猴头菇多糖对MDRV感染RAW264.7细胞TLR3/TRIF诱导表达及病毒复制的影响

doi:10.16431/j.cnki.1671-7236.2021.09.033

Abstract

Abstract: The aim of this study was to investigate the effect of Hericium erinaceus polysaccharide (HEP) pretreatment of RAW264.7 cells on the replication of Muscovy duck reovirus (MDRV) and its mechanism, and dissect the mechanism of HEP in regulating MDRV-induced immunosuppression in Muscovy ducks from the Toll-like receptor 3/interferon-β TIR domain adaptor protein (TLR3/TRIF) signaling pathway. The trial included blank control group (BCG), viral infection control group (VCG), HEP control group (HCG) and HEP prevention viral infection group (HPG). The protein expression levels of TLR3, TRIF and tumor necrosis factor receptor-associated factor 6 (TRAF6) in RAW264.7 cell at 12 and 24 h of MDRV infection was detected by Western blotting. After virus infection for 24 h, the contents of tumor necrosis factor α (TNF-α), interleukin 10 (IL-10), IL-6, IL-1β and interferon β (IFN-β) in the cell supernatant culture medium of each group were detected by ELISA. The TCID₅₀ detection results showed that the TCID₅₀ of MDRV was 10^3.46. At 12 h after infection, the cells did not change significantly, 24 h after infection, the cells became round, and 36 h after infection, a large number of cells died. The replication results of MDRV in RAW264.7 cells showed that within 12-24 h of virus infection, the expression of σNS increased, and at 24-36 h, the expression of σNS was maintained at a high level. Western blotting results showed that compared with blank control group, the expression levels of TLR3, TRIF and TRAF6 proteins in VCG group were significantly increased (P<0.05) after 12 and 24 h infection, and TRIF protein in HCG group were significantly increased (P<0.05) after 24 h infection. When compared with infected group, the expression of σNS, TLR3, TRIF and TRAF6 proteins were significantly decreased (P<0.05) after 12 and 24 h infection in the HPG group. ELISA results showed that compared with blank control group, the contents of TNF-α, IL-6, IL-1β and IFN-β in the culture medium of VCG group were significantly increased (P<0.05). When compared with infected group, the contents of TNF-α, IL-6 and IL-1β in the cell culture medium of HPG group were significantly decreased (P<0.05), while the content of IFN-β was significantly increased (P<0.05). The results showed that HEP could regulate MDRV infection to induce TLR3 signal transduction pathway activation, inhibit the overexpression of TNF-α, IL-10, IL-6 and IL-1β of downstream products of TLR3 signal transduction pathway, and up-regulate the expression of IFN-β in RAW264.7 cells, thereby inhibiting the replication of MDRV in RAW264.7 cells.

Key words: Muscovy duck reovirus; Hericium erinaceus polysaccharide; Toll-like receptor 3; signal transduction; virus replication

CLC Number:

S858.32

YAN Ping, LIN Shaoqing, LI Minghui, SUN Xuening, LI Jian, HUANG Yifan, WU Yijian. Effects of Hericium erinaceus Polysaccharide on TLR3/TRIF-induced Expression and Viral Replication in MDRV-infected RAW264.7 Cells[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(9): 3415-3422.

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Effects of Hericium erinaceus Polysaccharide on TLR3/TRIF-induced Expression and Viral Replication in MDRV-infected RAW264.7 Cells

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