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标题：Involvement of M3 Muscarinic Receptors in ACh-Induced Increase in Membrane-Associated RhoA of Rat Bronchial Smooth Muscle
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作者：Hiroyasu Sakai ; Tsuyoshi Shirai ; Maki Yamamoto 等
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：2005
卷号：28
期号：4
页码：625-628
DOI：10.1248/bpb.28.625
出版社：The Pharmaceutical Society of Japan
摘要：It is known that RhoA is translocated from cytoplasm to cell membrane in bronchial smooth muscle when activated by acetylcholine (ACh) stimulation. In the present study, the effects of selective muscarinic receptor antagonist methoctramine, AF-DX116 (for M₂) and 4-diphenylacetoxy N -methylpiperidine (4-DAMP; for M₃) on the ACh-induced rat bronchial smooth muscle contraction and increase in membrane-associated RhoA were investigated to elucidate the muscarinic receptor subtype participating in these responses. To evaluate ACh-induced contraction of bronchial smooth muscle, bronchial ring of rat was prepared, suspended in an organ bath and the tension was measured isometrically. To quantify the ACh-induced increase in membrane-associated RhoA protein, western blot analysis was performed by using homogenates of membrane and cytosolic fractions of the rat bronchi. The muscarinic M₂ and M₃ receptors were detected by using RT-PCR in rat bronchial smooth muscle. Both the ACh-induced smooth muscle contraction and increase in membrane-associated RhoA were markedly inhibited by 4-DAMP, but not by methoctramine or AF-DX116. In conclusion, these results indicated contraction for the first time that the activation of RhoA occurs via M₃ receptor in rat bronchial smooth muscle.
关键词：bronchial smooth muscle;acetylcholine;muscarinic receptor;RhoA