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来自 的 β-葡聚糖参与绵羊瘤胃组织体外免疫刺激。

β-glucan from is involved in immunostimulation of ovine ruminal explants.

机构信息

College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China (Zhang, Jin, Yang, Cao); Animal Medicine College of Henan University of Animal Husbandry and Economy, Zhengzhou, 450046, China (Zhang); Animal Science and Veterinary Medicine College of Henan Agricultural University, Zhengzhou, 450002, China (Jin).

出版信息

Can J Vet Res. 2020 Oct;84(4):283-293.

Abstract

In this study, we investigated whether β-glucan from exerts beneficial effects on mucosal immunity in an ovine ruminal explant (ORE) model. Once the ORE model was established, viability was assessed through histological change, E-cadherin expression, CK-18 and Ki-67 distribution. Then, the OREs were co-cultured with β-glucan, following which, gene and protein expression levels of sheep β-defensin-1 (SBD-1), pro-inflammatory interleukin (IL)-6, and anti-inflammatory IL-10 were detected using quantitative real-time polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA). Hematoxylin & eosin staining, qPCR, and immunohistochemistry showed that the overall ORE structure was intact after 96 hours in culture, but explants cultured for more than 24 hours showed epithelial degradation. Therefore, we performed the follow-up test within 24 hours. qPCR and ELISA revealed that the gene and protein expression levels of SBD-1, IL-6, and IL-10 in the OREs significantly increased ( < 0.05) after treatment with β-glucan compared with controls. This study identified the feasibility and optimal conditions of ORE culture and demonstrated that β-glucan activates SBD-1, IL-6, and IL-10 secretion in OREs to promote mucosal immunity.

摘要

在这项研究中,我们研究了来自 的 β-葡聚糖是否对绵羊瘤胃黏膜免疫产生有益影响。一旦建立了 ORE 模型,就通过组织学变化、E-钙黏蛋白表达、CK-18 和 Ki-67 分布来评估其活力。然后,将 ORE 与 β-葡聚糖共同培养,之后使用实时定量聚合酶链反应(qPCR)和酶联免疫吸附测定(ELISA)检测绵羊 β-防御素-1(SBD-1)、促炎白细胞介素(IL)-6 和抗炎白细胞介素(IL)-10 的基因和蛋白表达水平。苏木精和伊红染色、qPCR 和免疫组织化学显示,培养 96 小时后 ORE 的整体结构完整,但培养超过 24 小时的外植体显示上皮降解。因此,我们在 24 小时内进行了后续测试。qPCR 和 ELISA 显示,与对照组相比,β-葡聚糖处理后 ORE 中 SBD-1、IL-6 和 IL-10 的基因和蛋白表达水平显著增加(<0.05)。本研究确定了 ORE 培养的可行性和最佳条件,并证明 β-葡聚糖激活 ORE 中 SBD-1、IL-6 和 IL-10 的分泌,从而促进黏膜免疫。

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