黄芪多糖诱导SOCS3表达对鸡巨噬细胞炎症反应的抑制作用

doi:10.16431/j.cnki.1671-7236.2021.11.038

Abstract

Abstract: The aim of this study was to investigate the anti-inflammatory effect and action mechanism of Astragalus polysaccharide (APS) on LPS-induced chicken macrophage (HD11) inflammation model. HD11 cells were stimulated with different concentrations of LPS. Cell viability was detected by CCK8 assay and interleukin-1β(IL-1β) mRNA expression was detected by Real-time quantitative PCR in order to determine the optimal concentration of LPS for the construction of cellular inflammation model. HD11 cells were divided into control group (C), lipopolysaccharide (LPS) group (L), APS group (A) and APS inhibited LPS group (A+L). The mRNA expressions of IL-1β, tumor necrosis factor-α (TNF-α), NF-κBp65, p38MAPK and suppresser of cytokine signaling 3 (SOCS3) were detected by Real-time quantitative PCR, the protein contents of NF-κBp65, p38MAPK and SOCS3 were detected by Western blotting, and the protein contents of IL-1β and TNF-α were detected by ELISA at 2, 6, 12, 24 and 48 h after LPS stimulation. The results of cell viability and IL-1β detection showed that the optimal addition concentration of LPS for construction of cellular inflammation model was 0.5 μ g/mL LPS. Real-time quantitative PCR results showed that compared with control group, the mRNA expressions of IL-1β, TNF-α, NF-κBp65, p38MAPK and SOCS3 in L and A groups were significantly increased (P<0.05). Compared with L group, the mRNA expression levels of IL-1β, TNF-α, NF-κBp65 and p38MAPK in A+L group were significantly decreased (P<0.05), and the mRNA expression level of SOCS3 was significantly increased (P<0.05). Western blotting results showed that compared with control group, the ratios of P-NF-κBp65/NF-κBp65, P-p38MAPK/p38MAPK and SOCS3/α-tubulin in A group were significantly increased (P<0.05). Compared with L group, the ratios of P-NF-κBp65/NF-κBp65 and P-p38MAPK/p38MAPK in A+L group were significantly decreased (P<0.05), and SOCS3/α-tubulin ratio in A+L group was significantly increased (P<0.05). ELISA results showed that compared with control group, the protein contents of IL-1β and TNF-α in L group were significantly increased within 2-48 h (P<0.05), the protein contents of IL-1β and TNF-α in A+L group were significantly decreased within 12-48 h after LPS stimulation when compared with L group (P<0.05). The results suggested that in the LPS-induced HD11 cell inflammation model, APS could inhibit the overactivation of NF-κBp65 and p38MAPK signaling pathways by promoting SOCS3 expression, and thus played an anti-inflammatory role.

Key words: Astragalus polysaccharide; anti-inflammatory; suppresser of cytokine signaling 3 (SOCS3)

CLC Number:

S831.7

TAO Xinlei, LIU Danhua, TIAN Xu, ZHENG Shimin, GAO Xueli, LYU Xiaoping, LIU Chaonan. Inhibitory Effect of Astragalus Polysaccharide on Inflammatory Response of Chicken Macrophages by Inducing SOCS3 Expression[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 4284-4291.

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Inhibitory Effect of Astragalus Polysaccharide on Inflammatory Response of Chicken Macrophages by Inducing SOCS3 Expression

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