u03baB ligand (RANKL) and osteoprotegerin (OPG) were analysed by RT\u2010PCR. Exposure of osteoblastic cells to a medium containing 1 \u00b5M bumetanide reduced RANKL mRNA expression induced by 10 nM 1\u03b1,25\u2010dihydroxyvitamin D<jats:sub>3<\/jats:sub> (1\u03b1,25(OH)<jats:sub>2<\/jats:sub>D<jats:sub>3<\/jats:sub>, in a dose\u2010dependent manner. In addition, RANKL expression was also analysed with enzyme\u2010linked immunosorbant assay (ELISA) using anti\u2010RANKL antibody. The expression of RANKL was decreased with the increase of bumetanide concentration. In contrast, the expression of OPG mRNA, a novel tumour necrosis factor (TNF) receptor family member was increased in the presence of bumetanide. These results imply that bumetanide inhibits osteoclast differentiation by reducing the RANKL\/OPG ratio in osteoblastic cells. However, no significant difference in M\u2010CSF mRNA expression was observed when bumetanide was added. Also, we found that the phosphorylation of c\u2010Jun NH<jats:sub>2<\/jats:sub>\u2010terminal kinase (JNK), which regulates the activity of various transcriptional factors, was reduced by bumetanide treatment. Conclusively, these findings suggest that NKCC1 in osteoblasts has a pivotal role in 1\u03b1,25(OH)<jats:sub>2<\/jats:sub>D<jats:sub>3<\/jats:sub>\u2010induced osteoclastogenesis partly via the phosphorylation of JNK.<\/jats:p>","DOI":"10.1113\/eph8802558","type":"journal-article","created":{"date-parts":[[2003,9,5]],"date-time":"2003-09-05T09:25:20Z","timestamp":1062753920000},"page":"569-574","source":"Crossref","is-referenced-by-count":10,"title":["Bumetanide, the Specific Inhibitor of Na<sup>+<\/sup>\u2010K<sup>+<\/sup>\u20102Cl<sup>\u2212<\/sup> Cotransport, Inhibits 1\u03b1,25\u2010Dihydroxyvitamin D<sub>3<\/sub>\u2010Induced Osteoclastogenesis in a Mouse co\u2010culture System"],"prefix":"10.1113","volume":"88","author":[{"given":"Hyun\u2010A","family":"Lee","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hyunjoo","family":"Jeong","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eun\u2010Young","family":"Kim","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mi Young","family":"Nam","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yun\u2010Jung","family":"Yoo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeong\u2010Taeg","family":"Seo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dong Min","family":"Shin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seung\u2010Ho","family":"Ohk","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Syng\u2010Ill","family":"Lee","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2004,5,4]]},"container-title":["Experimental Physiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1113%2Feph8802558","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1113%2Feph8802558","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/physoc.onlinelibrary.wiley.com\/doi\/pdf\/10.1113\/eph8802558","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,11,17]],"date-time":"2023-11-17T19:21:33Z","timestamp":1700248893000},"score":1,"resource":{"primary":{"URL":"https:\/\/physoc.onlinelibrary.wiley.com\/doi\/10.1113\/eph8802558"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2003,9]]},"references-count":0,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2003,9]]}},"alternative-id":["10.1113\/eph8802558"],"URL":"https:\/\/doi.org\/10.1113\/eph8802558","archive":["Portico"],"relation":{},"ISSN":["0958-0670","1469-445X"],"issn-type":[{"value":"0958-0670","type":"print"},{"value":"1469-445X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2003,9]]}}}