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多种凝集素对大鼠胃黏膜不同层来源黏蛋白的反应:酶联凝集素结合测定法与凝集素组织化学的比较

Reaction of various lectins to mucin derived from the different layers of rat gastric mucosa: comparison of enzyme-linked lectin binding assay with lectin histochemistry.

作者信息

Kodaira H, Ishihara K, Hotta K, Kagoshima M, Shimada H, Ishii K

机构信息

Department of Molecular Pharmacology, School of Pharmaceutical Sciences, Kitasato University, Tokyo, Japan.

出版信息

Biol Pharm Bull. 2000 Oct;23(10):1173-9. doi: 10.1248/bpb.23.1173.

Abstract

The purpose of this study was to demonstrate that the histochemical staining reactivities of lectins in rat stomach actually represent the gastric mucins, and to estimate the utility of the lectins for mucin histochemistry. In this paper, the lectin histochemistry was compared with an enzyme-linked lectin binding assay (ELLA) of the mucins derived from distinct regions and layers of the Sprague-Dawley rat stomach and it was examined to determine the definite binding and problematic binding of the conventional lectin. Among the 10 different biotinylated lectins, Canavalia ensiformis (ConA), Griffonia simplicifolia II (GS-II), Triticuin vulgaris (WGA), Ricinus communis I (RCA-I), Arachis hypogaea (PNA), Glycine max (SBA), Dolichos biflorus (DBA), Ulex europaeus I (UEA-I), Sambucus nigra (SNA) and Maackia amurensis II (MAL-II), examined in this study, GS-II, SBA, DBA, UEA-I, SNA and MAL-II bound clearly to the mucin of distinct regions and layers of the Sprague-Dawley rat stomach in agreement with the results of ELLA. Namely GS-II lectins preferentially bound to the mucin in the mucous neck cells of the corpus area. SBA and DBA clearly recognized the mucin in the covering epithelial mucous cells in the corpus and antral area. UEA-I was widely bound to all the mucin present in both the corpus and antrum. On the other hand, SNA and MAL-II could not react with the mucin obtained from the gastric mucosa but was specifically bound to the mucin purified from the mucous gel layer. These results suggested that the lectins described above are useful histochemical tools to recognize the mucus present in the different regions and layers of Sprague-Dawley rat gastric mucosa.

摘要

本研究的目的是证明大鼠胃中凝集素的组织化学染色反应性实际上代表胃粘蛋白,并评估凝集素在粘蛋白组织化学中的效用。本文将凝集素组织化学与源自Sprague-Dawley大鼠胃不同区域和层的粘蛋白的酶联凝集素结合测定(ELLA)进行了比较,并检测了传统凝集素的确定结合和有问题的结合。在本研究中检测的10种不同的生物素化凝集素中,即刀豆球蛋白A(ConA)、西非单叶豆凝集素II(GS-II)、小麦胚芽凝集素(WGA)、蓖麻凝集素I(RCA-I)、花生凝集素(PNA)、大豆凝集素(SBA)、双花扁豆凝集素(DBA)、荆豆凝集素I(UEA-I)、黑接骨木凝集素(SNA)和山槐凝集素II(MAL-II),GS-II、SBA、DBA、UEA-I、SNA和MAL-II与Sprague-Dawley大鼠胃不同区域和层的粘蛋白有明显结合,与ELLA结果一致。即GS-II凝集素优先结合于胃体部粘液颈细胞中的粘蛋白。SBA和DBA能清楚地识别胃体和胃窦部覆盖上皮粘液细胞中的粘蛋白。UEA-I广泛结合于胃体和胃窦中存在的所有粘蛋白。另一方面,SNA和MAL-II不能与从胃粘膜获得的粘蛋白反应,但能特异性结合从粘液凝胶层纯化的粘蛋白。这些结果表明,上述凝集素是识别Sprague-Dawley大鼠胃粘膜不同区域和层中存在的粘液的有用组织化学工具。

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