Identification of ghrelin in human saliva:: Production by the salivary glands and potential role in proliferation of oral keratinocytes
Identification of ghrelin in human saliva:: Production by the salivary glands and potential role in proliferation of oral keratinocytes
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DOI：
10.1373/clinchem.2004.040667
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发表时间：
2005-06-01
影响因子：
9.3
通讯作者：
Rauh, M
中科院分区：
文献类型：
--
作者：
Gröschl, M;Topf, HG;Rauh, M
背景资料：我们调查是否ghrelin是存在于人类唾液中，是由唾液腺，和生理后果，这些findings.Methods：ghrelin和特异性受体mRNA的表达通过PCR测定。通过免疫印迹和尺寸排阻快速蛋白液相色谱法（FPLC）与连续RIA鉴定蛋白质。特异性RIA用于定量唾液总生长素和生物活性生长素释放肽。通过免疫组织化学方法研究了唾液腺冷冻切片中ghrelin的分布。Ghrelin对5-溴-2 '-脱氧尿苷掺入作为细胞增殖的量度的影响在原代口腔角质形成细胞中进行了研究。通过免疫印迹和唾液的FPLC分级分离鉴定唾液和腺体中的激素。免疫组化显示ghrelin分布在唾液腺。该受体也由腺体和口腔角质形成细胞产生，并显示出功能性。比较健康个体（体重指数18-27 kg/m2）的总ghrelin值，发现唾液中ghrelin浓度显著低于血清（P < 0.01）。两种基质之间的相关性为r（2）= 0.56（P < 0.001），与体重指数呈负相关（r（2）= 0.314; P < 0.01）。生物活性酰化生长素释放肽也存在于唾液中。与生长素释放肽孵育的角质形成细胞的细胞增殖显着增加（P < 0.001）。结论：唾液中Ghrelin由唾液腺产生并释放，其抑制剂为NOX-B11（50 nmol/L）。生长素释放肽对口腔细胞增殖的影响增加了其他唾液生长因子的促增殖作用。(c)2005年美国临床化学协会
Background: We investigated whether ghrelin is present in human saliva, is produced by salivary glands, and physiologic consequences of these findings.Methods: Expression of ghrelin and specific receptor mRNA was determined by PCR. Proteins were identified by immunoblotting and size-exclusion fast protein liquid chromatography (FPLC) with consecutive RIA. Specific RIAs were used for quantification of salivary total and bioactive ghrelin. Distribution of ghrelin was investigated by immunohistochemistry in cryosections of the salivary glands. The effect of ghrelin on incorporation of 5-bromo-2'-deoxyuridine as a measure of cell proliferation was investigated in primary oral keratinocytes.Results: Ghrelin is produced by the salivary glands. The hormone was identified in saliva and glands by immunoblotting and by FPLC fractionation of saliva. Immunohistochemistry demonstrated ghrelin distribution in the salivary glands. The receptor was also produced by the glands and by oral keratinocytes and was shown to be functional. Comparison of total ghrelin values for healthy individuals (body mass index, 18-27 kg/m(2)) showed significantly lower concentrations in saliva than in serum (P < 0.01). The correlation between both matrices was r(2) = 0.56 (P < 0.001) with a negative correlation to body mass index (r(2) = 0.314; P < 0.01). Bioactive acylated ghrelin was also present in saliva. Incubation of keratinocytes with ghrelin led to significantly increased cell proliferation (P < 0.001). This effect could be completely suppressed by co-incubation with NOX-B11 (50 nmol/L), a novel specific inhibitor of acylated ghrelin.Conclusions: Ghrelin in saliva is produced and released by salivary glands. The effect of ghrelin on oral cell proliferation adds to the pro-proliferative action of other salivary growth factors. (c) 2005 American Association for Clinical Chemistry