【摘要】 研究细胞内Ca2+对低温连续灌流 120 h的离体肾的影响.对经连续灌流的离体肾脏组织进行冰冻切片处理,切片经Fluo 3/AM荧光剂染色后分成三组.Ⅰ组利用无 Ca2+ 孵育液,Ⅱ组利用添加普鲁卡因的无 Ca2+ 孵育液,Ⅲ组利用添加异搏定的无Ca2+孵育液,分别孵育30 min,并采用激光扫描共聚焦显微镜(LSCM)检测肾小管上皮细胞亚结构中Ca2+浓度的变化.结果表明: 低温适应过程中肾小管上皮细胞内Ca2+浓度升高主要源于内钙变化,持续时间不长;被普鲁卡因选择性阻断的内质网膜Ryanodine受体系统介导的Ca2+释放可能为该过程中Ca2+的来源.同时Ca2+检测手段对实验过程中Ca2+浓度变化的湮灭效应应该加以注意.更多还原

【Abstract】 The influence of calcium on the surviving kidney perfused continuously for 120 h at low temperature was studied. The off-body cat kidney was perfused with calcium-free perfusate, then frozen sections of the kidney specimen were prepared and divided into 3 groups, all treated with Fluo-3/AM. Each group was incubated for 30 min, the 1st group with calcium-free perfusate, the 2nd with calcium-free perfusate plus verapamil and the 3rd with calcium-free perfusate plus procaine. Finally the observation of calcium fluorescence intensity was conducted in the proximal tubular epithelial cells by the laser scanning confocal microscopy (LSCM).The results lead to two conclusions: during the cold-adaptation period the rise of the calcium concentration mainly comes from calcium pools in the cell; and the release induced by the Ryanodine-receptor system of the calcium pools in the endoplasmic reticulum may be the source. The influence brought by the method in this experiment also cannot be overlooked.更多还原