The Effect of Clostridium butyricum-Derived Lipoteichoic Acid on Lipopolysaccharide-Stimulated Porcine Intestinal Epithelial Cells.

IF 1.8 3区农林科学 Q2 VETERINARY SCIENCES

Veterinary Medicine and Science Pub Date : 2025-01-01 DOI:10.1002/vms3.70157

Qu Chen, Lingyan Ma, Yang Wen, Wentao Lyu, Minjie Yu, Hua Yang, Yingping Xiao

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引用次数: 0

Abstract

Background: Clostridium butyricum is a probiotic widely used in animal husbandry, and there is evidence to suggest that it can alleviate intestinal inflammation in pigs and may be related to its lipoteichoic acid (LTA), but the mechanism is still unclear.

Objective: This study aimed to determine the regulatory effect and potential mechanism of C. butyricum LTA on LPS-stimulated inflammation in intestinal porcine epithelial line-J2 (IPEC-J2).

Methods: IPEC-J2 cells were treated with LPS and different concentrations of LTA (0.05, 0.1 and 0.15 mM). After treatment of 0.5, 1.5 and 4.5 h, the cell culture media were collected for the measurement of TNF-α and IL-10 by using ELISA kits, and the cells were collected for RT-qPCR and Western blotting detections. Further elucidating the pathway of LTA regulating IL-10 and TNF-α gene expression by inhibiting key proteins in the toll-like receptor pathway with antagonists C34, PDTC, SB230580 and U0126.

Results: High-dose LTA significantly promoted the secretion of the anti-inflammatory factor IL-10 in IPEC-J2 cells, and inhibited the expression and secretion of pro-inflammatory TNF-α in the short term. LTA inhibited the gene expression of TLR4 in LPS-stimulated cells and reduced the protein phosphorylation levels of p38, ERK1/2 and p65. The inhibition of TLR4, p38, ERK1/2 and p65 reduced the TNF-α gene expression caused by LPS; LTA increased TLR2 gene expression, inhibition of p38, ERK and p65 rather than TLR4 reduced the IL-10 gene expression.

Conclusion: Our study found that C. butyricum LTA was an important component of C. butyricum regulating the inflammatory response of IPEC-J2 cells. LTA mainly reduced the expression of TNF-α by inhibiting TLR4, while stimulating TLR2 increased the expression of IL-10. Downstream p65, p38 and ERK1/2 were involved in regulating both TNF-α and IL-10. However, TLR4 was only related to the increase in TNF-α caused by LPS and not to the increase in IL-10 caused by LTA. Our work supplemented the probiotic mechanism of C. butyricum and provided a theoretical basis for the application of C. butyricum LTA.

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丁酸梭菌衍生脂壁酸对脂多糖刺激的猪肠上皮细胞的影响。

背景：丁酸梭菌（Clostridium butyricum）是一种广泛应用于畜牧业的益生菌，有证据表明它可以缓解猪的肠道炎症，可能与其脂磷胆酸（LTA）有关，但其机制尚不清楚。目的：研究丁酸梭菌LTA对lps刺激的猪肠上皮细胞系j2 （IPEC-J2）炎症的调节作用及其可能机制。方法：LPS和不同浓度LTA（0.05、0.1、0.15 mM）处理IPEC-J2细胞。处理0.5、1.5、4.5 h后，收集细胞培养液，采用ELISA试剂盒检测TNF-α和IL-10，收集细胞进行RT-qPCR和Western blotting检测。通过拮抗剂C34、PDTC、SB230580和U0126，进一步阐明LTA通过抑制toll样受体通路关键蛋白调控IL-10和TNF-α基因表达的途径。结果：大剂量LTA显著促进IPEC-J2细胞抗炎因子IL-10的分泌，短期内抑制促炎因子TNF-α的表达和分泌。LTA抑制lps刺激细胞中TLR4基因表达，降低p38、ERK1/2和p65蛋白磷酸化水平。抑制TLR4、p38、ERK1/2和p65可降低LPS引起的TNF-α基因表达；LTA增加TLR2基因表达，抑制p38、ERK和p65而非TLR4降低IL-10基因表达。结论：本研究发现丁酸梭菌LTA是丁酸梭菌调节IPEC-J2细胞炎症反应的重要成分。LTA主要通过抑制TLR4降低TNF-α的表达，而刺激TLR2增加IL-10的表达。下游的p65、p38和ERK1/2参与调节TNF-α和IL-10。而TLR4仅与LPS引起的TNF-α升高有关，与LTA引起的IL-10升高无关。本研究补充了丁酸梭菌的益生菌机制，为丁酸梭菌LTA的应用提供了理论依据。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Veterinary Medicine and Science Veterinary-General Veterinary

CiteScore

3.00

自引率

0.00%

发文量

296

期刊介绍： Veterinary Medicine and Science is the peer-reviewed journal for rapid dissemination of research in all areas of veterinary medicine and science. The journal aims to serve the research community by providing a vehicle for authors wishing to publish interesting and high quality work in both fundamental and clinical veterinary medicine and science. Veterinary Medicine and Science publishes original research articles, systematic reviews, meta-analyses, and research methods papers, along with invited editorials and commentaries. Original research papers must report well-conducted research with conclusions supported by the data presented in the paper. We aim to be a truly global forum for high-quality research in veterinary medicine and science, and believe that the best research should be published and made widely accessible as quickly as possible. Veterinary Medicine and Science publishes papers submitted directly to the journal and those referred from a select group of prestigious journals published by Wiley-Blackwell. Veterinary Medicine and Science is a Wiley Open Access journal, one of a new series of peer-reviewed titles publishing quality research with speed and efficiency. For further information visit the Wiley Open Access website.