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Intracellular signalling pathway involved in ATP release from isolated smooth muscle cells.

细胞内信号通路参与分离的平滑肌细胞释放 ATP。

基本信息

批准号：
07670127
负责人：
KATSURAGI Takeshi
金额：
$ 1.22万
依托单位：
Fukuoka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670127/
关键词：
ATP release Cultured ileal smooth muscle cells Intracellular signalling Inositol 1,4,5-trisphosphate alpha, beta-Methylene ATP LiCl Neomycin Pertussis toxin P_2-受容体アゴニスト M-受容体アゴニスト ATP放出イノシトール1,4,5-三リン酸生成フォスフォリパーゼC LiCI モルモット

项目摘要

In recent investigations, ATP is known to exhibit a wide-variety of cell responses mediated by activation of P2X and P2Y-purinoceptors and, thus, seems to play roles of mediators to transfer signals to "cell to cell".In a series of our studies, it has been persented findings that stimulation of transmitters' receptors with noradrenaline and ACh elicits ATP release from smooth muscle segments.During the first year (1995) of the project, ATP released to superfusate from cultured smooth muscle cells from ileum and taenia coli of guinea-pig was measured by the luciferase method. The release of ATP was increased in the presence of alpha, beta-methylene ATP,bethanechol and peptides such as angiotensin II.The increased release was attenuated by respective antagonists. Accordingly, the findings suggest that ATP release comes from the smooth muscle cells.In 1996, the research project was focused on determination of inositol 1,4,5-trisphosphate (IP3) accumulation from the homogenate of ileal longitudinal muscles in the persence and absence of alpha, beta-methylene ATP.IP3 was measured by a radio-receptor assay using a IP3 assay kit (Amersham, TRK 1000). The P2-agonist enhanced the accumulation of IP3 in the homogenate. The enhanced accumulation was inhibited by blockade of phospholipase C and phosphoinositide turnover with neomycin and Li^+, respectively. The release of ATP evoked by alpha, beta-methylene ATP was insensitive to pertussis toxin and was counteracted by neomycin and spermine and by Li^+.Taken together, it is concluded that the enhancement of IP3 accumulation may be involved in the evoked release of ATP.

在最近的研究中，已知ATP表现出由P2 X和P2 Y-嘌呤受体的激活介导的多种细胞应答，因此，ATP似乎在“细胞到细胞”的信号传递中起介质的作用。在第一年（1995年），用去甲肾上腺素（NE）和乙酰胆碱（ACh）刺激递质受体可促进平滑肌ATP的释放。本课题采用荧光素酶法测定豚鼠回肠和结肠带平滑肌细胞灌流液中ATP的释放量。在α，β-亚甲基ATP，氨甲酰甲胆碱和肽如血管紧张素II的存在下，ATP的释放增加。在1996年，研究项目集中于测定在存在和不存在α，β-亚甲基ATP的情况下回肠纵肌匀浆中1，4，5-三磷酸肌醇（IP 3）的积累。使用IP 3测定试剂盒（阿默舍姆，TRK 1000）通过放射受体测定来测量IP 3。P2-激动剂增强了匀浆中IP 3的积累。分别用新霉素和Li^+阻断磷脂酶C和磷脂酰肌醇的周转，可抑制这种增加的积累。α，β-亚甲基ATP诱发的ATP释放对百日咳毒素不敏感，但能被新霉素、精胺和Li^+所抵消，提示ATP的诱发释放可能与IP 3积累增加有关。