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Company: Eppendorf
Catalog#: 3111000.165
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Double Labeling of PDGFR-β and α-SMA in Swine Models of Acute Kidney Injury to Detect Pericyte-to-Myofibroblast Transdifferentation as Early Marker of Fibrosis
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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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