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基于探针的共聚焦激光诱导内镜在猪食管和胃黏膜中对表皮生长因子受体和生存素的分子成像:概念验证。

Molecular imaging of epidermal growth factor-receptor and survivin in vivo in porcine esophageal and gastric mucosae using probe-based confocal laser-induced endomicroscopy: proof of concept.

机构信息

Division of Gastroenterology and Hepatology, H.H. Chao Comprehensive Digestive Disease Center, University of California, Irvine, School of Medicine, CA, USA.

出版信息

J Physiol Pharmacol. 2012 Jun;63(3):303-7.

Abstract

UNLABELLED

Confocal laser-induced endomicroscopy (CLE) enables in vivo, real time visualization of the subsurface cells and tissue structures in gastrointestinal mucosa at a subcellular resolution of ≈1000x magnification. The aims of this pilot study were to establish a principle of molecular imaging and determine in vivo expression of epidermal growth factor receptor (EGF-R) and survivin in porcine esophageal and gastric mucosa using probe-based CLE (pCLE) and topically applied FITC-labeled antibodies. Studies were performed in anesthetized pigs. During endoscopy FITC-labeled antibodies against EGF-R and survivin were either sprayed onto esophageal and gastric mucosa in preselected areas or administered via submucosal injection. Thirty minutes later pCLE was performed using a through-the-scope probe (GastroFlex UHD, Cellvizio, Mauna Kea Technologies, Paris, France) to determine cellular and tissue localization of EGF-R and survivin. Then the pigs were euthanized and esophageal and gastric walls from the areas sprayed or injected with antibodies were collected for histologic examination under epifluorescence microscopy.

RESULTS

CLE enabled visualization of EGF-R and survivin in esophageal and gastric mucosa and this was confirmed by histology. In the esophagus both EGF-R and survivin were localized predominantly to the keratinocyte progenitor cells. In the stomach, EGF-R was localized to progenitor zone cells and some epithelial cells. Localization of survivin was similar, but involved more surface epithelial cells. This study demonstrated feasibility of using CLE and topical administration of FITC labeled antibodies for in vivo localization of EGF-R and survivin in esophageal and gastric mucosa.

摘要

目的

本研究旨在建立分子成像的原理,并利用基于探头的共聚焦激光显微内镜(CLE)和局部应用 FITC 标记的抗体,确定猪食管和胃黏膜中表皮生长因子受体(EGF-R)和存活素的体内表达。

方法

本研究在麻醉猪中进行。在胃镜检查期间,将针对 EGF-R 和存活素的 FITC 标记抗体喷涂在食管和胃黏膜的预选区域上,或通过黏膜下注射给药。30 分钟后,使用通过内镜的探头(GastroFlex UHD、Cellvizio、Mauna Kea Technologies、巴黎、法国)进行 pCLE,以确定 EGF-R 和存活素的细胞和组织定位。然后对猪进行安乐死,并收集喷涂或注射抗体的区域的食管和胃壁,用于荧光显微镜下的组织学检查。

结果

CLE 能够可视化食管和胃黏膜中的 EGF-R 和存活素,这通过组织学得到了证实。在食管中,EGF-R 和存活素主要定位于角质形成细胞祖细胞。在胃中,EGF-R 定位于祖细胞区细胞和一些上皮细胞。存活素的定位相似,但涉及更多的表面上皮细胞。本研究证明了使用 CLE 和局部应用 FITC 标记抗体在体内定位食管和胃黏膜中 EGF-R 和存活素的可行性。

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