Ca2+-钙调蛋白在M3R介导逼尿肌细胞收缩中的作用

摘要

背景:毒蕈碱受体在调节逼尿肌细胞收缩过程中起重要作用,其受体亚型M3R直接介导逼尿肌细胞收缩.Ca2+是刺激逼尿肌细胞收缩的直接因素,Ca2+有数十种受体结合蛋白质,Ca2+通过与不同受体蛋白结合调节不同反应.目的:探讨Ca2+-钙调蛋白在M3R介导逼尿肌细胞收缩中的作用.设计:以逼尿肌细胞为观察对象,对比观察.单位:解放军第三军医大学西南医院泌尿中心.材料:实验在重庆西南医院中心实验室完成,实验动物选择健康雌性Wistar大鼠.方法:将原代培养逼尿肌细胞分为实验组和对照组,接种于6孔培养板上培养,实验组培养细胞70%融合时加入1×10-4 mmol/L卡巴胆碱和毒蕈碱受体亚型M2R拮抗剂,分别阻断M3R、M2R,采用Ca2+浓度和钙调蛋白活性检测试剂盒分别测定两组逼尿肌细胞Ca2+浓度和钙调蛋白活性.主要观察指标:两组细胞内[Ca2+]i浓度、钙调蛋白活性变化.结果:实验组的[Ca2+]i和钙调蛋白的平均通道荧光对数值高于对照组(3.26±0.38,2.06±0.12,P＜0.01);(2.87±0.34,2.14±0.24,P＜0.05).结论:实验结果提示Ca2+-钙调蛋白以信号传导途径参与了M3R介导逼尿肌细胞收缩的调节过程.%BACKGROUND: Muscarine receptor plays a key role in adjusting contraction of detrusor muscle cell, and M3R, isoforms of its receptor, can mediate contraction of detrusor muscle cell directly. Ca2+ is the direct factor in stimulating contraction of detrusor muscle cell. Of several 10 kinds of Ca2+conjugable receptors protein, Ca2+ conjugated with different receptor proteins can adjust various reactions.OBJECTIVE: To investigate the effect of Ca2+-calmodulin (Ca2+-CaM) on contraction of M3R-mediated detrusor muscle cell.DESIGN: Compared observation .on the basis of detrusor muscle cell.SETTING: Ourological Center of Southwest Hospital of the Third Military Medical University of Chinese PLA.MATERIALS: The experiment was completed at Central Laboratory of Southwest Hospital of the Third Military Medical University of Chinese PLA. Healthy Wistar rats were selected in this study.METHODS: The primary cultured detrusor muscle cells were divided into experimental group and control group. Cells were inoculated in 6-well plate, and 10-4 mmol/L carbachol and M2R antagonist were added to cells of the experimental group during 70% confluence to block M3R and M2R respectively. Ca2+ concentration and CaM activity were detected by Ca2+ test kit and CaM test kit respectively.MAIN OUTCOME MEASURES: Changes of [Ca2+]I concentration and CaM activity of cells in both groups.RESULTS: The mean channel fluorescence values (log) of [Ca2+]I and CaM were higher in the experimental group than those in the control group(3.26±0.38, 2.06±0.12; 2.87±0.34, 2.14±0.24, P ＜ 0.05).CONCLUSION: Results in this study suggest that Ca2+-CaM participates in adjusting contraction of M3R-mediated detrusor muscle cells through signal transduction.