小鼠C2C12肌管中P2嘌呤受体介导的环磷酸腺苷形成的特征分析
Characterization of P2-purinoceptor mediated cyclic AMP formation in mouse C2C12 myotubes.
作者信息
Henning R H, Duin M, den Hertog A, Nelemans A
机构信息
Department of Pharmacology/Clinical Pharmacology, University of Groningen, The Netherlands.
出版信息
Br J Pharmacol. 1993 Sep;110(1):133-8. doi: 10.1111/j.1476-5381.1993.tb13782.x.
Abstract
- The formation of adenosine 3':5'-cyclic monophosphate (cyclic AMP) and inositol(1,4,5)trisphosphate (Ins(1,4,5)P3), induced by ATP and other nucleotides was investigated in mouse C2C12 myotubes. 2. ATP (100 microM) and ATP gamma S (100 microM) caused a sustained increase in cyclic AMP content of the cells, reaching a maximum after 10 min. The cyclic AMP content reached a maximum in the presence of 100 microM ATP, followed by a decline at higher ATP concentrations. ATP-induced cyclic AMP formation was inhibited by the P2-purinoceptor antagonist, suramin. 3. Myotubes hydrolysed ATP to ADP at a rate of 9.7 +/- 1.0 nmol mg-1 protein min-1. However, further hydrolysis of ADP to AMP and adenosine was negligible. 4. The cyclic AMP formation induced by ADP (10 microM-1 mM) showed similar characteristics to that induced by ATP, but a less pronounced decline was observed than with ATP. ADP-induced cyclic AMP formation was blocked by suramin, while cyclic AMP formation elicited by adenosine (10 microM-1 mM) was insensitive to suramin. 5. The ATP analogue, alpha,beta-methylene-ATP also induced a suramin-sensitive cyclic AMP formation, while 2-methylthio-ATP and the pyrimidine, UTP, did not affect cyclic AMP levels. 6. Stimulation of the myotubes with ATP or UTP (10 microM-1 mM) caused a concentration-dependent increase in the Ins(1,4,5)P3 content of the cells. ADP (100 microM-1 mM) was less effective. Adenosine did not affect Ins(1,4,5)P3 levels. 7. Incubation of the cells with UTP (30 microM- 1 mM) inhibited the ATP- and ADP-induced cyclic AMP formation, suggesting that stimulation of the 'nucleotide' type P2-receptor inhibits P2-purinoceptor mediated cyclic AMP formation in C2C12 myotubes. In contrast, UTP (30 microM-I mM) enhanced adenosine-induced cyclic AMP formation.8. Adenosine-sensitive P1-purinoceptors activating cyclic AMP formation were found in C2C12 myotubes.Further, a novel P2-purinoceptor is postulated, sensitive to ATP, ADP and ATPgammaS, which also activates the formation of cyclic AMP in C2C12 myotubes.
摘要
- 研究了ATP和其他核苷酸在小鼠C2C12肌管中诱导生成3':5'-环磷酸腺苷（环磷酸腺苷）和肌醇(1,4,5)三磷酸（Ins(1,4,5)P3）的情况。2. ATP（100微摩尔）和ATPγS（100微摩尔）使细胞内环磷酸腺苷含量持续增加，10分钟后达到最大值。在100微摩尔ATP存在的情况下，环磷酸腺苷含量达到最大值，在更高的ATP浓度下随后下降。ATP诱导的环磷酸腺苷生成受到P2嘌呤受体拮抗剂苏拉明的抑制。3. 肌管将ATP水解为ADP的速率为9.7±1.0纳摩尔/毫克蛋白质·分钟。然而，ADP进一步水解为AMP和腺苷的量可忽略不计。4. ADP（10微摩尔至1毫摩尔）诱导的环磷酸腺苷生成与ATP诱导的具有相似特征，但与ATP相比，下降不太明显。ADP诱导的环磷酸腺苷生成被苏拉明阻断，而腺苷（10微摩尔至1毫摩尔）诱导的环磷酸腺苷生成对苏拉明不敏感。5. ATP类似物α,β-亚甲基-ATP也诱导了对苏拉明敏感的环磷酸腺苷生成，而2-甲硫基-ATP和嘧啶UTP不影响环磷酸腺苷水平。6. 用ATP或UTP（10微摩尔至1毫摩尔）刺激肌管导致细胞内Ins(1,4,5)P3含量呈浓度依赖性增加。ADP（100微摩尔至1毫摩尔）的效果较差。腺苷不影响Ins(1,4,5)P3水平。7. 用UTP（30微摩尔至1毫摩尔）孵育细胞抑制了ATP和ADP诱导的环磷酸腺苷生成，表明刺激‘核苷酸’型P2受体抑制了C2C12肌管中P2嘌呤受体介导的环磷酸腺苷生成。相反，UTP（30微摩尔至1毫摩尔）增强了腺苷诱导的环磷酸腺苷生成。8. 在C2C12肌管中发现了激活环磷酸腺苷生成的腺苷敏感型P1嘌呤受体。此外，推测存在一种新型P2嘌呤受体，对ATP、ADP和ATPγS敏感，它也能激活C2C12肌管中环磷酸腺苷的生成。
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