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

doi:10.16098/j.issn.0529-1356.2021.04.012

解剖学报 ›› 2021, Vol. 52 ›› Issue (4): 574-580.doi: 10.16098/j.issn.0529-1356.2021.04.012

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

杨思宇¹ 杨重锦² 张志欢¹ 穆祥¹ 张永红¹ 张涛^1*

1. 北京农学院动物科学技术学院兽医学（中医药）北京市重点实验室，北京 102206; 2. 北京市神经外科研究所，北京 100070

收稿日期:2020-04-09 修回日期:2020-06-05 出版日期:2021-08-06 发布日期:2021-08-06
通讯作者: 张涛 E-mail:zhangtao@bua.edu.cn
基金资助:
国家自然科学基金;北京农学院大学生科研训练项目;北京市自然科学基金;北京市生猪产业创新团队项目

Determination of carbohydrate chains in two kinds of microvascular endothelial cells by lectin cytochemistry

YANG Si-yu¹ YANG Zhong-jin² ZHANG Zhi-huan¹ MU Xiang¹ ZHANG Yong-hong¹ ZHANG Tao^1*

1.Beijing Key Laboratory of Traditional Chinese Veterinary Medicine, Animal Science and Technology College at Beijing University of Agriculture, Beijing 102206, China;2.Beijing Neurosurgical Institute, Beijing 100070, China

Received:2020-04-09 Revised:2020-06-05 Online:2021-08-06 Published:2021-08-06
Contact: ZHANG Tao E-mail:zhangtao@bua.edu.cn

摘要/Abstract

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Con A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。

关键词: 肺, 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

Abstract:

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 (Con A), phaseolus vulgaris erythroagglutinin (PHA-E), ricinus communis agglutinin Ⅰ (RCA-Ⅰ), lycopersicon esculentum lectin (LEL), sambucus nigra lectin (SNA), ulex europaeus agglutinin Ⅰ (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.

Key words: Lung, Microvascular endothelial cell, Carbohydrate chain, Glycocalyx, Jejunal mucosa, Lectin, Cytochemical staining, Rat, Piglet

中图分类号:

852.1

杨思宇杨重锦张志欢穆祥张永红张涛 . 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[J]. 解剖学报, 2021, 52(4): 574-580.

YANG Si-yu YANG Zhong-jin ZHANG Zhi-huan MU Xiang ZHANG Yong-hong ZHANG Tao. Determination of carbohydrate chains in two kinds of microvascular endothelial cells by lectin cytochemistry[J]. Acta Anatomica Sinica, 2021, 52(4): 574-580.

参考文献

［1］ Kvietys PR, Granger DN. Endothelial cell monolayers as a tool for studying microvascular pathophysiology ［J］. Am J Physiol, 2018, 273(6):G1189-G1199.

［2］ Danese S, Dejana E, Fiocchi C. Immune regulation by microvascular endothelial cells: directing innate and adaptive immunity, coagulation, and inflammation ［J］. J Immunol, 2007, 178(10):6017-6022.

［3］ Reitsma S, Slaaf DW, Vink H, et al. The endothelial glycocalyx: composition, functions, and visualization ［J］. Pflugers Arch, 2007, 454(3):345-359.

［4］ Weinbaum S, Tarbell JM, Damiano ER. The structure and function of the endothelial glycocalyx layer ［J］. Ann Rev Biomed Eng, 2007, 9(1):121-167.

［5］ Hou WF, Yang WG. Sugar chain and proteins glycosylation ［J］. Biotechnology Bulletin, 2005, 21(3):14-17. (in Chinese)

侯温甫，杨文鸽. 糖链及其蛋白质糖基化［J］. 生物技术通报，2005，21(3)：14-17.

［6］ Xiang T, Zhang XL. Glycosylation modification and related functions of virus and host cells ［J］. Progress in Biochemistry and Biophysics, 2017, 44(10):898-907. (in Chinese)

向田，章晓联. 病毒与宿主细胞的糖基化修饰及相关功能［J］. 生物化学与生物物理进展, 2017，44（10）：898-907.

［7］ Xu L, Feng RF, Ma ZhR. Progress on virus receptors ［J］. Progress in Veterinary Medicine, 2015, 36(2):88-92. (in Chinese)

徐雷，冯若飞，马忠仁. 病毒受体研究进展［J］. 动物医学进展，2015, 36(2): 88-92.

