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Author:
Date:
2020-10-05
[Abstract] Growing evidences suggest that peritubular capillaries pericytes are the main source of scar-forming myofibroblasts during chronic kidney disease (CKD), as well as early phases of acute kidney injury (AKI). In a swine model of sepsis and I/R (Ischemia Reperfusion) injury-induced AKI we demonstrated that renal pericytes are able to transdifferentiate toward α-SMA+ myofibroblasts leading to interstitial fibrosis. Even if precise pericytes identification requires transmission electron microscopy and the co-immunostaining of several markers (i.e., Gli, NG2 chondroitin sulphate proteoglycan, CD146, desmin or CD73) and emerging new markers (CD248 or TEM1, endosialin), previous studies suggested that PDGFR-β could be used as marker for renal pericytes characterization. ...
[摘要] [摘要]越来越多的证据表明，肾小管周围的毛细血管周细胞是慢性肾脏病（CKD）以及急性肾损伤（AKI）早期形成疤痕的成纤维细胞的主要来源。在败血症和I / R（缺血再灌注）损伤诱导的AKI的猪模型中，我们证明了肾周细胞能够向α- SMA +肌成纤维细胞转分化，从而导致间质纤维化。即使精确周细胞识别需要透射电子显微镜和几个标志物联合免疫（即。，的Gli ，NG2硫酸软骨素蛋白聚糖，CD146，结蛋白或CD73）和新兴的新的标志物（CD248或TEM1，唾液酸蛋白），以往的研究表明，PDGFR-β可用作肾周细胞表征的标志物。最近，对PDGFR-β和α-SMA进行了双重免疫荧光染色，以鉴定在纤维化发展的早期受损激活的周细胞（PDGFR-β + /α-SMA +细胞）。我们的数据强调了肾周细胞在败血症和I / R相关性AKI的生理病理中的关键作用。在该协议中，我们描述了猪福尔马林固定石蜡包埋（FFPE）肾脏活检中PDGFR-β和α-SMA双重免疫荧光染色的程序以及图像分析和定量方法。
[背景】肾脏纤维化被认为是主要负责肾脏疾病的进展，其与肾的损伤后的容量有限，再生有关。进行性肾脏疾病中间质纤维化的主要来源（Simone等人，2014 ; ...
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Author:
Date:
2020-08-05
[Abstract] In order for the brain to function properly, a carefully orchestrated homeostasis must be maintained. To help regulate this delicate balance, the brain has developed a highly selective blood-brain barrier (BBB). Under normal conditions, the BBB excludes harmful blood-borne material from the brain parenchyma. However, numerous neuropathological conditions can disrupt this barrier, causing BBB permeability and subsequent CNS dysfunction. Understanding the mechanisms involved in BBB permeability are essential to elucidating the pathology of various neurological disorders as well as identifying methods for drug delivery to the CNS. Here, we describe several in vivo methods to measure BBB permeability in mice using an array of diverse sized tracers including exogenous 376 Da ...
[摘要] [摘要]为了使大脑正常工作，必须维持精心安排的体内平衡。为了帮助调节这种微妙的平衡，大脑已经形成了一种高度选择性的血脑屏障（BBB）。在正常情况下，血脑屏障将有害的血液传播物质排除在脑实质之外。然而，许多神经病理学条件可以破坏这一屏障，导致BBB通透性和随后的CNS功能障碍。了解BBB通透性的机制对于阐明各种神经系统疾病的病理学以及确定药物输送到中枢神经系统的方法至关重要。在这里，我们描述了几种在体内使用不同大小的示踪剂（包括外源性376da荧光素盐、66.5kDa牛血清白蛋白、70kDa右旋糖酐以及内源性160kDa小鼠IgG）来测量BBB在小鼠体内的通透性。当静脉注射时，这些物质会被BBB排除在健康大脑之外。然而，BBB功能障碍可使这些示踪剂进入大脑，这种积聚可通过分光光度法、荧光显微镜和免疫组织化学进行测量。我们还描述了一种利用产气荚膜梭菌epsilon毒素诱导BBB通透性的方法。最后，我们将简要讨论每种方法的优缺点及其适当的下游应用。
[背景] ...
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