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Cajal细胞调节豚鼠胆囊动力的实验研究
徐龙1,余保平2*,陈灵丹2,吕农华1,朱萱1,陈幼祥1,余保平
0
(1.南昌大学第一附属医院消化内科,南昌 330006; 2.武汉大学人民医院消化内科,武汉 430060)
摘要:
目的证实豚鼠胆囊Cajal间质细胞(ICC)是胆囊平滑肌动力的起搏细胞。方法采用在体记录和组织钳细胞内记录的方式,研究胆囊自发电活动。亚甲蓝+光照的方法破坏胆囊ICC。胶原酶分离方法分离胆囊平滑肌细胞,并对分离的单个细胞进行形态学分析,联合激光共聚焦显微镜检测c-kit蛋白表达的方法鉴定ICC。膜片钳用于证明胆囊ICC能产生具有起搏活性的自发去极化电流。结果豚鼠胆囊在体记录的慢波每分钟(58±3.2)次,细胞内记录时可记录到慢波和动作电位。采用亚甲蓝+光照的方法后,慢波和动作电位明显减弱或消失。豚鼠胆囊新鲜分离的细胞经过贴壁之后,可见少量具有典型ICC光镜特征的细胞,占细胞总数的(5±1.2)%(n=12),(21±4)%细胞的c-kit蛋白表达阳性。全细胞膜片钳记录胆囊ICC的自发去极化电流,发现豚鼠胆囊ICC自发去极化电流频率每分钟(58.4±3.5)次 (n=21),幅度约-(72±3.5) pA。结论豚鼠胆囊平滑肌中存在ICC起搏细胞,参与起搏胆囊自发节律性电活动和自动节律性收缩活动。
关键词:  胆囊  Cajal间质细胞  动力  起搏
DOI:10.3724/SP.J.1008.2010.0721
投稿时间:2009-06-21修订日期:2010-05-27
基金项目:江西省自然基金(2008GQY0036).
Interstitial cell of cajal regulates motility of gallbladder smooth muscle in guinea pigs
XU Long 1, YU Bao-ping 2*, CHEN Ling-dan2 , L Nong-hua1 , ZHU Xuan 1, CHEN You-xiang1,Yu Baoping
(1.Department of Gastroenterology, the First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi, China; 2.Department of Gastroenterology, People’s Hospital of Wuhan University, Wuhan 430060, Hubei, China)
Abstract:
ObjectiveTo prove that the interstitial cells of cajal (ICCs) are the pacemaker of gallbladder smooth muscle in guinea pigs.MethodsThe contraction of gallbladder was observed by isometric transducer and spontaneous electrophysiology was recorded in vivo and in vitro.ICCs were damaged with methylene and illumination.The ICCs were isolated by collagenase digestion and the morphology of single ICC was analyzed.C-kit expression was detected by laser confocal microscopy to identify the isolated ICCs.Spontaneous currents of ICCs were recorded with EPC-10 Patch Clamp.ResultsSlow wave and action potential were recorded in gallbladder in vivo, with the frequency of slow wave being (58±3.2) bpm (n=16).Methylene blue and illumination treatment led to decrease or disappearance of slow wave and action potentials.Some cells isolated from gallbladder smooth muscle (\[5±1.2\]%, n=12)showed the morphological characteristics of ICCs under light microscope, with (21±4)% of the cells were positive for c-kit.When the membrane potential was at -60 mV, the frequency of spontaneous inward currents was (58.4±3.5) bpm (n=21) and the amplitude was -(72±3.5) pA.ConclusionICC-like cells are present in the gallbladder smooth muscle of the guinea pigs, and participate in the pacemaking of spontaneous current and action potential of gallbladder smooth muscle.
Key words:  gallbladder  interstitial cell of Cajal  motility  pacemaker