硒代蛋氨酸羟基类似物对H_(2)O_(2)诱导的鸡肝癌细胞氧化损伤的保护作用  

Protective effect of selenomethionine hydroxyl analogues on H_(2)O_(2)-induced oxidative damage in LMH cells

作　　者：刘永仕 吕茜 袁何玲 龚倩梅 王晓明 黄金虎[1] 王丽平[1] LIU Yongshi;LYU Xi;YUAN Heling;GONG Qianmei;WANG Xiaoming;HUANG Jinhu;WANG Liping(College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学动物医学院,江苏南京210095

出　　处：《畜牧与兽医》2024年第5期35-43,共9页Animal Husbandry & Veterinary Medicine

基　　金：江苏省自主创新项目[CX(22)3039];江苏省研究生科研与实践创新计划项目(SJCX21_0239)。

摘　　要：旨在探索硒代蛋氨酸羟基类似物(SeOH)缓解鸡肝癌细胞(LMH)氧化损伤的作用及潜在机制。建立H_(2)O_(2)氧化应激模型,通过检测细胞活力和细胞氧化/抗氧化指标的变化以及核转录因子E2相关因子2(Nrf_(2))通路和内质网应激关键分子的表达情况,评价不同浓度SeOH(16、32和64μg/mL)的抗氧化应激和内质网应激作用。结果显示:120μmol/L的H_(2)O_(2)刺激细胞24 h时,细胞活力显著下降(P<0.001),活性氧(ROS)的荧光强度升高。不同剂量的SeOH(16、32和64μg/mL)均可显著提高H_(2)O_(2)处理后的LMH细胞活力(P<0.01);32和64μg/mL的SeOH均可极显著提高LMH细胞的超氧化物歧化酶(SOD)活性(P<0.01),显著降低丙二醛(MDA)含量(P<0.001)以及ROS的荧光强度。进一步研究显示32和64μg/mL的SeOH极显著上调Nrf_(2)蛋白(P<0.01)并显著上调谷胱甘肽过氧化物酶4(GPX4)蛋白的表达水平(P<0.05),极显著下调Kelch样ECH相关蛋白1(Keap1)蛋白的表达水平(P<0.001),极显著增加Nrf_(2)的入核率(P<0.001);64μg/mL的SeOH可显著降低葡萄糖调节蛋白78(GRP78)(P<0.05)、蛋白激酶RNA样内质网激酶(PERK)(P<0.01)和转录激活因子4(ATF4)(P<0.001)的表达水平。提示:SeOH能通过激活Nrf_(2)通路并且抑制内质网应激缓解H_(2)O_(2)诱导的LMH细胞氧化应激,为畜禽养殖中SeOH作为抗氧化剂的应用提供理论基础。The aim of this study was to explore the effect and potential mechanism of hydroxy-selenomethionine(SeOH)on oxidative dam-age of chicken hepatocellular carcinoma cells(LMH).An H_(2)O_(2) oxidative stress model was established to evaluate the effects of different con-centrations of SeOH(16,32 and 64μg/mL)on oxidative stress and ER stress by detecting the changes in cell viability and cell oxidation/antioxidant indexes,as well as the expression of the nuclear factor erythroid-related factor 2(Nrf_(2))pathway and the key molecules of ER stress.The results showed that cell viability significantly decreased(P<0.001)and the fluorescence intensity of ROS increased,indicating that the H_(2)O_(2)-induced(120μmol/L,for 24 h)oxidative stress model was successfully established.The different concentrations of SeOH(16,32 and 64μg/mL)significantly enhanced the cell viability(P<0.01).Both 32 and 64μg/mL SeOH significantly promoted the activi-ty of superoxide dismutase(SOD)in the LMH cells(P<0.01),significantly reduced the content of malondialdehyde(MDA)(P<0.001)and the production of fluorescence intensity of ROS in the LMH cells.Further research showed that SeOH(32 and 64μg/mL)significantly up-regulated the protein expression level of Nrf_(2)(P<0.01)and glutathione peroxidase 4(GPX4)(P<0.05),and down-regulated the ex-pression level of Kelch-like ECH-associated protein 1(Keap1)(P<0.001),and synergistically promoted the efficiency of nuclear translo-cation of Nrf_(2)(P<0.001).However,the protein expression levels of GRP78(P<0.05),PERK(P<0.01)and ATF4(P<0.001)were significantly decreased under the 64μg/mL SeOH treatment.In conclusion,SeOH relieved H_(2)O_(2)-induced oxidative stress in LMH cells by activating the Nrf_(2) pathway and inhibiting ER stress.This study provided a theoretical basis for the application of SeOH as an antioxidant in livestock and poultry industries.

关 键 词：SeOH 氧化应激 Nrf_(2)信号通路 内质网应激 

分 类 号：S859[农业科学—临床兽医学]