旋毛虫重组二肽基肽酶1在体外对大鼠外周血单个核细胞增殖、吞噬、凋亡等的影响

doi:10.11843/j.issn.0366-6964.2020.019

摘要/Abstract

摘要： 本文旨在分析旋毛虫二肽基肽酶1（dipeptidyl-peptidase 1，Dpp1）在体外对大鼠外周血单个核细胞（peripheral blood mononuclear cells，PBMCs）增殖、迁移、凋亡以及一氧化氮（NO）分泌和吞噬功能的影响。根据GenBank中旋毛虫Dpp1基因序列设计一对特异性引物，RT-PCR获得该基因，将其亚克隆到pET-32a原核表达载体，IPTG诱导后获得重组蛋白rDpp1。将大鼠PBMCs与rDpp1共孵育，用间接免疫荧光试验（IFA）分析rDpp1与大鼠PBMCs的结合情况，分析不同质量浓度（10、20、40 μg·mL^-1）的rDpp1对细胞增殖、迁移、NO分泌、吞噬功能和细胞凋亡的影响。结果表明，40 μg·mL^-1rDpp1极显著促进PBMCs增殖；10 μg·mL^-1rDpp1能显著促进PBMCs迁移，而20及40 μg·mL^-1时表现极显著的促进；20与40 μg·mL^-1rDpp1极显著促进PBMCs分泌NO；10 μg·mL^-1rDpp1能显著促进PBMCs的吞噬功能，而20及40 μg·mL^-1时表现极显著的促进；各质量浓度蛋白均极显著地促进PBMCs的凋亡。旋毛虫二肽基肽酶1在体外能通过多种途径影响大鼠外周血单个核细胞，对调节宿主免疫应答有一定的促进作用。

关键词: 旋毛虫, 二肽基肽酶1, 外周血单个核细胞, 吞噬, 凋亡

Abstract: This research aimed to investigate the effect of Trichinella spiralis dipeptidyl-peptidase 1 (Dpp1) on rat peripheral blood mononuclear cell (PBMC) proliferation, migration, apoptosis, nitric oxide secretion and phagocytosis in vitro. A pair of primers were designed based on Dpp1 gene nucleotides in GenBank, Dpp1 gene was cloned by RT-PCR and the expression vector was constructed to obtain the recombinant protein (rDpp1). The reactogenicity of rDpp1 was analyzed by Western blot. PBMCs were separated from rats and co-incubated with rDpp1 in vitro,binding of rDpp1 with rat PBMCs was confirmed by immunofluorescence assay (IFA), and the effects of different concentration of rDpp1 (10, 20, 40 μg·mL^-1) on cell proliferation,migration,nitric oxide production,phagocytosis and apoptosis were determined. The results showed that rDpp1 at a concentration of 40 μg·mL^-1 significantly promoted the proliferation of PBMCs; rDpp1 at 10 μg·mL^-1 could promote cell migration, while 20 and 40 μg·mL^-1 concentrations showed a significant promotion; rDpp1 with 40 μg·mL^-1 concentration significantly promoted NO secretion; rDpp1 with 10 μg·mL^-1 promoted cell phagocytosis, and 20 and 40 μg·mL^-1 concentrations showed very significant promotion; each concentration of proteins could significantly promote the apoptosis of PBMCs. The functions of rat PBMCs were influenced by T. spiralis dipeptidyl-peptidase 1 via different pathways.

Key words: Trichinella spiralis, dipeptidyl-peptidase 1, peripheral blood mononuclear cells, phagocytosis, apoptosis

中图分类号:

廖书漪, 储稳, 徐立新, 宋小凯, 李祥瑞, 严若峰. 旋毛虫重组二肽基肽酶1在体外对大鼠外周血单个核细胞增殖、吞噬、凋亡等的影响[J]. 畜牧兽医学报, 2020, 51(6): 1362-1370.

LIAO Shuyi, CHU Wen, XU Lixin, SONG Xiaokai, LI Xiangrui, YAN Ruofeng. Effect of Trichinella spiralis Recombinant Dipeptidyl-peptidase 1 on Proliferation, Phagocytosis, and Apoptosis of Rat PBMCs in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(6): 1362-1370.

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