【摘要】 目的:反式脂肪酸(trans-fatty acid,TFA)主要来源于食品工业生产过程，TFA危害健康是全球关注的公共卫生问题。亚麻木酚素(secoisolariciresinol diglucoside,SDG)是植物多酚类化合物，具有抗氧化性能。本研究旨在探讨SDG对妊娠期及哺乳期母代TFA暴露致子代小鼠肾氧化损伤的保护作用。方法:30只c57BL/6雌鼠随机分为5组，分别为对照组、TFA暴露组及SDG低(TFA+LSDG)、中(TFA+MSDG)、高(TFA+HSDG)剂量干预组，每组6只。除对照组外，其余4组母鼠实验期间每日经口灌胃TFA 1次，剂量为60 mg/(kg·BW)；对照组母鼠用生理盐水每日灌胃1次；同时3组SDG干预组分别用含10、20、30 mg/kg SDG饲料自由喂养；隔夜受孕，第2天早晨出现阴栓判定为受孕并将其纳入实验阶段，至哺乳期第21天实验结束。记录子鼠体重、肾质量，计算肾脏系数；采用HE染色观察子鼠肾病理学变化；二氢乙啶(dihydroethidium,DHE)荧光探测针染色检测活性氧(reactive oxidative species,ROS)水平；子鼠肾组织匀浆后检测总超氧化物歧化酶(total Superoxide Dismutase,T-SOD)活性和丙二醛(malondialdehyde,MDA)含量；采用免疫组织化学(immunohistochemistry,IHC)染色分析核因子E2相关因子2(nuclear factor E2-related factor 2,Nrf2)和血红素加氧酶-1(hemeoxygenase-1,HO-1)蛋白的表达；real-time PCR检测Nrf2和HO-1 mRNA表达；蛋白质印迹法检测抗氧化酶系铜锌超氧化物歧化酶(Cu/Zn-superoxide dismutase,Cu/Zn-SOD)、锰超氧化物歧化酶(Mn-superoxide dismutase,Mn-SOD)、谷胱甘肽过氧化物酶1(glutathione peroxidase-1,Gpx-1)蛋白的表达和通路相关蛋白Nrf2、HO-1的表达。结果:与对照组相比，TFA暴露组子鼠肾脏系数上升；肾组织形态结构异常；T-SOD酶活性下降，MDA含量上升；ROS水平升高；抗氧化酶系Cu/Zn-SOD、Mn-SOD、GPx-1蛋白的表达水平下降；Nrf2、HO-1 mRNA表达下降，且Nrf2和HO-1蛋白的表达水平下降，差异均有统计学意义(均P<0.05)。与TFA暴露组相比，SDG低、中、高剂量组均出现ROS水平降低，T-SOD酶活性、Cu/Zn-SOD、GPx-1、Mn-SOD、Nrf2和HO-1蛋白的表达水平上调；中、高剂量组可致子鼠肾脏系数及MDA含量下降；高剂量组可上调Nrf2 mRNA表达，差异均有统计学意义(均P<0.05)。结论:妊娠期及哺乳期母鼠TFA暴露会导致子代肾氧化损伤，而SDG干预可能通过Nrf2/HO-1通路来减轻TFA诱导的氧化损伤。更多还原

【Abstract】 Objective: Trans-fatty acids(TFAs), primarily derived from the food industry’s production processes, have become a globally recognized public health issue due to the detrimental impact they have on human well-being. Secoisolariciresinol diglucoside(SDG) is a polyphenolic compound derived from flax lignans, possessing antioxidative properties. This study aims to investigate the protective effect of SDG on kidney oxidative damage in offspring of mice caused by maternal exposure to TFA during pregnancy and lactation.Methods: A total of 30 c57BL/6 female rats were randomly divided into 5 groups: a control group, a TFA-exposed group, a low-(TFA+LSDG) group, a medium-(TFA+MSDG) group, and a high-(TFA+HSDG) group(n=6 in each group). With the exception of the control group, the maternal mice in the remaining 4 groups received a daily oral gavage of TFA at a dosage of 60 mg/(kg·BW) throughout the experimental period. The mothers in the control group were administered physiological saline via oral gavage once daily. Meanwhile, the 3 SDG intervention groups were provided with ad libitum access to SDG feed containing 10 mg/kg(low), 20 mg/kg(medium), and 30 mg/kg(high) of SDG. The female mice were conceived overnight. If the vaginal plug appeared in the next morning, the female mice were conceived and included in the experimental stage until the end of the 21th day lactation period. The body weight and kidney mass of offspring were recorded, and the kidney coefficient was calculated. The kidney was detected by HE staining to observe the histopathological changes, and the level of reactive oxidative species(ROS) was detected by fluorescence probe-dihydroethidium(DHE) staining; the expression levels of total superoxide dismutase(T-SOD) and malondialdehyde(MDA) in renal homogenate were detected, and the expression of nuclear factor E2-related fator2(Nrf2) and hemeoxygenase-1(HO-1) protein was analyzed by immunohistochemistry(IHC) staining. The mRNA expressions of Nrf2 and HO-1 were detected by real-time PCR, and the protein expression of Cu/Zn-superoxide dismutase(Cu/Zn-SOD), Mn-superoxide dismutase(MnSOD), glutathione peroxidase-1(GPx-1), Nrf2 and HO-1 were detected by Western blotting.Results: Compared with the control group, the kidney coefficient in the TFA-exposed group was increased, the morphology and structure of kidney tissue was abnormal; the activity of T-SOD enzyme was decreased, and the content of MDA was increased, the level of ROS was increased; the expressions of Cu/Zn-SOD, Mn-SOD, GPx1 protein were decreased, and the mRNA and protein expressions of Nrf2 and HO-1 were decreased, there were all significant difference(all P<0.05). Compared with the TFA-exposed group, the ROS levels were reduced, and the T-SOD enzyme activity as well as the protein expression of Cu/Zn-SOD, GPx-1, Mn-SOD, Nrf2 and HO-1 were up-regulated in the low, middle and high dose SDG intervention groups; the kidney coefficient and MDA content were decreased in the middle and high dose SDG groups; the Nrf2 mRNA expression in the high dose SDG group was up-regulated, there were all significant difference(all P<0.05).Conclusion: Maternal exposure to TFA during pregnancy and lactation can lead to oxidative damage in the kidney of offspring, and the SDG intervention may alleviate TFAinduced oxidative damage by up-regulating the expression of Nrf2 and HO-1 signal pathway.更多还原