单细胞拉曼光谱研究游离脂肪酸对NIT-1胰岛β细胞生理的影响

Effect of Free Fatty Acids on NIT-1 Pancreas Islet β Cell Physiological Changes by the Single Raman Spectrum

运用激光镊子拉曼光谱技术探讨高浓度游离脂肪酸对NIT-1胰岛β细胞生理的影响。实验组细胞以含0.25mmol/L棕榈酸培养12,24,48h,平行对照组中不含任何脂肪酸。通过扫描形式收集各组细胞的拉曼光谱后,使用Hoechs33342/PI荧光染色保留活细胞的光谱,并在OriginPro 8.0系统中比较各组平均光谱的差异。结果显示对照组间的拉曼光谱无明显差异;高脂组光谱在780、1124、1092cm-1及1172cm-1发生拉曼位移,且780、1172、855cm-1及873cm-1峰值强度降低。这些变化的光谱峰在分子归属上指认为细胞内DNA和蛋白质。由此推断高浓度游离脂肪诱导NIT-1细胞损伤在光谱表现为细胞内DNA及蛋白质的结构、含量改变,且该变化与细胞的凋亡有着密切联系。在实验中激光光镊拉曼系统对观察细胞早期损伤有着积极作用。

The possible physiological impacts of NIT-1 pancreas islet β cell attributing to high concentration free fatty acids was explored by laser tweezers raman spectroscopic technique.Cells were cultured with 0.25mmol/L palmacid for 12,24,48h respectively,and one group of cells served as control and the cells were incubated without any free fatty acid.Raman were obtained by the mean of scanning,then live cells＇ spectra were kept by fluorescent staining with Hoechs33342/PI.The spectral difference was analyzed by OriginPro 8.0 system.The shift changes of Raman peaks for high-fat group at 780,1124,1092cm-1 and 1172cm-1 were observed and the peaks intensity declined at 780,1172,855cm-1 and 873cm-1,while there was no obvious difference between control groups.These above peaks mainly correspond to intracelluar DNA and protein.The above results suggested high concentration free fatty acids can impair the NIT-1 cell by changing the structure or content of DNA and protein,which highly relate with apoptosis.In this experiment,Raman spectra have the advantage of observing cellular structure changes in early period.