首页> 外文期刊>Biochimica et Biophysica Acta. General Subjects >Effect of bitter compounds on amylase secretion in murine submandibular glands: Signaling pathway mechanisms.
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Effect of bitter compounds on amylase secretion in murine submandibular glands: Signaling pathway mechanisms.

机译:苦味化合物对鼠下颌腺淀粉酶分泌的影响:信号通路机制。

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BACKGROUND: Amylase is synthesized in submandibular glands (SMG) and released into the oral cavity to degrade carbohydrates in the mouth. Bitter taste receptors (T2R) belong to the G-protein coupled receptor (GPCR) family and are expressed in the taste cells and also in the digestive tract. METHODS: The activity of amylase secreted by murine SMG was measured, detecting maltose by Bernfeld's method. Amylase and T2R6 were detected by imunohistochemistry and Western blot. The expression of Ggustducin, Gi, and phospholipase Cbeta2 was also studied by Western blot. cAMP levels were measured by radioimmunoassay and inositol monophosphate production was quantified by ELISA. RESULTS: Theophylline, denatonium and cycloheximide exerted a dose-dependent inhibition on amylase secretion. This effect was reverted by preincubating SMG with an anti-Galphai antibody. cAMP production was increased by the same compounds, an effect that was also abrogated by an anti-Galphai antibody. Bitter compounds reduced inositol monophosphate formation in SMG and H-89, a protein kinase A inhibitor, reverted this action, revealing that this protein kinase down regulates phospholipase C activity. GENERAL SIGNIFICANCE: We demonstrated that theophylline, denatonium and cycloheximide inhibit salivary amylase secretion, activating an intracellular signaling pathway that involves cAMP and phospholipase C, that cross talks via protein kinase A.
机译:背景:淀粉酶在下颌下腺(SMG)中合成,并释放到口腔中以降解口腔中的碳水化合物。苦味觉受体(T2R)属于G蛋白偶联受体(GPCR)家族,在味觉细胞和消化道中表达。方法:测定小鼠SMG分泌的淀粉酶的活性,用Bernfeld法检测麦芽糖。通过免疫组织化学和蛋白质印迹检测淀粉酶和T2R6。还通过Western印迹研究了古gust丁素,Gi和磷脂酶Cbeta2的表达。通过放射免疫测定法测量cAMP水平,并通过ELISA量化肌醇单磷酸的产生。结果:茶碱,地那铵和环己酰亚胺对淀粉酶的分泌具有剂量依赖性。通过将SMG与抗Galphai抗体进行预孵育可以恢复这种效果。相同的化合物可增加cAMP的产生,抗Galphai抗体也可消除这种作用。苦味化合物减少了SMG和蛋白激酶A抑制剂H-89中肌醇单磷酸酯的形成,从而恢复了该作用,表明该蛋白激酶下调了磷脂酶C的活性。一般意义:我们证明了茶碱,地那铵和环己酰亚胺可抑制唾液淀粉酶的分泌,从而激活涉及cAMP和磷脂酶C的细胞内信号传导途径,并通过蛋白激酶A相互干扰。

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