检索规则说明:AND代表“并且”;OR代表“或者”;NOT代表“不包含”;(注意必须大写,运算符两边需空一格)
检 索 范 例 :范例一: (K=图书馆学 OR K=情报学) AND A=范并思 范例二:J=计算机应用与软件 AND (U=C++ OR U=Basic) NOT M=Visual
作 者:熊款款 吴映欣 易思亮 杨凌宸[1] XIONG Kuankuan;WU Yingxin;YI Siliang;YANG Lingchen(College of Veterinary Medicine,Hunan Agricultural University,Changsha 410128,China)
出 处:《中国畜牧兽医》2021年第12期4710-4717,共8页China Animal Husbandry & Veterinary Medicine
基 金:国家自然科学基金青年基金项目(31802252);湖南省教育厅优秀青年项目(18B113)。
摘 要:试验旨在研究T-2毒素对猪肾细胞(PK15)的毒性作用及其氧化应激反应。用不同浓度的T-2毒素处理细胞,通过MTT法检测细胞活性、测定细胞内乳酸脱氢酶(LDH)释放率及苏木素伊红(HE)染色观察细胞形态变化评估T-2毒素对细胞的毒性作用;通过检测细胞内的活性氧(ROS)水平、丙二醛(MDA)含量及谷胱甘肽(GSH)含量、谷胱甘肽过氧化物酶(GPx)活性,评估不同剂量T-2毒素对细胞氧化应激水平的影响;检测Nrf2-ARE信号通路相关基因Nrf2、Keap1、GPx-1、Nqo1及Hmox1 mRNA的表达水平,分析T-2毒素对该信号通路的影响。结果表明,PK15细胞活性呈毒素剂量依赖性下降(P<0.05),LDH释放率呈毒素剂量依赖性上调(P<0.05);随T-2毒素剂量升高,细胞间隙逐渐增加,细胞固缩,细胞数量也随之减少。细胞内ROS阳性细胞率呈剂量依赖性升高,且T-2毒素为100 nmol/L时,ROS阳性细胞率显著高于其他剂量组(P<0.05);细胞内MDA含量与GPx活性呈毒素剂量依赖性升高(P<0.05),而GSH含量呈毒素剂量依赖性下降(P<0.05)。20、50及100 nmol/L的T-2毒素可显著上调Keap1、Gpx-1及Nqo1基因mRNA的相对表达量(P<0.05),且50 nmol/L的相对表达量最高。T-2毒素对K15细胞有毒性作用,可诱导其氧化损伤,且该氧化应激过程受Nrf2-AER信号通路调控。The purpose of this test was to study the toxic effect of T-2 toxin on porcine kidney cells(PK15)and its oxidative stress response.The cells were treated with different concentrations of T-2 toxin.The cytotoxicity of the toxin was evaluated by MTT assay,intracellular lactate dehydrogenase(LDH)release rate and HE staining.The effects of different doses of T-2 toxin on the level of oxidative stress were evaluated by detecting the levels of reactive oxygen species(ROS),malondialdehyde(MDA),total glutathione(GSH)and total glutathione peroxidase(GPx).The expression levels of Nrf2,Keap1,GPx-1,Nqo1 and Hmox1 genes mRNA related to Nrf2-ARE signal pathway were detected,and the effect of T-2 toxin on this signal pathway was analyzed.The results showed that the activity of PK15 cells decreased in a toxin dose-dependent manner(P<0.05),and the release rate of LDH increased in a toxin dose-dependent manner(P<0.05).With the increase of T-2 toxin dose,the cell gap gradually increased,the cell pyknosis and the number of cells decreased.The rate of ROS positive cells increased in a dose-dependent manner,and when T-2 toxin was 100 nmol/L,the rate of ROS positive cells was significantly higher than that in other dose groups(P<0.05).The intracellular MDA content and GPx activity increased in a toxin dose-dependent manner(P<0.05),while the GSH content decreased in a toxin dose-dependent manner(P<0.05).20,50 and 100 nmol/L T-2 toxin could significantly up regulate the relative expression of Keap1,GPx-1 and Nqo1 genes mRNA(P<0.05),and the relative expression of 50 nmol/L was the highest.T-2 toxin caused toxic effects on PK15 cells and induced oxidative damage,and the oxidative stress process was regulated by Nrf2-AER signal pathway.
关 键 词:T-2毒素 PK15细胞 毒性作用 氧化应激 Nrf2-ARE通路
分 类 号:S859.87[农业科学—临床兽医学]
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在链接到云南高校图书馆文献保障联盟下载...
云南高校图书馆联盟文献共享服务平台 版权所有©
您的IP:216.73.216.13