Alger Heather M, Rayavarapu Sree, Nagaraju Kanneboyina
Research Center for Genetic Medicine, Children's National Medical Center, Washington, DC, USA.
Methods Enzymol. 2011;489:207-25. doi: 10.1016/B978-0-12-385116-1.00012-1.
Evidence suggests that both immune (cell-mediated and humoral) and nonadaptive immune (endoplasmic reticulum (ER) stress and autophagy) mechanisms play a role in muscle fiber damage and dysfunction in idiopathic inflammatory myopathies (IIM). More recently, the ER stress response pathway, the activation of unfolded protein response, and the ER overload response are being studied to understand their contribution in the progression of IIM. A variety of qualitative and quantitative techniques are used to measure the activation of the endoplasmic reticulum stress response in myopathy. Accurately assessing the activation of ER stress response pathway would not only help in the understanding of disease pathogenesis but would also help to assess the response to therapy. Here, we describe common techniques such as western blotting, immunohistochemistry, immunofluorescence, and determination of mRNA levels for the gene of interest to monitor the ER stress in skeletal muscle tissues.
有证据表明，免疫（细胞介导和体液免疫）和非适应性免疫（内质网（ER）应激和自噬）机制在特发性炎性肌病（IIM）的肌纤维损伤和功能障碍中均起作用。最近，内质网应激反应途径、未折叠蛋白反应的激活以及内质网过载反应正在被研究，以了解它们在IIM进展中的作用。多种定性和定量技术被用于测量肌病中内质网应激反应的激活情况。准确评估内质网应激反应途径的激活不仅有助于理解疾病发病机制，还有助于评估治疗反应。在此，我们描述了诸如蛋白质印迹法、免疫组织化学、免疫荧光以及测定感兴趣基因的mRNA水平等常用技术，以监测骨骼肌组织中的内质网应激。
