拉曼光谱技术检测川芎嗪对视网膜色素上皮细胞氧化应激损伤的保护作用-《中国医学物理学杂志》

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Title:: Detection of the protective effect of ligustrazine against oxidative stress damages to retinal pigment epithelium cell via Raman spectroscopy

文章编号:: 1005-202X（2021）07-0864-07

作者:: 谢婷1; 陈阳2; 陈文奕1; 赖巧玲1; 曹改改2; 许云超3; 黄焱1; 1.福建医科大学医学技术与工程学院眼视光学系，福建福州 350004； 2.福建医科大学医学技术与工程学院医学检验学系，福建福州 350004； 3.福建师范大学医学光电科学与技术教育部重点实验室，福建福州 350004

Author(s):: XIE Ting1; CHEN Yang2; CHEN Wenyi1; LAI Qiaoling1; CAO Gaigai2; XU Yunchao3; HUANG Yan1; 1. Department of Ophthalmic Optics, School of Medical Technology and Engineering, Fujian Medical University, Fuzhou 350004, China 2. Department of Medical Laboratory Science, School of Medical Technology and Engineering, Fujian Medical University, Fuzhou 350004, China 3. Key Laboratory of Medical Optoelectronic Science and Technology, Ministry of Education, Fujian Normal University, Fuzhou 350004, China

关键词:: 视网膜色素上皮细胞; 拉曼光谱; 活性氧; 川芎嗪; 氧化应激损伤

Keywords:: Keywords: retinal pigment epithelium cell Raman spectroscopy reactive oxygen ligustrazine oxidative stress damage

分类号:: R318；R34

DOI:: DOI:10.3969/j.issn.1005-202X.2021.07.015

文献标志码:: A

摘要:: 目的：年龄相关性黄斑变性（AMD）是致盲的主要原因之一。氧化应激所引起的视网膜色素上皮（RPE）细胞变性在AMD的发病机制中发挥了关键作用。本文利用无创、无需标签、高灵敏度的拉曼光谱探究川芎嗪对氧化损伤的人RPE细胞的保护机制。方法：进行拉曼光谱采集之前，将除对照组外的其余两组RPE细胞（ARPE-19）用200 μmol/mL H2O2预孵育，24 h后在保护组细胞中加入200 μmol/mL川芎嗪。所有干预结束后，用MultiskanGO （Thermo, USA）微板法测定细胞内活性氧自由基的含量，用InVia微拉曼系统对3组细胞进行拉曼光谱分析。结果：活性氧检测显示川芎嗪对H2O2诱导的RPE细胞氧化应激损伤有较好的抑制作用。拉曼光谱结果显示细胞氧化应激损伤主要体现在氨基酸类分子上，而川芎嗪的加入不仅逆转了归属于脂质的谱带810 cm-1和840 cm-1峰强度比值的变化趋势，还使H2O2干预后显著提高的归属于各氨基酸谱带的峰强度值有所下降。与之前激光光镊拉曼光谱结果不同，活性氧自由基没有触发酰胺的脱酰胺反应，反而是川芎嗪的干预使得归属于酰胺的拉曼光谱强度发生改变。结论：通过对人RPE细胞拉曼光谱变化的分析，揭示过氧化氢和抗氧化剂川芎嗪潜在的作用靶点，为更好地探究RPE细胞氧化应激的损伤机制奠定了基础。

Abstract:: Abstract: Objective Age-related macular degeneration (AMD) is one of the main causes of blindness. Retinal pigment epithelium (RPE) cell degeneration induced by oxidative stress plays a key role in the pathogenesis of AMD. The protective mechanism of ligustrazine against oxidative stress damages to human RPE cells is investigated using non-invasive, label-free and highly-sensitive Raman spectroscopy. Methods Before the collection of Raman spectra, RPE cells (ARPE-19) in 2 groups, except control group, were preincubated with 200 μmol/mL H2O2, and 200 μmol/mL ligustrazine was added to the cells in protection group after 24 h. At the end of all interventions, MultiskanGO (Thermo, USA) microplate method was used for determining intracellular reactive oxygen species (ROS) content, and InVia microRaman system for Raman spectrum analysis. Results The ROS levels showed that ligustrazine had a good inhibitory effect on oxidative stress damages induced by H2O2. Raman spectral results demonstrated that the oxidative stress damages to cells were mainly reflected in amino acids. The addition of ligustrazine not only reversed the changing trend of the peak strength ratio of 810 cm-1 and 840 cm-1 assigned to the lipid, but also reduced the peak strength value of each band assigned to amino acids that was significantly increased after H2O2 intervention. Different from the previous laser tweezers Raman spectral results, ROS did not trigger the deamide reaction of amides, but ligustrazine changed the Raman spectral strength assigned to amides. Conclusion Through Raman spectrum analysis on human RPE cells, the potential biomarkers of hydrogen peroxide and ligustrazine are revealed, which laid a foundation for investigating the mechanism of oxidative stress damages to human RPE cells.

《中国医学物理学杂志》[ISSN:1005-202X/CN:44-1351/R]

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