Verzola Daniela, Milanesi Samantha, Costigliolo Francesca, Garibotto Giacomo
Clinica Nefrologica, Dialisi, Trapianto, Università di Genova e Ospedale Policlinico San Martino, Genoa, Italy.
Bio Protoc. 2019 Jan 20;9(2):e3144. doi: 10.21769/BioProtoc.3144.
Growing evidence suggests the involvement of TLR4, a receptor in the innate immune system, in muscle loss in uremia. Recently, we have evaluated TLR4 in human skeletal muscle from chronic kidney disease patients, by immunohistochemistry and image analysis. Unlike the commonly-used Western blot method, immunohistochemistry allows for the observation of protein distribution in the intact tissue while, image analysis, its quantification. In fact, our data highlighted our hypothesis that an enhanced TLR4 skeletal muscle cell expression contributes to the activation of the downward inflammatory pathway in uremic sarcopenia. In this protocol, we describe the procedure for immunostaining TLR4 in human skeletal muscle and for quantifying it by image analysis.
越来越多的证据表明，先天免疫系统中的一种受体——Toll样受体4（TLR4）参与了尿毒症患者的肌肉流失。最近，我们通过免疫组织化学和图像分析，对慢性肾病患者的人体骨骼肌中的TLR4进行了评估。与常用的蛋白质印迹法不同，免疫组织化学能够观察完整组织中的蛋白质分布，而图像分析则可对其进行定量分析。事实上，我们的数据突出了我们的假设，即TLR4在骨骼肌细胞中表达增强，有助于激活尿毒症性肌肉减少症中的下行炎症途径。在本方案中，我们描述了在人体骨骼肌中对TLR4进行免疫染色并通过图像分析对其进行定量的程序。
