辣蓼黄酮正丁醇部位对PRV感染RAW264.7细胞氧化应激的干预作用及组蛋白乙酰化水平的影响

Effect of N-butanol fraction of Polygonum hydropiper flavonoids on oxidative stress and histone acetylation in PRV-infected RAW264.7 cells

试验旨在探究辣蓼黄酮正丁醇部位(FNB)对猪伪狂犬病毒(PRV)感染小鼠单核巨噬细胞(RAW264.7)氧化应激反应的干预作用及组蛋白乙酰化水平的影响。采用CCK-8法检测FNB对细胞的毒性作用,采用不同浓度的FNB(25、50、100 mg/L)处理PRV感染的RAW264.7细胞4、8、12 h,测定氧化应激相关因子和组蛋白乙酰化水平。结果显示,与PRV感染组相比,25 mg/L FNB作用于PRV感染的RAW264.7细胞12 h后,细胞内诱导性一氧化氮合酶(iNOS)活性及一氧化氮(NO)、丙二醛(MDA)水平降低,谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、小鼠血红素氧合酶1(HO-1)和小鼠醌氧化还原酶1(NQO1)活性升高,iNOS mRNA的表达水平极显著下调(P<0.01),HO-1 mRNA的表达水平显著上调(P<0.05),核因子E2相关因子2(Nrf2)、NQO1的表达水平极显著上调(P<0.01)。与PRV感染组相比,25 mg/L FNB组处理12 h时后,细胞内组蛋白乙酰化酶(HAT)的活性降低,组蛋白去乙酰化酶(HDAC)的活性升高,细胞内HAT mRNA的表达水平下调,HDAC mRNA的表达水平上调。研究表明,FNB对PRV感染RAW264.7细胞诱导的氧化应激具有一定的调节作用,可通过提高细胞内多种抗氧化酶的基因表达水平发挥抗氧化作用,调节细胞内的乙酰化水平,降低PRV感染导致的氧化应激损伤。

The study was carried out to investigate the effect of N-butanol fraction of Polygonumhydropiperflavonoids(FNB)on oxidative stress and histone acetylation in PRV-infected RAW264.7 cells.The effect of FNB on viability of the cells was detected by cell counting kit(CCK-8),and the levels of oxidative stress-related factors and histone acetylation were measured when the PRV infected cells were treated with FNB at 25,50,100 mg/L for 4,8,12 h.The results showed that compared to the prv-infected group,after 25 mg/L FNB was applied to PRV-infected RAW264.7 cells for 12 h,the activity of intracellular iNOS and the levels of NO and MDA decreased,and the activities of GSH-Px,SOD,CAT,mouse HO-1,and mouse NQO1 increased.The expression levels of iNOSmRNA were extremely down-regulated(P<0.01),the expression levels of HO-1 mRNA were significantly up-regulated(P<0.05),and the expression levels of nuclear factor E2-related factor 2(Nrf2)and NQO1 were extremely up-regulated(P<0.01).Compared with PRV-infected group,the activity of intracellular histone acetylase(HAT)decreased,histone deacetylase(HDAC)activity increased,intracellular HAT mRNA expression level was down-regulated,and HDAC mRNA expression level was up-regulated after 12 h of 25 mg/L FNB treatment.The results showed that FNB had a certain regulatory effect on the oxidative stress induced by PRV infection in RAW264.7 cells,and exerted antioxidant effects by increasing the gene expression levels of various antioxidant enzymes in cells,regulating the level of acetylation in cells,and reducing the oxidative stress damage caused by PRV infection.