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目的:利用荧光标记的长链脂肪酸,建立一种用于评估动物外周组织游离脂肪酸摄取能力的新方法,为研究体内脂肪酸代谢提供新途径。方法:小鼠腹腔注射和尾静脉注射氟化硼二吡咯荧光标记脂肪酸类似物(BODIPY FL C16),荧光显微镜下观察肝脏和脂肪组织荧光强度。对肝脏和脂肪组织进行脂质抽提,并用酶标仪检测血清、肝脏和脂肪组织相对荧光强度。结果:腹腔和尾静脉注射BODIPY FL C16后均能在小鼠血清[(32 096±2 053)RFU;(22 933±2 430)RFU],肝脏[(40.90±5.08)RFU/mg;(25.70±6.53)RFU/mg]和脂肪组织[(576±90)RFU/mg;(4.990±0.586)RFU/mg]检测到增强的荧光信号(P=0.000);腹腔注射BODIPY FL C16的浓度与外周组织的荧光强度呈正相关(P=0.000);腹腔注射BODIPY FL C16在30~100 min能够维持一个相对脂肪酸代谢的稳态。结论:成功建立了一种体内研究荧光标记脂肪酸摄取的新方法,该方法具有稳定性、重现性,能够准确地反映外周组织脂肪酸摄取情况。
OBJECTIVE: To establish a new method for assessing the uptake of free fatty acids in the peripheral tissues of animals using fluorescently labeled long chain fatty acids, and to provide a new approach for the study of fatty acid metabolism in vivo. Methods: BODIPY FL C16 was injected intraperitoneally into the tail vein and injected into the caudal vein. The fluorescence intensity of the liver and adipose tissue was observed under a fluorescence microscope. Lipids were extracted from liver and adipose tissue, and the relative fluorescence intensity of serum, liver and adipose tissue were detected by microplate reader. RESULTS: After intraperitoneal and caudal vein injection of BODIPY FL C16, mice in the serum ((32 096 ± 2 053) RFU; (22 933 ± 2 430) RFU] and liver [(40.90 ± 5.08) RFU / mg; (P = 0.000) ± 6.53) RFU / mg] and adipose tissue [(576 ± 90) RFU / mg; (4.990 ± 0.586) RFU / mg] (P = 0.000). Intraperitoneal injection of BODIPY FL C16 maintained a relative steady state of fatty acid metabolism at 30-100 min. CONCLUSION: A novel method for the study of uptake of fluorescently labeled fatty acids in vivo has been successfully established. The method is stable and reproducible and accurately reflects the uptake of fatty acids in peripheral tissues.