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蛋白激酶A信号通路参与壳寡糖对猪小肠上皮细胞IPEC-J2的抗炎作用

Involvement of PKA signalling in anti-inflammatory effects of chitosan oligosaccharides in IPEC-J2 porcine epithelial cells.

作者信息

Yang J W, Tian G, Chen D W, Yao Y, He J, Zheng P, Mao X B, Yu J, Huang Z Q, Yu B

机构信息

Institute of Animal Nutrition, Sichuan Agricultural University, Yaan, Sichuan, China.

出版信息

J Anim Physiol Anim Nutr (Berl). 2018 Feb;102(1):252-259. doi: 10.1111/jpn.12686. Epub 2017 Mar 16.

Abstract

Weaning is characterized by intestinal inflammation, which is a big challenge in pig industry. Control of intestinal inflammation is important for improvement of growth performance and health. Therefore, the study was focused on the anti-inflammatory activity of low-molecular-weight chitosan oligosaccharide (LCOS) in a porcine small intestinal epithelial cell line (IPEC-J2). The results showed that TNF-α, as inflammation inducer, significantly upregulated the mRNA expression of IL-8 and MCP-1. Afterwards, LCOS significantly attenuated mRNA expression of IL-8 and MCP-1 induced by TNF-α in the cells. Mannose (MAN), as ligand of mannose receptor, had no effect on the anti-inflammatory activity of LCOS, which suggested that mannose receptor may not involve in the anti-inflammatory activity of LCOS in IPEC-J2 cells. Interestingly, N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide 2HCl hydrate (H89), as PKA (protein kinase A)-specific inhibitor, reversed the mRNA expression of IL-8 when co-cultured with LCOS. Furthermore, LCOS concentration dependent downregulated the mRNA expression of claudin-1 compared with TNF-α treatment. However, the trans-epithelial electric resistance (TEER) was not affected by LCOS when co-cultured with TNF-α in 3 hr. In conclusion, LCOS have a potent anti-inflammatory activity, and as a feed additives, may be useful for the inhibition of inflammatory process in weaning period of pigs with intestinal inflammation occurring.

摘要

断奶的特征是肠道炎症,这是养猪业面临的一大挑战。控制肠道炎症对于提高生长性能和健康状况至关重要。因此,本研究聚焦于低分子量壳寡糖(LCOS)在猪小肠上皮细胞系(IPEC-J2)中的抗炎活性。结果表明,作为炎症诱导剂的TNF-α显著上调了IL-8和MCP-1的mRNA表达。随后,LCOS显著减弱了TNF-α诱导的细胞中IL-8和MCP-1的mRNA表达。甘露糖(MAN)作为甘露糖受体的配体,对LCOS的抗炎活性没有影响,这表明甘露糖受体可能不参与LCOS在IPEC-J2细胞中的抗炎活性。有趣的是,作为蛋白激酶A(PKA)特异性抑制剂的N-[2-(对溴肉桂氨基)乙基]-5-异喹啉磺酰胺二盐酸盐水合物(H89)与LCOS共培养时可逆转IL-8的mRNA表达。此外,与TNF-α处理相比,LCOS浓度依赖性地下调了claudin-1的mRNA表达。然而,与TNF-α共培养3小时时,LCOS对跨上皮电阻(TEER)没有影响。总之,LCOS具有强大的抗炎活性,作为饲料添加剂,可能有助于抑制断奶期发生肠道炎症的仔猪的炎症过程。

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