一种动态观察大鼠胰腺腺泡细胞Ca^(2+)浓度的流式检测方法

A method of dynamic monitoring of cytosolic calcium concentration of rat pancreatic acinar cells

目的探索一种简便的动态检测胰腺腺泡细胞内钙离子浓度的方法。方法无菌采集大鼠胰腺,去除脂肪和淋巴组织,37℃预热的胶原酶消化胰腺组织,分离大鼠胰腺腺泡细胞。加入钙离子荧光探针Fluo-3/AM负载腺泡细胞,流式细胞仪动态监测腺泡细胞内钙离子浓度变化。结果对照组腺泡细胞处于静息状态,钙离子荧光强度稳定,8只大鼠胰腺腺泡细胞内钙离子浓度随时间变化趋势一致,平均荧光强度值为307.75±36.45(n=8);刺激组加入牛磺胆酸钠后腺泡细胞钙离子荧光强度升高,第27.76秒达峰值,检测结果稳定。结论流式细胞仪为腺泡细胞钙离子浓度和细胞功能研究、及后续急性胰腺炎发病机制的研究提供另一简便易行的方法。

Objective To explore a convenient method for dynamic monitoring of cytosolic calcium concentration of rat pancreatic acinar cells.Methods The rat pancreas was collected aseptically and the fat and lymph tissues were removed.Then the rat pancreatic acinar cells were isolated by collagenase preheated at 37℃.The pancreatic acinar cells were incubated with Fluo-3/AM.The dynamic changes of calcium concentration were real-time monitored by flow cytometry.Results The acinar cells of the control group were in a static status,there was no change in the cell fluorescence intensity over time.The calcium concentration in the acinar cells of the 8 rats in this experiment showed a consistent trend over time,with an average fluorescence intensity value of 307.75±36.45(n=8).The cell fluorescence intensity increased and reached the peak value in 27.76 seconds when adding the irritant sodium taurocholate to the cells compared with that before the stimulation.Conclusion Flow cytometry provides another convenient method for the study of calcium cocentration and cell function of acinar cells and the subsequent study of the pathogenesis of acute pancreatitis.