光谱学与光谱分析, 2020, 40 (9): 2751, 网络出版: 2020-11-25  

对KOA模型大鼠血清、 膝关节肌肉和滑膜组织的SERS分析

Surface-Enhanced Raman Spectroscopy Analysis on the Serum, Muscular and Synovial Tissue of the Knee in Knee Osteoarthritis Model Rats
作者单位
1 安徽中医药大学研究生院, 安徽 合肥 230012
2 中国科学院合肥物质科学研究院, 安徽 合肥 230031
3 安徽中医药大学针灸推拿学院, 安徽 合肥 230012
4 安徽省中医药科学院针灸经络研究所, 安徽 合肥 230038
摘要
通过比较分析正常大鼠与膝骨关节炎模型(KOA)大鼠血清、 膝关节肌肉和滑膜组织的表面增强拉曼光谱(SERS), 为KOA生物学改变提供实验基础。 同条件下饲养普通级健康雄性SD大鼠20只, 随机分为正常对照组(简称“正常组”)和KOA模型组(简称“模型组”), 每组10只。 采用左膝关节腔内注射0.03 mol·L-1的L-半胱氨酸与4%木瓜蛋白酶混合物方法制备KOA模型, 并于复制成功4周后取材。 采用银纳米基底液检测大鼠血清和膝关节肌肉、 滑膜组织中的表面增强拉曼谱峰, 应用NGLabSpec软件比较两组拉曼频移和特征峰的差异, 应用OriginPro 8.5软件分析拉曼光谱图。 结果: 在血清中, 拉曼频移400~2 000 cm-1区间内, 正常组特征峰有12个, 模型组有14个, 且模型组大部分特征峰强度低于正常组, 两组在495, 883和1 447 cm-1等处出现较为显著的差异性特征峰; 在膝关节肌肉组织中, 正常组特征峰有12个, 模型组有13个, 二者的同质性特征峰的拉曼强度存在显著差异, 模型组以950和1 237 cm-1为代表的多处同质性特征峰的峰强显著升高; 在滑膜组织中, 正常组特征峰有10个, 模型组有15个, 两组共性特征峰的峰强变化多不明显, 但在655, 950, 1 335和1 447 cm-1处的同质性特征峰表现出峰强的明显差异, 在655和950 cm-1峰为模型组显著升高, 而1 335和1 447 cm-1两峰相对强度为模型组显著降低。 结果表明: KOA模型导致血清、 膝关节肌肉和滑膜组织的同质性特征峰数量显著减少, 差异性物质增多, 物质代谢平衡被严重打破, SERS是一种快速准确的检测方法, 可以用于KOA模型的检测。
Abstract
Surface-enhanced Raman Spectorscopy (SERS) spectra of serum, knee muscles and synovial tissues of normal rats and Model of Knee Osteoarthritis (KOA) are analyzed, providing an experimental basis for biological changes of KOA. Twenty normal healthy male SD rats were raised under the same conditions and randomly divided into normal control group (referred to as “normal group”) and KOA model group (referred to as “model group”), with 10 rats in each group. The KOA model was prepared by intra-articular injection of 0.03 mol·L-1 of l-cysteine and 4% papain. The classification of surficial enhanced Raman spectra in serum, knee muscle and synovial tissue of rats was detected by silver nanometer substrate fluid. NGLabSpec software was used to compare the differences of Raman frequency shift and characteristic peaks between the two groups, and OriginPro 8.5 software was used to analyze Raman spectra. In the serum, within the range of Raman frequency shift 400~2 000 cm-1, there were 12 characteristic peaks in the normal group and 14 in the model group, and the peak strength of most characteristic peaks in the model group was lower than that in the normal group, and significant difference characteristic peaks appeared in 495, 883 and 1 447 cm-1 of the two groups; in the muscle tissue of the knee joint, there were 12 characteristic peaks in the normal group and 13 in the model group, there were significant differences in the Raman strength of the characteristic peaks of homogeneity between the two groups; in synovial tissues and the normal group has 10 characteristic peaks, has 15 model group, two groups of the common characteristic peak of peak intensity change is not obvious, but in the 655, 950, 1 335, 1 447 cm-1 of the homogeneous characteristic peaks show that the peak strength of significant differences, in 655 and 950 cm-1 of a significant rise in the model group, and 1 335 and 1 447 cm-1 model is the relative strength of two groups decreased significantly. KOA model led to a significant decrease in the number of homogenous characteristic peaks of serum, knee muscle and synovial tissue, and an increase in different substances. Thus the metabolic balance of substances was seriously broken. SERS is a fast and accurate detection method, which can be used for the detection of KOA model.

王璐璐, 刘磊, 李盼, 王洁, 何璐, 吴子建, 杨良保, 胡玲. 对KOA模型大鼠血清、 膝关节肌肉和滑膜组织的SERS分析[J]. 光谱学与光谱分析, 2020, 40(9): 2751. WANG Lu-lu, LIU Lei, LI Pan, WANG Jie, HE Lu, WU Zi-jian, YANG Liang-bao, HU Ling. Surface-Enhanced Raman Spectroscopy Analysis on the Serum, Muscular and Synovial Tissue of the Knee in Knee Osteoarthritis Model Rats[J]. Spectroscopy and Spectral Analysis, 2020, 40(9): 2751.

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