论文部分内容阅读
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1×10-4mmol/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: Muscarinic receptors play an important role in the regulation of detrusor contraction. Its receptor subtype M3R directly mediates detrusor myocyte contraction. Ca2 + is a direct factor to stimulate detrusor cell contraction. Ca2 + has dozens of receptor-binding proteins. Ca2 + regulates different responses by binding to different receptor proteins. AIM: To investigate the role of Ca2 + - calmodulin in M3R-mediated contraction of detrusor myocytes. Design: detrusor cells as the observation object, comparative observation. Unit: Southwest Hospital, Third Military Medical University, Urology Center. MATERIALS: Experiments were performed in the Central Laboratory of Southwest Hospital of Chongqing, and healthy female Wistar rats were selected as experimental animals. Methods: Primary cultured detrusor myocytes were divided into experimental group and control group and inoculated into 6-well culture plates. When 70% of cultured cells in the experimental group were fused, 1 × 10-4 mmol / L carbachol and muscarinic Receptor subtype M2R antagonist, respectively, blocking the M3R, M2R, using Ca2 + concentration and calmodulin activity detection kit were measured detrusor cells Ca2 + concentration and calmodulin activity. MAIN OUTCOME MEASURES: Changes in [Ca2 +] i concentration and calmodulin activity in both groups. Results: The mean channel fluorescence logarithm of [Ca2 +] i and calmodulin in the experimental group was significantly higher than that in the control group (3.26 ± 0.38,2.06 ± 0.12, P <0.01); (2.87 ± 0.34, 2.14 ± 0.24, P <0.05) . Conclusion: The results suggest that Ca2 + - calmodulin is involved in the regulation of the contraction of detrusor cells by M3R signaling pathway.