School of Biomedical Sciences, The University of Queensland, Brisbane, Queensland, Australia.
PLoS One. 2013;8(4):e60255. doi: 10.1371/journal.pone.0060255. Epub 2013 Apr 2.
The role of the free fatty acid (FFA) receptor and the intracellular metabolites of linoleic acid (LA) in LA-stimulated increase in cytosolic free calcium concentration ([Ca(2+)]i) was investigated. [Ca(2+)]i was measured using Fura-2 as indicator in rat pancreatic β-cells in primary culture. LA (20 µM for 2 min) stimulated a transient peak increase followed by a minor plateau increase in [Ca(2+)]i. Elongation of LA stimulation up to 10 min induced a strong and long-lasting elevation in [Ca(2+)]i. Activation of FFA receptors by the non-metabolic agonist GW9508 (40 µM for 10 min) resulted in an increase in [Ca(2+)]i similar to that of 2-min LA treatment. Inhibition of acyl-CoA synthetases by Triacsin C suppressed the strong and long-lasting increase in [Ca(2+)]i. The increase in [Ca(2+)]i induced by 2 min LA or GW9508 were fully eliminated by exhaustion of endoplasmic reticulum (ER) Ca(2+) stores or by inhibition of phospholipase C (PLC). Removal of extracellular Ca(2+) did not influence the transient peak increase in [Ca(2+)]i stimulated by 2 min LA or GW9508. The strong and long-lasting increase in [Ca(2+)]i induced by 10 min LA was only partially suppressed by extracellular Ca(2+) removal or thapsigargin pretreatment, whereas remaining elevation in [Ca(2+)]i was eliminated after exhaustion of mitochondrial Ca(2+) using triphenyltin. In conclusion, LA stimulates Ca(2+) release from ER through activation of the FFA receptor coupled to PLC and mobilizes mitochondrial Ca(2+) by intracellular metabolites in β-cells.
研究了游离脂肪酸(FFA)受体和亚油酸(LA)的细胞内代谢物在 LA 刺激细胞浆游离钙浓度([Ca(2+)]i)增加中的作用。在原代培养的大鼠胰岛β细胞中,使用 Fura-2 作为指示剂测量[Ca(2+)]i。LA(20 μM,2 分钟)刺激[Ca(2+)]i 产生短暂的峰增加,随后是较小的平台增加。将 LA 刺激延长至 10 分钟会导致[Ca(2+)]i 强烈且持久的升高。非代谢激动剂 GW9508(40 μM,10 分钟)激活 FFA 受体导致[Ca(2+)]i 增加与 2 分钟 LA 处理相似。三碘乙酸酯 C 抑制酰基辅酶 A 合成酶抑制强烈且持久的[Ca(2+)]i 增加。2 分钟 LA 或 GW9508 诱导的[Ca(2+)]i 增加完全被内质网(ER)钙库耗竭或磷脂酶 C(PLC)抑制消除。去除细胞外 Ca(2+)不会影响 2 分钟 LA 或 GW9508 刺激的[Ca(2+)]i 短暂峰值增加。10 分钟 LA 诱导的[Ca(2+)]i 强烈且持久的增加仅部分被细胞外 Ca(2+)去除或 thapsigargin 预处理抑制,而使用三苯基锡耗尽线粒体 Ca(2+)后,剩余的[Ca(2+)]i 消除。总之,LA 通过激活与 PLC 偶联的 FFA 受体刺激 ER 中的 Ca(2+)释放,并在β细胞中通过细胞内代谢物动员线粒体 Ca(2+)。