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两种微血管内皮细胞糖链表达的凝集素细胞化学染色特征

Determination of carbohydrate chains in two kinds of microvascular endothelial cells by lectin cytochemistry
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摘要 目的明确两种微血管内皮细胞(MVECs)糖链表达的特点。方法复苏冻存的大鼠空肠黏膜MVECs;取3日龄SPF仔猪肺组织,采用胶原酶消化法和差速贴壁法分离培养猪肺MVECs;采用凝集素细胞化学方法,检测刀豆凝集素(Con A)、菜豆红细胞凝集素(PHA-E)、蓖麻凝集素Ⅰ(RCA-Ⅰ)、番茄凝集素(LEL)、黑接骨木凝集素(SNA)、荆豆凝集素Ⅰ(UEA-Ⅰ)、麦胚凝集素(WGA)和双花扁豆凝集素(DBA)共8种凝集素在两种MVECs的染色情况。结果大鼠空肠黏膜MVECs对Con A、WGA和LEL呈强阳性着色,PHA-E、SNA和RCA-Ⅰ呈中等阳性着色,DBA呈弱阳性着色,UEA-Ⅰ呈阴性着色;猪肺MVECs对ConA和PHA-E呈强阳性着色,RCA-Ⅰ呈中等阳性着色,LEL和SNA呈弱阳性着色,UEA-Ⅰ、WGA和DBA呈阴性着色。结论两种MVECs的糖链表达具有显著异质性,大鼠空肠黏膜MVECs和猪肺MVECs均不同水平地表达甘露糖、半乳糖、1,3-N-乙酰葡糖胺和唾液酸,N-乙酰葡糖胺和N-乙酰半乳糖胺表达于前者不表达于后者,岩藻糖在两者均不表达。 Objective To determine the expression pattern of carbohydrate chains in two kinds of microvascular endothelial cells(MVECs).Methods Rat jejunal mucosal MVECs were thawed,and lung tissues were removed from specific pathogen free piglet of 3 days old to isolate and culture porcine pulmonary MVECs by collagerase digestion and differential attachment.By lectin cytochemistry,staining of 8 lectins including concanavalin A(ConA),phaseolus vulgaris erythroagglutinin(PHA-E),ricinus communis agglutininⅠ(RCA-Ⅰ),lycopersicon esculentum lectin(LEL),sambucus nigra lectin(SNA),ulex europaeus agglutininⅠ(UEA-Ⅰ),wheat germ agglutinin(WGA)and dolichos biflorus agglutinin(DBA)was detected in rat jejunal mucosal and porcine pulmonary MVECs.Results In rat jejunal mucosal MVECs,strong positve staining was present for Con A,WGA and LEL,medium one for PHA-E、SNA and RCA-Ⅰ,weak one for DBA,and negative staining for UEA-Ⅰ.In porcine pulmonary MVECs,strong positive staining was present for Con A and PHA-E,medium one in RCA-Ⅰ,weak one for LEL and SNA,and negative staining for UEA-Ⅰ,WGA and DBA.Conclusion The carbohydrate patterns in two kinds of MVECs display significant heterogeneity.Both rat jejunal mucosal and porcine pulmonary MVECs express mannose,galactose,1,3-N-acetylglucosamine and sialic acid at different levels.N-acetylglucosamine and N-acetylgalactosamine are detected in the former but not in the latter,and fucose do not in both MVECs.
作者 杨思宇 杨重锦 张志欢 穆祥 张永红 张涛 YANG Si-yu;YANG Zhong-jin;ZHANG Zhi-huan;MU Xiang;ZHANG Yong-hong;ZHANG Tao(Beijing Key Laboratory of Traditional Chinese Veterinary Medicine,Animal Science and Technology College at Beijing University of Agriculture,Beijing 102206,China;Beijing Neurosurgical Institute,Beijing 100070,China)
出处 《解剖学报》 CAS CSCD 北大核心 2021年第4期574-580,共7页 Acta Anatomica Sinica
基金 国家自然科学基金(31672597) 北京农学院大学生科研训练项目(kx2019009) 北京市自然科学基金(6202003) 北京市生猪产业创新团队项目(BAIC02)。
关键词 微血管内皮细胞 糖链 糖萼 空肠黏膜 凝集素 细胞化学染色 大鼠 仔猪 Lung Microvascular endothelial cell Carbohydrate chain Glycocalyx Jejunal mucosa Lectin Cytochemical staining Rat Piglet
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