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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 effects of high concentration of free fatty acids on the islet β cell physiology of NIT-1 were investigated by laser tweezers Raman spectroscopy. The cells in the experimental group were incubated with 0.25 mmol / L palmitic acid for 12, 24 and 48 h, and the parallel control group did not contain any fatty acid. After the Raman spectra of each group of cells were collected by scanning, the spectrum of viable cells was preserved using Hoechs 33342 / PI fluorescence staining and the difference between the mean spectra for each group was compared in the OriginPro 8.0 system. The results showed that there was no significant difference in the Raman spectra between the control groups. Raman shift was observed at 780, 1124, 1092 cm-1 and 1172 cm-1 in the hyperlipidemic group, and the peak intensities at 780, 1128, 855 cm-1 and 873 cm-1 were decreased. These changing spectral peaks are referred to as intracellular DNA and proteins at the molecular attribution. It was concluded that high concentrations of free fat-induced NIT-1 cell damage in the spectrum of DNA and protein showed the structure and content of cells changed, and the change is closely related with cell apoptosis. Laser tweezers Raman system in the experiment to observe the early cell damage has a positive effect.