山豆根多糖对猪圆环病毒Ⅱ型感染免疫细胞增殖活性及炎症相关因子的影响

Effects of Sophora subprostrate polysaccharide on proliferation activity and inflammation-related factors of immune cells infected with porcine circovirusⅡ(PCV2)

【目的】探究山豆根多糖(SSP)对猪圆环病毒Ⅱ型(PCV2)感染RAW264.7细胞增殖活性及炎症相关因子的影响,揭示SSP对PCV2感染免疫细胞炎症相关因子的调控作用。【方法】PCV2体外感染RAW264.7细胞建立炎症模型,以不同浓度(25、50、100、200、400、800和1600μg/mL)SSP进行培养处理,然后采用CCK-8和ELISA分别测定SSP对PCV2体外感染RAW264.7细胞增殖活性及炎症相关因子(IL-1β、IL-8和MCP-1)分泌水平和胞内环氧合酶-1(COX-1)活性的影响。【结果】与细胞对照组相比,SSP浓度≤400μg/mL对RAW264.7细胞增殖活性无显著影响(P>0.05),但SSP浓度达800和1600μg/mL时RAW264.7细胞增殖活性极显著降低(P<0.01,下同),且随培养时间的延长,细胞增殖活性呈先降低后升高的变化趋势,于培养48 h时达最低值。PCV2感染RAW264.7细胞后其增殖活性极显著降低,炎症相关因子IL-1β、IL-8和MCP-1分泌水平及胞内COX-1活性极显著升高;100~400μg/mL SSP能极显著提高PCV2感染RAW264.7细胞增殖活性,且能有效降低RAW264.7细胞的IL-1β、IL-8和MCP-1分泌水平及胞内COX-1活性。具体表现为:与PCV2模型组相比,100μg/mL SSP能显著降低PCV2感染RAW264.7细胞的IL-1β和MCP-1分泌水平(P<0.05,下同);200μg/mL SSP能极显著降低PCV2感染RAW264.7细胞的MCP-1分泌水平,同时显著降低细胞IL-1β和IL-8的分泌水平及胞内COX-1活性;400μg/mL SSP能极显著降低PCV2感染RAW264.7细胞的IL-1β、IL-8和MCP-1分泌水平及胞内COX-1活性。【结论】SSP对RAW264.7细胞增殖活性无显著影响,也未表现出细胞毒性作用,且100~400μg/mL SSP能极显著提高PCV2感染RAW264.7细胞增殖活性,并通过调节PCV2感染免疫细胞的炎症相关因子水平而发挥抗炎作用。

【Objective】This study aimed to investigate the proliferation activity and inflammation-related factors of Sophora subprostrate polysaccharide(SSP)on porcine circovirusⅡ(PCV2)infected RAW264.7 cells,and to reveal its regulation on PCV2 infected immune cells inflammation-related factors.【Method】Established the inflammatory model of PCV2 infected RAW264.7 cells in vitro,and then cultured with different concentrations(25,50,100,200,400,800 and 1600μg/mL)of SSP.CCK-8 method was used to determine the proliferation activity of RAW264.7 cells and PCV2 infected RAW264.7 cells with SSP at different concentrations.The effect of SSP at different concentrations on inflammationrelated factors(IL-1β,IL-8,MCP-1 and COX-1)in PCV2 infected RAW264.7 cells were determined by enzyme linked immunosorbent assay(ELISA).【Result】Compared with the cell control group,the results showed that the concentration of SSP≤400μg/mL had no significant effect on RAW264.7 cell viability(P>0.05),but SSP concentrations at 800μg/mL and 1600μg/mL extremely significantly decreased the viability of RAW264.7 cells(P<0.01,the same below).After PCV2 infection,the proliferation activity of RAW264.7 cells was extremely decreased,the secretion levels of inflammatory cytokines IL-1β,IL-8 and MCP-1 and the activity of intracellular COX-1 were extremely increased.100-400μg/mL of SSP exttemely increased the viability of PCV2 infected RAW264.7 cells,and the secretion levels of IL-1β,IL-8,MCP-1 and the activity of COX-1 in RAW264.7 cells were decreased compared to PCV2 group.Compared with PCV2 model group,100μg/mL of SSP significantly decreased IL-1βand MCP-1 secretion levels in PCV2 infected RAW264.7 cells(P<0.05,the same below);200μg/mL of SSP extremely decreased the secretion level of MCP-1,significantly decreased the secretion levels of IL-1βand IL-8,and the activity of COX-1 in PCV2 infected RAW264.7 cells;SSP at 400μg/mL of could extremely reduce the levels of IL-1β,IL-8 and MCP-1 and the activity of COX-1 in PCV2 infected RAW264.7 cells.【Conclusion】SSP has no significant effect on the proliferation activity of RAW264.7 cells,and shows no cytotoxicity.100-400μg/mL of SSP extremely increases the proliferation activity of PCV2 infected RAW264.7 cells,and play an anti-inflammatory effect via regulating the secretion levels of inflammation-related factors.