王莉, 付荣国, 刘晓丹, 桂保松, 孙强, 陈晨, 赵玉峰, 董蕾
西安交通大学医学院第二附属医院，西安 710004；西安交通大学医学院，西安 710061；第四军医大学教学实验中心，西安 710032
为明确亚油酸影响胰岛 细胞的胞浆游离钙水平([Ca2+]i)的作用和机制，本研究运用胶原酶灌注消化法原代培养大 鼠胰岛细胞，使用钙离子荧光指示剂Fluo-3 负载细胞后在共聚焦显微镜下观察Fluo-3 荧光强度以反映[Ca2+]i 变化，灌流给 药观察亚油酸等药物的作用，记录结束后采用免疫细胞化学染色方法鉴定 细胞。结果显示，亚油酸(20 mol/L)刺激 细 胞 [Ca2+]i 升高，表现为最初的峰型升高和随后的较为持久的平台期升高。平台期升高可被去除细胞外液中Ca2+ 所阻断，也 可被瞬时受体电位(transient receptor potential, TRP)通道的阻断剂La3+ 所抑制，峰型升高和平台期升高均被耗竭细胞内钙库 的Ca2+ 或者抑制磷脂酶C 活性所阻断。结果表明，亚油酸通过刺激 细胞内钙库的Ca2+ 释放和激活TRP 通道，导致[Ca2+]i 升高。
[Effects of linoleic acid on intracellular calcium concentration in primarily cultured rat pancreatic β -cells and underlying mechanism.] [Ariticle in Chinese]
WANG Li, FU Rong-Guo, LIU Xiao-Dan, GUI Bao Song, SUN Qiang, CHEN Chen, ZHAO Yu-Feng, DONG Lei
The Second Affiliated Hospital of Medical School, Xi’an Jiao Tong University, Xi’an 710004, China； Medical School of Xi’an Jiao Tong University, Xi’an 710061, China； Experiment Teaching Center, Fourth Military Medical University, Xi’an 710032, China
In this study, we investigated the mechanism of linoleic acid-stimulated increase in intracellular calcium concentration ([Ca2+]i) in pancreatic islet β-cells. Pancreatic islet cells were primarily isolated from rats and cultured for the experiments. The cells were loaded with Fluo-3/AM, the indicator of [Ca2+]i, and the intensity of Fluo-3 was measured using confocal microscope. The islet β-cells were identified by immunocytochemical staining with insulin antibody after recording. The drugs were given by perfusion system. The results showed that linoleic acid (20 μmol/L) stimulated [Ca2+]i increase with the first peak increase and the following plateau increase. Linoleic acid-stimulated [Ca2+]i increase was partly inhibited by removal of extracellular calcium and by transient receptor potential (TRP) channel blocker, La3+, and it was totally blocked by exhaustion of intracellular calcium stores and inhibition of phospholipase C. It is concluded that linoleic acid stimulates [Ca2+]i increase in islet β-cells through both extracellular calcium influx via TRP channels and calcium release from intracellular calcium stores.
王莉, 付荣国, 刘晓丹, 桂保松, 孙强, 陈晨, 赵玉峰, 董蕾. 亚油酸刺激胰岛β 细胞胞浆游离钙水平升高的机制[J]. 生理学报 2010; 62 (6): 529-534.
WANG Li, FU Rong-Guo, LIU Xiao-Dan, GUI Bao Song, SUN Qiang, CHEN Chen, ZHAO Yu-Feng, DONG Lei. [Effects of linoleic acid on intracellular calcium concentration in primarily cultured rat pancreatic β -cells and underlying mechanism.] [Ariticle in Chinese] . Acta Physiol Sin 2010; 62 (6): 529-534 (in Chinese with English abstract).