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严森森,吕梦迪,杨学思,陶映君,徐光翠,赵英政.硫辛酸合成酶低表达对小鼠抗氧化能力的影响[J].中国实验动物学报,2022,30(1):64~69.
硫辛酸合成酶低表达对小鼠抗氧化能力的影响
Effects of low expression of lipoic acid synthase on antioxidant capacity in mice
投稿时间:2021-08-30  
DOI:10. 3969 / j.issn.1005-4847. 2022. 01. 008
中文关键词:  硫辛酸合成酶  基因分型  氧化损伤  小鼠模型
英文关键词:lipoic acid synthase  genotyping  oxidative damage  mice model
基金项目:
作者单位E-mail
严森森 新乡医学院公共卫生学院,河南 新乡 453003 xhldyjs@ 163.com 
吕梦迪 新乡医学院公共卫生学院,河南 新乡 453003  
杨学思 新乡医学院公共卫生学院,河南 新乡 453003  
陶映君 新乡医学院公共卫生学院,河南 新乡 453003  
徐光翠 新乡医学院公共卫生学院,河南 新乡 453003  
赵英政 新乡医学院公共卫生学院,河南 新乡 453003 yingzhengzhao@ 126.com 
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中文摘要:
       目的 建立一种硫辛酸合成酶基因低表达的基因修饰小鼠,并检测其抗氧化能力。 方法 将硫辛酸合成酶基因 3’-UTR 区域 Loxp 小鼠与 E2a-Cre 小鼠杂交,从子代小鼠中鉴定出 LiasL/ +小鼠并进一步繁殖、鉴定基因型。 选取基因型为 LiasL/ L和 Lias+ / +的 8 周龄雄性小鼠各 10 只;称重、麻醉后经股动脉采血,然后分离小鼠肺、肾及肝,并称重。应用 Western Blot、免疫组化方法检测肺、肾、肝组织中硫辛酸合成酶(LIAS)蛋白的表达水平;分离血清,检测总抗氧化能力(TAC);对肾、肝、肺匀浆后提取总蛋白,检测其 SOD、CAT 酶活性及 MDA 含量。 结果 Western Blot 及免疫组化分析显示,除小鼠肝 LIAS 免疫组化分析结果外,LiasL/ L组小鼠较 Lias+ / +组小鼠肾、肝及肺组织中 LIAS 蛋白表达水平均显著降低(P< 0. 05)。 与 Lias+ / +组小鼠相比,LiasL/ L组小鼠血清 TAC 水平显著降低(P< 0. 05),LiasL/ L小鼠肾、肝、肺脏器中 SOD、CAT 酶活性显著降低,MDA 水平显著升高(P< 0. 05)。 结论 本研究鉴定了硫辛酸合成酶基因低表达的基因修饰小鼠,其在肾、肝及肺中表达较野生型小鼠均降低,并导致相应脏器的氧化损伤。
英文摘要:
       Objective To establish a lipoic acid synthase gene-modified mouse model and detect their antioxidant capacity. Methods Mice with a loxP-modified 3′-UTR region of the lipoic acid synthase gene were hybridized with EIIa- Cre mice. LiasL/ + mice were identified from their offspring genotypes, and further propagated. Three genotypes of offspring were identified. Ten 8-week-old male mice with LiasL/ L and Lias+ / + genotypes were randomly selected. After weighing and anesthesia, blood was collected via the femoral artery, and then the kidneys, livers and lungs were collected and weighed. Western Blot and immunohistochemistry were used to detect expression levels of lipoic acid synthase (LIAS) protein in kidney, liver and lung tissue. Serum was collected to detect total antioxidant capacity (TAC). The total protein of tissue homogenates from kidney, liver and lung samples were used to detect the activities of superoxide dismutase and catalase, and the content of malondialdehyde. Results The expression levels of Lias in kidney, liver and lung samples from LiasL/ L mice were significantly lower than those from Lias+ / + mice by semi-quantitative western blotting and immunohistochemical analysis (P< 0. 05). Compared with the Lias+ / + group, serum levels of TAC and enzyme activities of superoxide dismutase and catalase in kidney, liver and lung samples were decreased, and the malondialdehyde level increased in the LiasL/ L group (P< 0. 05). Conclusions Kidney, liver and lung expression levels of Lias were lower in mice with a modified 3′- UTR region of Lias than those of wild-type mice, with corresponding oxidative damage observed in these organs.
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