Mechamism on amylase secretion

淀粉酶分泌机制

• 批准号: 04671139
• 负责人: SHIMOMURA Hiromi
• 金额: $ 1.34万
• 依托单位: The Nippon Dental University, Niigata
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04671139/
• 关键词: ANP; ANP receptor; amlase; cAMP; cGMP; adenylate cyclase; forskolin; Gi protein; Rat salivary gland; cyclic AMP; cyclic GMP; Forskolin

Roles of cGMP on salivary secretion were investigated using atrial natriuretic peptide (ANP) which is known as activator of quanylate cyclase.1) The amount of ^<125>I-ANP specific binding to plasma membrane of rat parotid and submandibular gland increased in proportion to the amount of protein used and reached to plateau. ANP competed binding of ^<125>I-ANP with the membrane protein, and IC_<50> values for parotid and submandibular gland membrane were 5 X 10^<19>M and 10^<19>M, respectively. The results indicated the presence of ANP receptor protein in the salivary glands.2) Rat parotid acinar cells were incubated with ANP or carbachol.Caubachol (10^<-5>M) had no detectable effect on cGMP accumulation (only 5%). However, ANP(10^<-7>M) enhanced cGMP levels with an approximately 2-fold increase. ANP alone didn't cahange cAMP accumulation, but inhibited cAMP accumulation stimulated by forskolin.ANP, nitroprusside (NP) and hydroxylamine (HA) enhanced cGMP in the presence or absence of forskolin. ANP inhibited cAMP accumulation stimulated by forskolin, but NP and HA did not affect. Further more, ANP inhibited amylase resease from rat parotid acinar cells stimulated by forskolin, di-Bu-cAMP and isoproterenol, but NP and HA didn't inhibite. The results indicated cGMP does not mediate cAMP accumulation and amylase secretion directly.4) The inhibition of amylase release and cAMP accumulation by ANP was inhibited by treatment of parotid gland or plasma membrane of parotid gland with pertussis toxin (PT). Treatment of membrane with PT afforded ribosylated protein. The results suggested that the inhibition of adenylate cyclase by ANP may be mediated by Gi protein through ANP clearance receptor.5) To investigate the effect of cGMP on cGMP-stimulated phosphodiesterase (PED), identifications of PDEs were performed using specific PDE-inhibitors and estimation of molecular weight of PDEs in rat parotd gland. Type I, II, III and IV cAMP-specific-PDEs were identified.

本实验用心房钠尿肽（ANP）研究了cGMP对唾液分泌的影响。1）<125>大鼠腮腺和下颌下腺细胞膜上的~ 1H-ANP特异性结合量随蛋白质用量的增加而增加，并达到平台期。ANP<125>与膜蛋白竞争结合，<50>对腮腺和下颌下腺膜的IC_（1/2）值分别为5 × 10 ~（-1）<19>M和10 ~<19>（-1）M。结果表明，大鼠腮腺腺泡细胞中存在ANP受体蛋白。2）将大鼠腮腺腺泡细胞与ANP或卡巴胆碱共同孵育，10 μ <-5>M的卡巴胆碱对cGMP的积累无明显影响（仅5%）。然而，ANP（10 μ <-7>M）增强cGMP水平，增加约2倍。单独使用ANP不影响cAMP的积累，但可抑制forskolin刺激的cAMP的积累，在forskolin存在或不存在的情况下，ANP、硝普钠（NP）和羟胺（HA）均促进cGMP的积累。ANP抑制forskolin刺激的cAMP积累，而NP和HA则无此作用。此外，ANP还能抑制forskolin、di-Bu-cAMP和异丙肾上腺素刺激的大鼠腮腺腺泡细胞淀粉酶的释放，而NP和HA则无此作用。结果表明cGMP不直接介导cAMP的积累和淀粉酶的分泌。4）用百日咳毒素（PT）处理腮腺或腮腺质膜，可抑制ANP抑制淀粉酶释放和cAMP积累的作用。用PT处理膜得到核糖基化蛋白。结果提示，ANP对腺苷酸环化酶的抑制作用可能是由Gi蛋白通过ANP清除受体介导的。5）为了研究cGMP对cGMP刺激的磷酸二酯酶（PED）的影响，采用特异性PDE抑制剂对大鼠腮腺PDE进行了鉴定，并估算了PDE的分子量。鉴定了I、II、III和IV型cAMP特异性PDE。

项目成果

梨田智子: "唾液腺Atrial Natriuretic peptide(ANP)受容体とサイクリックヌクレオチドの濃度変化" 歯科基礎医学会雑誌(補冊). 34. 123- (1992)

Tomoko Nashida：“唾液腺心房钠尿肽（ANP）受体和环核苷酸浓度的变化”日本基础牙科医学会杂志（增刊）34. 123-（1992）。

屋部朋子: "ラット唾液腺Atrial Natriuretic peptide(ANP)受容体に関する研究" 歯学. 80. 1441-1449 (1993)

Tomoko Yabe：“大鼠唾液腺心房钠尿肽（ANP）受体的研究”牙科。 80. 1441-1449（1993）

梨田 智子、 今井 あかね 下村 浩巳: "唾液線 Atrial Natriuretic Peptide(ANP) 受容体とサイクリックヌクレオチドの濃度変化" 歯科基礎医学会雑誌(Supplement). 34. 123 (1992)

Tomoko Nashida、Akane Imai、Hiromi Shimomura：“唾液腺心房钠尿肽（ANP）受体和环核苷酸浓度的变化”日本基础牙科医学会杂志（增刊）34. 123（1992）。

今井あかね、梨田智子、下村浩巳: "ラット唾液腺プロテインキナーゼに対するサイクリックヌクレオチドの働き" 歯学. 81. 397 404 (1993)

Akane Imai、Tomoko Nashida、Hiromi Shimomura：“环核苷酸对大鼠唾液腺蛋白激酶的功能”牙科。 81. 397 404 (1993)

今井あかね、梨田智子、下村浩巳: "心房性ナトリウム利尿ペプチド(ANP)による唾液腺アデニレートシクラーゼ活性の抑制とcGMPの働き" 歯科基礎医学会誌(補冊). 35. 82 (1993)

Akane Imai、Tomoko Nashida、Hiromi Shimomura：“心房钠尿肽（ANP）对唾液腺腺苷酸环化酶活性的抑制和 cGMP 的功能”日本基础牙科医学会杂志（增刊）35. 82（1993）。