山豆根多糖对猪圆环病毒Ⅱ型感染猪肺泡巨噬细胞炎性因子水平的影响

doi:10.16431/j.cnki.1671-7236.2021.05.034

摘要/Abstract

摘要： 试验旨在研究山豆根多糖（SSP）对猪肺泡巨噬细胞3D4/2感染猪圆环病毒Ⅱ型（Porcine circovirus virus Ⅱ type，PCV2）后炎性因子水平的影响。本试验设空白对照组、PCV2感染组、脂多糖（LPS）对照组和3种浓度（100、200和400 μg/mL）的山豆根多糖组，采用PCV2体外感染猪肺泡巨噬细胞，在培养液中分别加入100、200和400 μg/mL山豆根多糖培养24 h后收集样品，用酶联免疫吸附法（ELISA）测定细胞分泌单核细胞趋化蛋白-1（MCP-1）、白细胞介素-8（IL-8）水平和胞内环氧合酶-1（COX-1）、环氧合酶-2（COX-2）的酶活性，实时荧光定量PCR和Western blotting法分别检测诱导型一氧化氮合酶（iNOS）、COX-2基因及其蛋白表达。结果显示，PCV2感染猪肺泡巨噬细胞后极显著提高了MCP-1、IL-8分泌水平和细胞内COX-1、COX-2酶活性（P<0.01），iNOS、COX-2 mRNA和蛋白表达水平均极显著提升（P<0.01）；100、200、400 μg/mL山豆根多糖处理后，3D4/2细胞MCP-1、IL-8分泌水平和细胞内COX-1、COX-2酶活性均显著或极显著降低（P<0.05；P<0.01），其中400 μg/mL山豆根多糖处理后效果最显著，细胞内iNOS、COX-2基因mRNA表达水平同样显著或极显著降低（P<0.05；P<0.01）；100、200、400 μg/mL山豆根多糖处理后，显著或极显著抑制了由PCV2诱导的iNOS蛋白表达量的增加（P<0.05；P<0.01），经200、400 μg/mL山豆根多糖处理后显著抑制COX-2蛋白表达量的增加，400 μg/mL山豆根多糖处理效果最佳。综上所述，山豆根多糖可在一定程度上抑制PCV2感染所引起的促炎症因子mRNA表达的升高及其蛋白的表达，起到一定的缓解炎症的作用。

关键词: 山豆根多糖; 猪圆环病毒Ⅱ型; 猪肺泡巨噬细胞3D4/2; 炎性因子

Abstract: To investigate the effect of Sophora subprosrate polysaccharide (SSP) on the levels of inflammatory factors in 3D4/2 cells infected by Porcine circovirus virus Ⅱ type (PCV2),in this study,blank control group,PCV2 group,lipopolysaccharide (LPS) group and three concentrations of SSP group (100,200 and 400 μg/mL) were established,respectively.3D4/2 cells was infected by PCV2 in vitro, and then the cells were treated with different concentrations of SSP for 24 h.The secretions of monocyte chemotactic protein 1 (MCP-1) and interleukin-8 (IL-8) and the enzyme activities of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) were detected by ELISA assays.The mRNA expression of inducible nitric oxide synthase (iNOS) and COX-2 genes were detected by Real-time quantitative PCR,and the protein expression of iNOS and COX-2 were detected by Western blotting.The results showed that the secretions of MCP-1,IL-8 and the activities of intracellular COX-1 and COX-2 were extremely significantly higher after PCV2 infection than those in the blank control group,the mRNA expression levels and protein expression of intracellular iNOS and COX-2 were also extremely significantly increased (P<0.01);After treatment with SSP at 100,200 and 400 μg/mL,the secretions of MCP-1,IL-8 and the activities of COX-1 and COX-2 in the cells were significantly or extremely significantly decreased (P<0.05,P<0.01),the effect was most significant in 400 μg/mL SSP group,and the mRNA expression of intracellular iNOS and COX-2 gene was also significantly or extremely significantly decreased(P<0.05,P<0.01).The increase of iNOS protein expression induced by PCV2 was significantly or extremely significantly inhibited by treatment with SSP at 100,200 and 400 μg/mL (P<0.05,P<0.01).The increase of COX-2 protein expression was significantly inhibited by treatment with SSP at 200 and 400 μg/mL,and the best effect was obtained at 400 μg/mL.The results showed that SSP could inhibit the increase of mRNA expression and protein expression of pro-inflammatory factors caused by PCV2 infection and played a certain role in alleviating inflammation.

Key words: Sophora subprosrate polysaccharide; PCV2; 3D4/2; inflammatory factors

中图分类号:

S859.7

贾妮娜, 曹迷霞, 杨剑, 刘梦倩, 陈奇, 韦英益, 胡庭俊. 山豆根多糖对猪圆环病毒Ⅱ型感染猪肺泡巨噬细胞炎性因子水平的影响[J]. 中国畜牧兽医, 2021, 48(5): 1825-1831.

JIA Nina, CAO Mixia, YANG Jian, LIU Mengqian, CHEN Qi, WEI Yingyi, HU Tingjun. Effect of Sophora subprosrate Polysaccharide on the Levels of Inflammatory Factors in 3D4/2 Cells Infected with Porcine Circovirus Virus Ⅱ Type[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(5): 1825-1831.

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