姜黄素对玉米赤霉烯酮致小鼠肝脏氧化应激和NLRP3炎症小体活化的影响

Effects of curcumin on oxidative stress and NLRP3inflammasome activation induced by zearalenone in mouse liver

本研究旨在探究姜黄素对玉米赤霉烯酮(zearalenone,ZEA)诱导小鼠的肝脏氧化应激和NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体活化的影响。60只雄性健康昆明小鼠随机分为对照组、姜黄素组(150mg/kg)、ZEA组(40mg/kg)、姜黄素+ZEA组。灌胃给药4周后收集小鼠血清和肝脏样品,测定血清中谷草转氨酶(AST)和谷丙转氨酶(ALT)活性;HE染色与透射电镜分别观察肝脏组织病理形态改变和超微结构;二氢乙啶(DHE)染色测定肝脏组织活性氧(ROS)水平;比色法检测谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)和超氧化物歧化酶(SOD)活性以及丙二醛(MDA)含量;免疫荧光检测NLRP3和半胱氨酸蛋白酶-1(Caspase-1)p20蛋白表达;荧光定量PCR和酶联免疫吸附实验(ELISA)测定白细胞介素1β(IL-1β)水平。结果表明,与对照组相比,ZEA组小鼠肝脏发生明显病理学变化,肝细胞线粒体损伤较重。ZEA组小鼠血清AST和ALT活性升高,肝脏ROS水平和MDA含量增加,GSH-Px、CAT和SOD活性降低,较对照组差异显著(P<0.01)。与ZEA组相比,姜黄素+ZEA组小鼠肝脏病理学变化缓解,线粒体损伤程度改善,血清AST和ALT活性降低(P<0.01),肝组织中ROS水平和MDA含量降低(P<0.01),GSH-Px、CAT和SOD活性提高(P<0.01)。与ZEA组相比,姜黄素干预降低了ZEA诱导的肝脏NLRP3和Caspase-1p20蛋白表达以及IL-1β水平升高(P<0.01)。综上所述,姜黄素对ZEA诱导的小鼠肝脏损伤有保护作用,可能与姜黄素发挥抗氧化和抗炎作用有关。

The aim of this experiment was to explore the effects of curcumin on zearalenone(ZEA)-induced oxidative stress and nod-like receptor family pyrin domain containing 3(NLRP3)activation in mice liver.Sixty healthy male Kunming mice were randomly divided into control group,curcumin group(150mg/kg),ZEA group(40mg/kg),and curcumin+ZEA group.The serum and liver samples were harvested after 4weeks of intragastric administration.The serum aspartate aminotransferase(AST)and alanine transaminase(ALT)activities were measured.The pathological morphology changes and ultrastructure of livers were observed by HE staining and transmission electron microscopy,respectively.The reactive oxygen species(ROS)levels in liver tissues were detected by dihydroethidine(DHE)staining.The activities of glutathione peroxidase(GSH-Px),catalase(CAT),and superoxide dismutase(SOD),and the content of malonaldehyde(MDA)were measured by colorimetry.The proteins expressions of NLRP3and Caspase-1p20were detected by immunofluorescence.The levels of interleukin-1β(IL-1β)were determined by real-time quantitative PCR and enzyme-related immunosorbent assay(ELISA).The results indicated that the livers from ZEA group had obvious pathological changes and the mitochondrial damage was more serious as compared with control group.The activities of serum AST and ALT in ZEA group were increased,the levels of ROS and MDA in liver were increased,and the activities of GSH-Px,CAT and SOD were decreased,which were significantly different from those in control group(P<0.01).Compared with ZEA group,the pathological changes of livers in curcumin+ZEA group were relieved,the degree of mitochondrial damage was improved,the activities of AST and ALT in serum were decreased(P<0.01),the levels of ROS and MDA in liver tissue were decreased(P<0.01),and the activities of GSH-Px,CAT and SOD were increased(P<0.01).Compared with ZEA group,curcumin intervention decreased the upregulation of NLRP3and Caspase-1p20protein expressions and IL-1βlevel induced by ZEA in liver(P<0.01).Taken together,the protective effect of curcumin on ZEA-induced liver damage in mice,which may be related to the anti-oxidant and anti-inflammatory effects of curcumin.