CAJ | 학술논문

目的探讨基于超高效液相色谱-质谱技术(ultra performance liquid chromatographymass spectrometry,UPLC-MS)的模式识别方法应用于SD大鼠弥漫性轴索损伤(diffuse axonal injury,DAI)模型血浆代谢组学研究的可行性.方法采用改良Marmarous法建立SD大鼠DAI模型.将18只SD大鼠按随机数字表法分为空白对照组、假手术对对照组和打击致伤组,每组6只.应用UPLC-MS技术对空白对照组、假手术对照组及打击致伤组大鼠血浆进行检测,所得数据分别采用主成分分析法(principal component analysis,PCA)、偏最小二乘判别分析法(partial least squares discriminant analysis,PLS-DA)和正交信号偏最小二乘判别分析法(orthogonal signal correction-partial least squares discriminant analysis,OSC-PLS-DA)进行模式识别分析,确定打击致伤组与对照组血浆的代谢差异,比较三种模式识别方法的判别能力.根据PLS-DA模型的变量重要性投影(variable importance in the projection,V1P)值初步筛选出可能的代谢标志物.结果打击致伤组与对照组血浆代谢组存在明显差异,但PCA模式识别未能将打击致伤组和对照组明显区分开,而PLS-DA和OSC-PLS-DA模式识别法可显著将其分开.初步确认7个质荷比(分别为243.102 3,227.103 0,146.165 0,59.049 7,145.1197,218.138 9,169.096 9)对应的化合物可能为大鼠DA1候选的潜在生物标志物.结论建立了利用UPLC-MS技术进行DAI大鼠模型早期的血浆代谢组学研究检测,PLS-DA和OSC-PLS-DA模式识别法明显优于PCA模式识别法,初步筛选出大鼠DAI可能的生物标志物,为DAI早期诊断、治疗及法医学鉴定奠定了基础.
목적 탐토기우초고효액상색보-질보기술(ultra performance liquid chromatographymass spectrometry,UPLC-MS)적모식식별방법응용우SD대서미만성축색손상(diffuse axonal injury,DAI)모형혈장대사조학연구적가행성.방법 채용개량Marmarous법건립SD대서DAI모형.장18지SD대서안수궤수자표법분위공백대조조、가수술대대조조화타격치상조,매조6지.응용UPLC-MS기술대공백대조조、가수술대조조급타격치상조대서혈장진행검측,소득수거분별채용주성분분석법(principal component analysis,PCA)、편최소이승판별분석법(partial least squares discriminant analysis,PLS-DA)화정교신호편최소이승판별분석법(orthogonal signal correction-partial least squares discriminant analysis,OSC-PLS-DA)진행모식식별분석,학정타격치상조여대조조혈장적대사차이,비교삼충모식식별방법적판별능력.근거PLS-DA모형적변량중요성투영(variable importance in the projection,V1P)치초보사선출가능적대사표지물.결과 타격치상조여대조조혈장대사조존재명현차이,단PCA모식식별미능장타격치상조화대조조명현구분개,이PLS-DA화OSC-PLS-DA모식식별법가현저장기분개.초보학인7개질하비(분별위243.102 3,227.103 0,146.165 0,59.049 7,145.1197,218.138 9,169.096 9)대응적화합물가능위대서DA1후선적잠재생물표지물.결론 건립료이용UPLC-MS기술진행DAI대서모형조기적혈장대사조학연구검측,PLS-DA화OSC-PLS-DA모식식별법명현우우PCA모식식별법,초보사선출대서DAI가능적생물표지물,위DAI조기진단、치료급법의학감정전정료기출.
Objective To investigate the feasibility in identification of plasma metabolomics of SD rats with diffuse axonal injury (DAI) in early phase using the ultra performance liquid chromatography -mass spectrometry (UPLC-MS)-based pattern recognition method.Methods Models of DAI in SD rats were induced by the modified Marmarous method.Eighteen SD rats were allocated to blank control group,sham operation group and injury group according to the random number table,with 6 rats per group.UPLC-MS technique was applied for examination of plasma samples from each groups.Pattern-recognition techniques of principal component analysis (PCA),partial least squares discriminant analysis (PLS-DA) and orthogonal signal correction-partial least squares discriminant analysis (OSC-PLS-DA) were involved to analyze the plasma metabolites and identify the differences among these groups.Variable importance in projection (VIP) of PLS-DA models was involved to screen potential metabolic markers.Results Plasma metabolomics differed significantly among the three groups.PCA pattern recognition model failed to tell apart the difference,but PLS-DA and OSC-PLS-DA identified distinct differences among these three groups.Seven compounds (mass to charge ratios were 243.102 3,227.103 0,146.165 0,59.049 7,145.119 7,218.138 9 and 169.096 9 respectively) were initially identified as potential candidate biomarkers of DAI in SD rats.Conclusions Early detection of plasma metabolomics of SD rats with DAI has been constructed using the technique of UPLC-MS,but PLS-DA and OSC-PLS-DA are superior to PCA in pattern recognition.UPLC-MS technique helps to examine the potential biomarkers of DAI in SD rats,which lays the foundation for research of early diagnosis,treatment and forensic identification related to DAI.