辣蓼黄酮乙酸乙酯部分对PRV体外诱导RAW264.7细胞分泌炎性因子的影响

doi:10.16431/j.cnki.1671-7236.2021.03.038

摘要/Abstract

摘要： 探讨辣蓼黄酮乙酸乙酯部分（ethyl acetate of flavonoids from Polygonum hydropiper L.，FEA）对猪伪狂犬病病毒（Pseudorabies virus，PRV）体外诱导RAW264.7细胞分泌炎性因子的影响。正式试验前将不同浓度的FEA作用于RAW264.7细胞，通过CCK-8检测细胞体外增殖活性，筛选FEA对RAW264.7细胞的安全浓度范围。正式试验分为空白对照组、PRV阳性对照组、芦丁对照组、5个FEA药物组，除空白对照组外，其余各组细胞加入PRV共孵育1.5 h后，再加入芦丁或不同浓度的FEA，分别培养4、8、12、24 h，CCK-8法检测FEA对病毒感染的RAW264.7细胞活性，筛选出3个FEA药物组（高、中、低剂量FEA），再通过ELISA法检测各时间点细胞培养液上清中炎症相关因子TNF-α、IL-1β、IL-6、IL-10、MCP-1和IFN-γ的分泌水平，以确定FEA体外调节炎症反应的最适时间及药物浓度。结果显示：①FEA对RAW264.7细胞的安全浓度为12.5~200 μg/mL；②25~100 μg/mL FEA能显著提高PRV感染的RAW264.7细胞体外增殖活性；③PRV感染RAW264.7细胞后促进细胞炎症因子的分泌，用FEA处理8~12 h后，在一定程度上降低了TNF-α、IL-1β、IL-6和MCP-1的分泌水平（P<0.05），升高了IFN-γ分泌水平（P<0.05），调节了IL-10分泌水平，且FEA处理8 h抗炎效果最好。结果表明，FEA能调节病毒感染细胞分泌炎性因子的水平，提示其具有体外抗炎作用。该结果可为进一步研究FEA抗病毒分子机制提供参考依据。

关键词: 辣蓼黄酮乙酸乙酯部分(FEA); 伪狂犬病病毒; 炎性因子

Abstract: The present study investigated the effect of FEA on inflammatory cytokines secretion in PRV induced RAW264.7 cells.Briefly,CCK-8 method was used to detect the activity of cells in vitro,in order to screen the safe concentrations of FEA on RAW264.7 cells.Then,the groups were assigned as blank control group,PRV positive group,rutin group and five FEA groups.The RAW264.7 cells(except for blank control group)were incubated with PRV for 1.5 h,then cultured with different concentrations of rutin or FEA for 4,8,12 and 24 h,respectively.CCK-8 method was used to detect the activity of PRV induced cells for screen three concentrations of FEA (high,medium and low doses)and then ELISA method were used to detect the secretion levels of TNF-α,IL-1β,IL-6,IL-10,MCP-1 and IFN-γ,in order to evaluate the optimal time and concentration of FEA for anti-inflammatory effect.The results showed as follows:①The safe concentration of FEA was ranged from 12.5 to 200 μg/mL.②In vitro,25-100 μg/mL of FEA could significantly improve the proliferation activity of PRV-induced RAW264.7 cell.③The levels of inflammatory cytokines were increased in PRV induced RAW264.7 cells,treatment of FEA for 8-12 h markedly decreased the levels of TNF-α,IL-1β,IL-6 and MCP-1 (P<0.05),increased the level of IFN-γ when compared with PRV group (P<0.05),and the IL-10 level was also regulated by FEA to a certain extent.Moreover,the anti-inflammatory effect was much better post FEA treatment for 8 h.In conlusion,FEA could regulate the secretion levels of inflammatory cytokines in PRV-induced RAW264.7 cell and exert anti-inflammatory effects in vitro.The results provided reference for further research on the molecular mechanism for antiviral drug.

Key words: ethyl acetate of flavonoids from Polygonum hydropiper L.(FEA); Pseudorabies virus; inflammatory cytokines

中图分类号:

S859.7

任春芝, 路彩霞, 韦英益, 于美玲, 胡庭俊. 辣蓼黄酮乙酸乙酯部分对PRV体外诱导RAW264.7细胞分泌炎性因子的影响[J]. 中国畜牧兽医, 2021, 48(3): 1132-1140.

REN Chunzhi, LU Caixia, WEI Yingyi, YU Meiling, HU Tingjun. Effect of Ethyl Acetate Fraction of Flavonoids from Polygonum hydropiper L. on Cytokines Secretion in RAW264.7 Cells Induced by Pseudorabies Virus[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(3): 1132-1140.

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