Department of Integrative Medical Biology, Anatomy, Umeå University, S-90187 Umeå, Sweden.
BMC Musculoskelet Disord. 2011 Aug 10;12:181. doi: 10.1186/1471-2474-12-181.
The trapezius muscle is a neck muscle that is susceptible to chronic pain conditions associated with repetitive tasks, commonly referred to as chronic work-related myalgia, hence making the trapezius a muscle of clinical interest. To provide a basis for further investigations of the proteomic traits of the trapezius muscle in disease, two-dimensional difference gel electrophoresis (2D-DIGE) was performed on the healthy trapezius using vastus lateralis as a reference. To obtain as much information as possible from the vast proteomic data set, both one-way ANOVA, with and without false discovery rate (FDR) correlation, and partial least square projection to latent structures with discriminant analysis (PLS-DA) were combined to compare the outcome of the analysis.
The trapezius and vastus lateralis showed significant differences in metabolic, contractile and regulatory proteins, with different results depending on choice of statistical approach and pre-processing technique. Using the standard method, FDR correlated one-way ANOVA, 42 protein spots differed significantly in abundance between the two muscles. Complementary analysis using immunohistochemistry and western blot confirmed the results from the 2D-DIGE analysis.
The proteomic approach used in the present study combining 2D-DIGE and multivariate modelling provided a more comprehensive comparison of the protein profiles of the human trapezius and vastus lateralis muscle, than previously possible to obtain with immunohistochemistry or SDS-PAGE alone. Although 2D-DIGE has inherent limitations it is particularly useful to comprehensively screen for important structural and metabolic proteins, and appears to be a promising tool for future studies of patients suffering from chronic work related myalgia or other muscle diseases.
斜方肌是颈部肌肉,易患与重复任务相关的慢性疼痛疾病,通常称为慢性与工作相关的肌痛,因此斜方肌是一种具有临床意义的肌肉。为了为进一步研究斜方肌疾病的蛋白质组学特征提供基础,使用股外侧肌作为参考,对健康的斜方肌进行二维差异凝胶电泳(2D-DIGE)。为了从大量的蛋白质组数据集中尽可能多地获取信息,结合单向方差分析(ANOVA),包括有和无错误发现率(FDR)相关性的分析,以及偏最小二乘投影到潜在结构判别分析(PLS-DA),用于比较分析结果。
斜方肌和股外侧肌在代谢、收缩和调节蛋白方面存在显著差异,不同的结果取决于统计方法和预处理技术的选择。使用标准方法,FDR 相关的单因素方差分析,两种肌肉之间有 42 个蛋白点的丰度有显著差异。使用免疫组织化学和 Western blot 进行补充分析,证实了 2D-DIGE 分析的结果。
本研究中使用的 2D-DIGE 和多元建模相结合的蛋白质组学方法,比以前使用免疫组织化学或 SDS-PAGE 单独进行的方法,更全面地比较了人类斜方肌和股外侧肌的蛋白质图谱。尽管 2D-DIGE 存在固有局限性,但它特别适用于全面筛选重要的结构和代谢蛋白,并且似乎是未来研究慢性与工作相关的肌痛或其他肌肉疾病患者的有前途的工具。