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甘草酸抑制 LPS 诱导的体外山羊瘤胃上皮细胞炎症反应。

Glycyrrhizin inhibits LPS-induced inflammatory responses in goat ruminal epithelial cells in vitro.

机构信息

College of Animal Science and Technology, Tarim University, Alar, 843300, Xinjiang, PR China.

Department of Clinical Veterinary Medicine, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, Hubei, PR China.

出版信息

BMC Mol Cell Biol. 2023 Sep 19;24(1):28. doi: 10.1186/s12860-023-00489-y.

Abstract

Inflammation plays a crucial role in the progression of Subacute Ruminal Acidosis (SARA). The experiment was designed to investigate anti-inflammatory effects of glycyrrhizin on goats ruminal epithelial cells (GREC) which were induced SARA by Lipopolysaccharide (LPS) in vitro. The GREC were induced SARA by adding LPS at the concentration of 5 μm and glycyrrhizin was added at different concentration of 0, 60, 90, 120, 150 μm. The structural integrity of LPS-induced GREC with the treatment of glycyrrhizin were observed by electron microscope; The levels of inflammatory factors TNF-α, IL-1β, IL-6, IL-8 and IL-12 were measured by ELISA; The number of Zo-1 and Occludin were measured, the expression of tight junction protein Occludin were measured by Western blot, and the mRNA expression of NF-κB, TNF-α, IL-1β, IL-6, IL-8 and IL-12 were measured in vitro. The results showed that higher concentration treatment of glycyrrhizin led to better morphology in LPS-induced GREC. Glycyrrhizin inhibited the growth of inflammatory factors TNF-α, IL-1β, IL-6, IL-8 and IL-12 in a dose-dependent manner. The number of ZO-1 and Occludin increased with the increase of adding of glycyrrhizin. Western blot analysis showed that the expression of tight junction protein Occludin in LPS-induced GREC increased with the adding of glycyrrhizin in a dose-dependent manner. Furthermore, the mRNA expression of NF-κB, TNF-α, IL-1β, IL-6, IL-8 and IL-12 decreased significantly with the increase treatment of glycyrrhizin. Glycyrrhizin significantly inhibits LPS-induced inflammatory mediators in GREC and the effects are better with the increase treatment of glycyrrhizin in vitro.

摘要

炎症在亚急性瘤胃酸中毒(SARA)的发展中起着至关重要的作用。本实验旨在研究甘草酸对脂多糖(LPS)体外诱导的山羊瘤胃上皮细胞(GREC)的抗炎作用。通过添加浓度为 5 μm 的 LPS 诱导 GREC 发生 SARA,同时添加不同浓度的甘草酸(0、60、90、120、150 μm)。通过电子显微镜观察经甘草酸处理的 LPS 诱导的 GREC 的结构完整性;通过 ELISA 测量炎性因子 TNF-α、IL-1β、IL-6、IL-8 和 IL-12 的水平;测量 Zo-1 和 Occludin 的数量,通过 Western blot 测量紧密连接蛋白 Occludin 的表达,以及测量 NF-κB、TNF-α、IL-1β、IL-6、IL-8 和 IL-12 的体外 mRNA 表达。结果表明,甘草酸的高浓度处理导致 LPS 诱导的 GREC 形态更好。甘草酸以剂量依赖的方式抑制炎性因子 TNF-α、IL-1β、IL-6、IL-8 和 IL-12 的生长。随着甘草酸添加量的增加,ZO-1 和 Occludin 的数量增加。Western blot 分析表明,随着甘草酸添加量的增加,LPS 诱导的 GREC 中紧密连接蛋白 Occludin 的表达呈剂量依赖性增加。此外,随着甘草酸处理量的增加,NF-κB、TNF-α、IL-1β、IL-6、IL-8 和 IL-12 的 mRNA 表达显著下降。甘草酸可显著抑制 LPS 诱导的 GREC 中炎性介质的表达,且体外作用随甘草酸处理量的增加而增强。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bb2/10507872/bdf5f8a6b434/12860_2023_489_Fig2_HTML.jpg

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