摘要
目的:通过检测大鼠结肠cajal细胞中库容性Ca2+内流(capacitative Ca2+entry,CCE),探讨导致大鼠结肠cajal细胞内Ca2+稳态失衡的信号途径,为治疗相关疾病提供新的思路。方法:荧光探针Fu-ra-2/AM标记细胞内游离Ca2+后,用荧光分光光度计检测乙二醇-双(2-氨基乙基醚)四乙酸(EGTA)耗竭胞内钙库后对分离的大鼠结肠cajal细胞Ca2+浓度([Ca2+])的影响。结果:在无Ca2+缓冲液中加入EGTA(10mmol/L),细胞内[Ca2+]由静息时的(69.37±3.02)nmol/L升高至(272.52±5.21)nmol/L;继之,向细胞外液中引入1.5mmol/L和3.0mmol/L的氯化钙溶液,导致细胞内Ca2+进一步升高为(466.43±4.63)nmol/L和(977.43±3.27)nmol/L;且此升高效应对维拉帕米(5μmol/L)不敏感,但可被2APB(20～100μmol/L)抑制。结论:酶解法并结合密度离心法分离的大鼠结肠cajal细胞上存在胞内钙库耗竭激活的Ca2+内流。这对于研究钙通道活性改变、预防和控制因胃肠动力紊乱所致的消化系统疾病具有重要意义。
Objective： To investigate the changes of capacitative Ca^2＋ entry（CCE） in colon cajal cells and its mechanism, Methods.. Intracellular Ca^2＋ changes induced by EGTA -activated Ca^2＋ influx were measured in colon cajal cells which was freshly isolated from colon of Wistar rats with acute enzymatic dissociation and ficolt density centrifugation method.with the fluorescent indicator Fura-2/AM. Results： In the absence of external Ca^2＋, the concentration of calcium-chelating agent EGTA elevated Ca^2＋ from （69.37 ±3.02）nmot/L to （272.52 ± 5.21 ）nmol/L,a subsequent reintroduction of Ca^2＋ into the extracellular solution resulted in further Ca^2＋ elevating to 466.43 ± 4.63）nmol/L and （977.43 ±3.27）nmol/L respectively, and the response was blocked by 2APB, but it was insensitive to Calcium channels blocker veraparnil .Conclusion： Ca^2＋ influx activated by store depletion are present in colon cajai Cells; it has the value for further study on gastric motility disorder diseases.
出处
《中国现代普通外科进展》
CAS
2009年第6期463-465,共3页
Chinese Journal of Current Advances in General Surgery