［8］ Zhong YG, Qin YN, Sun ShSh, et al. New progress of glycan as receptors for influenza virus ［J］. Progress in Biochemistry and Biophysics. 2012, 39(7): 605-612. (in Chinese)

钟耀刚，秦棪楠，孙士生，等. 流感病毒识别糖链受体分子机制的研究进展［J］. 生物化学与生物物理进展，2012, 39(7): 605-612.

［9］ Langenkamp E, Molema G. Microvascular endothelial cell heterogeneity: general concepts and pharmacological consequences for anti-angiogenic therapy of cancer［J］. Cell Tissue Res, 2009, 335(1):205-222.

［10］ King J, Hamil T, Creighton J, et al. Structural and functional characteristics of lung macro-and microvascular endothelial cell phenotypes ［J］. Microvasc Res, 2004, 67(2):139-151.

［11］ Zhang T, Tian L, Hu G, et al. Microvascular endothelial cells play potential immunoregulatory roles in the immune response to foot-and-mouth disease vaccines［J］. Cell Biochem Funct, 2011, 29(5):394-399.

［12］ Magee JC, Stone AE, Oldham KT, et al. Isolation, culture, and characterization of rat lung microvascular endothelial cells ［J］. Am J Physiol, 1994, 267(4 Pt 1):L433-L441.

［13］ Fehrenbach ML, Cao G, Williams JT, et al. Isolation of murine lung endothelial cells ［J］. Am J Physiol Lung Cell Mol Physiol,2009, 296(6):L1096-L1103.

［14］ Vecchi A, Garlanda C, Lampugnani MG, et al. Monoclonal antibodies specific for endothelial cells of mouse blood vessels. Their application in the identification of adult and embryonic endothelium ［J］. Eur J Cell Biol, 1994, 63(2):247-254.

［15］ Sun X, Yang Z, Mu X, et al. Effects of different fixatives on staining patterns of four lectins in cultured microvascular endothelial cells ［J］. J Histotechnol, 2015, 28(1):22-27.

［16］ Deng YH, Zhou XH, Yang JQ. A lectin histochemical study on the vascular endothelia of rats ［J］. Chinese Journal of Histochemistry and Cytochemistry, 1999, 8(4):394-398. (in Chinese)

邓英华，周新华，杨家齐. SD大鼠血管内皮细胞的凝集素组织化学研究［J］. 中国组织化学与细胞化学杂志, 1999，8（4）：394-398.

［17］ Liew H, Roberts MA, MacGinley R, et al. Endothelial glycocalyx in health and kidney disease: rising star or false Dawn ［J］ ? Nephrology, 2017, 22:940-946.

［18］ Zhang Y, Zhang XY, Liu F, et al. Relationship between terminal sialyl and fucosyl residues of glycans on cell surface and cell biological behaviors ［J］. Acta Biologiae Experimentalis Sinica, 2002, 35(4):271-277. (in Chinese)

张英，张夏英，刘飞，等. 细胞表面糖链末端唾液酸和岩藻糖与某些细胞生物学行为的关系［J］. 实验生物学报，2002，35（4）：271-277.

［19］ Mills AN, Haworth SG. Changes in lectin binding patterns in the developing pulmonary vasculature of the pig lung ［J］. J Pathol, 1986, 149(3):191-199.

［20］ Miyagawa S, Takeishi S, Yamamoto A, et al. Survey of glycoantigens in cells from α1-3 galactosyltransferase knockout pig using a lectin microarray ［J］. Xenotransplantation, 2010, 17(1):61-70.

［21］ Wang Ch, Tian GY. Study on the structure of sugar chains in glycoproteins ［J］. Bulletin of Chemistry, 1994, (5):7-11, 27. (in Chinese)

王晨，田庚元. 糖蛋白中糖链的结构研究［J］. 化学通报，1994，(5)：7-11，27.

［22］ Fischer E, Wagner M, Bertsch T. Cepaea hortensis agglutinin-Ⅰ, specific for oglycosidically linked sialic acids, selectively labels endothelial cells of distinct vascular beds ［J］. Histochem J, 2000, 32(2):105-109.

［23］ Belloni PN, Nicolson GL. Differential expression of cell surface glycoproteins on various organ-derived microvascular endothelial and endothelial cell cultures ［J］. J Cell Physiol, 1988, 136(3):398-410.

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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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