甘草多糖对鼠伤寒沙门菌诱导的巨噬细胞氧化-抗氧化平衡紊乱的调解  

Effect of radix glycyrrhiza polysaccharide on the imbalance of oxidation/antioxidation induced by Salmonella Typhimuriumin murine peritoneal macrophages

作　　者：廖成水[1] 王晓利 牛晓婉 邵文超 王楠[1] 孙召金[1] 程相朝[3] 刘明远[1] 

机构地区：[1]吉林大学人兽共患病研究所人兽共患病研究教育部重点实验室,吉林长春130062 [2]汝州市畜牧局,河南汝州467500 [3]河南科技大学动物科技学院,河南洛阳471003

出　　处：《中国兽医学报》2015年第6期941-945,共5页Chinese Journal of Veterinary Science

摘　　要：为了探讨甘草多糖对鼠伤寒沙门菌诱发巨噬细胞氧化-抗氧化平衡紊乱的调节作用。将0、1、20和100 mg/L甘草多糖4个质量浓度组与小鼠腹腔巨噬细胞共孵育,同时每组按1∶1加入鼠伤寒沙门菌SL1344;利用ELISA试剂盒检测各组丙二醛(malondialdehyde,MDA),活性氧(reactive oxygen species,ROS),一氧化氮(nitricoxide,NO),诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS),谷胱甘肽过氧化物酶(glutathione peroxidase,GSHPX),超氧化物歧化酶(superoxide dismutase,SOD)和总抗氧化能力(total antioxidant capacity,T-AOC)变化情况;利用荧光显微镜和荧光酶标仪观察甘草多糖对鼠伤寒沙门菌引起巨噬细胞产生ROS的影响;利用乳酸脱氢酶(lactate dehydrogenase,LDH)细胞毒性试剂盒检测巨噬细胞上清LDH水平。结果表明,鼠伤寒沙门菌可诱导小鼠腹腔巨噬细胞发生氧化损伤,导致NO(4.65±1.07)μmol/L、iNOS(15.46±0.92)U/mL、MDA(3.46±0.39)nmol/L和ROS(3.69±0.31)U/mL氧化水平以及LDH水平显著升高,而GSH-PX(15.58±1.35)nmol/L、SOD(13.32±0.92)NU/mL和T-AOC(8.36±0.60)U/mL抗氧化水平显著降低。荧光显微镜和荧光酶标仪检测显示,鼠伤寒沙门菌感染小鼠腹腔巨噬细胞ROS水平显著升高。甘草多糖呈剂量依赖明显降低细胞氧化水平和增强抗氧化水平。甘草多糖作为重要的免疫调节剂,能够调节鼠伤寒沙门菌引起的巨噬细胞氧化损伤,维持细胞氧化-抗氧化平衡。To determine the effect of radix glycyrrhiza polysacchride on the regulation of the imbalance of oxidation/antioxidation induced by Salmonella Typhimuriumin murine peritoneal macrophages.Radix glycyrrhiza polysacchride(0,1,20 and 100mg/L)was applied onto macrophage for3 h.Salmonella typhimurium SL1344 was added to the cultured cells at a multiplicity of infection(MOI)of 1∶1.The level of malondialdehyde(MDA),reactive oxygen species(ROS),nitricoxide(NO),inducible nitric oxide synthase(iNOS),glutathione peroxidase(GSH-PX),superoxide dismutase(SOD),and total antioxidant capacity(T-AOC)were measured by ELISA.Fluorescence microscopy and fluorescence microplate reader were used to detect measure the ROS levels of macrophages.The LDH activity in supernatants was determined using a LDH Cytotoxicity Assay Kit.The results indicated that oxidative damage of macrophages exposed to Salmonella Typhimurium was observed.A dramatically higher the level of NO(4.65±1.07)μmol/L,iNOS(15.46±0.92)U/mL,MDA(3.46±0.39)nmol/L,ROS(3.69±0.31)U/mL and LDH,decreased the level of GSH-PX(15.58±1.35)nmol/L,SOD(13.32±0.92)NU/mL and T-AOC(8.36±0.60)U/mL was found in drugs group than that of control group.Radix Glycyrrhiza polysacchride(20and 100mg/L)was able to reduce the levels of MDA,ROS and LDH,and improve the levels of NO,iNOS,GSH-PX,SOD and T-AOC in a dose dependent manner.The data indicated radix glycyrrhiza polysacchride is effective for improving the imbalance of oxidation/antioxidation of murine peritoneal macrophages-infected with Salmonella Typhimurium.

关 键 词：甘草多糖 巨噬细胞 氧化应激 鼠伤寒沙门菌 

分 类 号：S853.74[农业科学—临床兽医学]