钙调蛋白在介导大鼠结肠平滑肌细胞Ca^2+内流的作用机制  被引量:2

Mechanism of Calmodulin Mediating Ca^2+ Influx in Colon Smooth Muscle Cells in Rats

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作  者:卢敏[1] 

机构地区:[1]中国医科大学附属一院外科,沈阳辽宁110001

出  处:《医学临床研究》2010年第4期577-578,581,共3页Journal of Clinical Research

摘  要:【目的】探讨钙调蛋白在介导大鼠结肠平滑肌细胞中Ca^2+内流的作用机制。【方法】将酶解分离好的大鼠结肠平滑肌细胞,随机分为以下4组:①对照组;②EGTA组;③EGTA+Veraparmil组;④EGTA+W7组。荧光探针Fura-2/AM标记细胞内游离Ca^2+,荧光分光光度计检测大鼠结肠平滑肌细胞Ca^2+浓度。【结果】在无Ca^2+缓冲液中加入EGTA(1mmol/L)使Ca^2+浓度由静息时(82.24±3.65)nmol/L升高至(267.45±4.86)nmol/L,继之,向细胞外液中引入两种浓度的Ca^2+(1.5mmol/L和3.0mmol/L),导致Ca^2+浓度进一步升高,分别为(470.23±4.21)hmol/L、(945.32±3.56)nmol/L.上述升高效应对L型电压操纵性钙通道阻滞剂维拉帕米(verapamil,5μmol/L)不敏感(P〉0.05),但钙调蛋白抑制剂W7对上述升高效应有抑制作用(P〈0.01)。【结论】钙调蛋白参与大鼠结肠平滑肌细胞的钙内流。钙调蛋白抑制剂W7能抑制由EGTA引起的细胞钙内流。这对于进一步深入研究钙通道活性改变,为预防和控制因胃肠动力紊乱所致的消化管疾病具有重要意义。[Objective]To explore the mechanism of calmodulin mediating Ca^2+ influx in colon smooth muscle cell in rats. [Methods] Colon smooth muscle cells in rats were divided into control group, EGTA group, EGTA+ verapamil group and EGTA + W7 group. Intracellular free Ca^2+ was labeled by Fura-2/AM fluorescent probe, and [Ca^2+ ] in colon smooth cells in rats was measured by fluorospectrophotometer. [Results] In the buffer of absence of Ca^2+ , EGTA(lmrnol/L) could elevate [Ca^2+] from (82.24±3.65)nmol/L to (267.45±4.86)nmol/L. And then two concentrations of Ca^2+ was added into the extracellular solution, which resulted Ca^2+ further elevated to 470.23 ±4.21)nmol/L and (945.32±3.56)nmol/L, respectively. The elevated response was blocked by W7( P 〈0.01), but was insensitive to L-type voltage calcium channel blocker veraparnil ( P 〉0. 05). [Conclusion] Calmodulin involves in Ca^2+ influx in colon smooth muscle cells. The inhibitor of calmodulin such as W7 can inhibit Ca^2+ influx caused by EGTA. It is very important to further study the change of calcium channel for preventing and control digestive disease caused by gastrointestinal mobility disorder.

关 键 词: 平滑/细胞学 结肠 钙调蛋白  大鼠 

分 类 号:R349.21[医药卫生—基础医学]

 

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