为探究板蓝根多糖(Radix isatidis polysaccharide,IRPS)对猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)感染后细胞因子分泌的影响.本试验用永生化的猪肺泡巨噬细胞(3D4/21/CD163)作为靶细胞,分别采取ELISA和 Western blot等试验方法对其炎性细胞因子以及诱导炎性细胞因子分泌的TLR4和NF-κB等蛋白表达水平进行测定,并通过TAK-242(TLR4特异性抑制剂)对TLR4通路进行抑制/封闭,ELISA检测TLR4-NF-κB信号通路被阻断后的IRPS对PRRSV感染3D4/21/CD163细胞分泌细胞因子的影响.结果显示IRPS能够降低PRRSV感染3D4/21/CD163细胞后IL-1β、1L-6、IL-8和TNF-α等炎性细胞因子的分泌,并且能够降低PRRSV感染3D4/21/CD163细胞后TLR4和NF-κB的蛋白表达水平;当TLR4-NF-KB信号通路被阻断时,炎性细胞因子的表达相对于单独的IRPS组有不同程度的升高.说明IRPS能够通过TLR4-NF-κB信号通路影响炎性细胞因子的分泌,减少炎性细胞因子对细胞的损伤.
Effects of IRPS on cytokine secretion after PRRSV infection via TLR4-NF-κB sig-naling pathway
Immortalized porcine alveolar macrophages(3D4/21/CD163)were used as target cells to study the effect of Radix isatidis polysaccharide(IRPS)on cytokine secretion after porcine repro-ductive and respiratory syndrome virus(PRRSV)infection.ELISA and Western blot were used to detect the levels of inflammatory cytokines,TLR4,and NF-κB protein.After TLR4 blocking with TAK-242(TLR4 specific inhibitor),the effect of IRPS on cytokine secretion after PRRSV infec-tion of 3D4/21/CD163 cells was measured by ELISA.The results showed that IRPS reduced the secretion of inflammatory cytokines such as IL-1β,IL-6,IL-8,and TNF-α,and reduced the protein expression of TLR4 and NF-κB in 3D4/21/CD163 cells after PRRSV infection.When the TLR4-NF-κB signaling pathway was blocked by TAK-242,the secretion of inflammatory factors in-creased compared with the IRPS group unbroken.It shows that IRPS can affect the secretion of in-flammatory cytokines through the TLR4-NF-κB signaling pathway and reduce the damage of in-flammatory cytokines to cells.
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