| 肝脏特异性Nampt基因敲除对缺血性脑卒中的影响 |
| 投稿时间：2021-07-22 修订日期：2021-10-27 点此下载全文 |
| 引用本文：青胜利,王淑娜,汪东昇,吕小群,徐添颖,缪朝玉.肝脏特异性Nampt基因敲除对缺血性脑卒中的影响[J].药学实践杂志,2022,40(1):12~19 |
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| 基金项目:国家自然科学基金资助项目(81730098); 上海市科委动物专项(16140904500) |
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| 中文摘要:目的 烟酰胺磷酸核糖转移酶(nicotinamide phosphoribosyltransferase，Nampt)是缺血性脑卒中的治疗新靶点。本研究旨在阐明肝脏来源的Nampt是否对缺血性脑卒中具有保护作用。方法 运用Cre/loxP系统制备肝脏特异性Nampt基因敲除小鼠。将NamptloxP/loxP小鼠与肝脏特异性表达Cre重组酶小鼠(Alb-Cre)进行杂交，采用聚合酶链反应方法鉴定子代基因型。测定基因敲除小鼠和同窝对照小鼠的体重。蛋白免疫印迹法检测小鼠肝脏和脑中Nampt蛋白的表达。采用电凝法对肝脏特异性Nampt基因敲除小鼠和对照小鼠制备大脑中动脉阻塞(middle cerebral artery occlusion，MCAO)脑卒中模型，造模24 h后对各组小鼠进行神经功能损伤评分，TTC染色测定脑梗死体积，ELISA法检测各组小鼠血浆Nampt水平。结果 成功构建肝脏特异性Nampt基因敲除小鼠，其基因型为NamptloxP/loxP Alb-Cre。肝脏特异性Nampt基因敲除组肝脏Nampt蛋白表达与对照组相比下降74.2%。脑Nampt蛋白的表达在敲除组与对照组之间无显著性差异。肝脏特异性Nampt基因敲除对小鼠的体重无影响。正常生理条件下，同性别肝脏特异性Nampt基因敲除小鼠与对照小鼠血浆Nampt水平无明显差异。MCAO造模24 h后，肝脏特异性Nampt基因敲除组与对照组神经行为学损伤评分、脑梗死体积和血浆Nampt浓度也无显著性差异。结论 成功构建肝脏特异性Nampt基因敲除小鼠；肝脏来源Nampt对缺血性脑卒中没有明显保护作用。 |
| 中文关键词:烟酰胺磷酸核糖转移酶 特异性敲除 肝脏 脑卒中 |
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| Effects of liver-specific Nampt knockout on ischemic stroke |
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| Abstract:Objective Nicotinamide phosphoribosyltransferase (Nampt) is a new therapeutic target for ischemic stroke. The aim of this study was to investigate protective effect of liver-derived Nampt on ischemic stroke. Methods Liver-specific Nampt knockout mice were generated using the Cre/loxP system. NamptloxP/loxP mice were crossed with liver-specific Cre recombinase expression mice (Alb-Cre), and the progeny genotypes were identified by polymerase chain reaction. Body weight of knockout mice and control mice were measured. Nampt in liver and brain was determined by Western blot assay. Middle cerebral artery occlusion (MCAO), a classical ischemic stroke model, was generated in liver-specific Nampt knockout mice and control mice by electrocoagulation. After 24 h of modeling, neurological deficit scores of each group were evaluated and TTC staining was performed to determine the cerebral infarction volume. The level of plasma Nampt in each group was determined by ELISA. Results Liver-specific Nampt knockout mice with the genotype of NamptloxP/loxPAlb-Cre were successfully constructed. The hepatic Nampt expression in knockout mice was significantly decreased by 74.2% compared to control mice, while there was no significant difference in the expression of brain Nampt protein between the knockout group and the control group. Specific knockout of liver Nampt gene expression had no effect on the body weight of mice. Under normal physiological conditions, there was no significant difference in plasma Nampt levels between liver-specific Nampt knockout mice and control mice of the same gender. 24 h after MCAO modeling, there were no significant differences in neurological deficit scores, cerebral infarct volume and plasma Nampt concentration between liver-specific Nampt knockout group and control group. Conclusion Liver-specific Nampt knockout mice are successfully constructed. Liver-derived Nampt has no significant protective effects on ischemic stroke. |
| keywords:Nampt specific knockout liver stroke |
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