星形胶质细胞敲除铜蓝蛋白对小鼠脑铁代谢的影响  

Effects of conditional knockout ceruloplasmin in astrocytes on brain iron metabolism of mice

作　　者：李海艳[1,2] 李忠达 何立娟 LI Hai-yan;LI Zhong-da;HE Li-juan(Department of Human Anatomy,Chengde Medical College,Hebei Chengde067000,China;The Key Laboratory of Animal Physiology,Biochemistry and Molecular Biology of Hebei Province,College of Life Sciences,Hebei Normal University,Shijiazhuang050024,China)

机构地区：[1]承德医学院人体解剖学教研室,河北承德067000 [2]河北师范大学生命科学学院河北省动物生理生化与分子生物学重点实验室,石家庄050024

出　　处：《解剖学报》2024年第4期475-481,共7页Acta Anatomica Sinica

基　　金：承德医学院高层次人才科研启动基金(202203);承德医学院人体解剖学与组织学胚胎学优势学科资助项目(202302)。

摘　　要：目的探讨铜蓝蛋白(CP)在脑铁代谢中的作用机制。方法将CP^(flox/flox)小鼠与^(GFAP)-cre小鼠杂交,得到胶质纤维酸性蛋白(^(GFAP))-cre介导的CP基因敲除小鼠,即CP^(GFAP)cKO小鼠。6月龄88只雄性小鼠被分为WT、^(GFAP)-cre、CP^(flox/flox)和CP^(GFAP)cKO 4组。通过基因型鉴定、免疫荧光染色和Western blotting方法检测星形胶质细胞是否特异性敲除CP;Morris水迷宫实验检测星形胶质细胞敲除CP的小鼠学习和记忆功能;通过电感耦合等离子体质谱法(ICP-MS)和同步辐射微束X线荧光光谱法(μ-XRF)检测皮层、海马区铁水平;血清和组织铁试剂盒测定血清和肝铁含量。结果基因型鉴定、免疫荧光染色和Western blotting结果确认星形胶质细胞特异性敲除CP基因小鼠建立模型成功;Moris水迷宫实验结果观察到星形胶质细胞特异性敲除CP基因,小鼠的学习记忆能力明显降低;ICP-MS、μ-XRF结果显示,皮层区和海马区铁含量显著下降;而血清铁和肝铁含量没有明显变化。结论星形胶质细胞特异性敲除CP基因的小鼠的学习记忆能力下降与脑铁降低密切相关;CP的主要功能可能是协助脑细胞摄取铁。Objective To explore the mechanism of ceruloplasmin(CP)in brain iron metabolism.Methods We obtained glial fibrillary acidic protein(^(GFAP))-cre-mediated ceruloplasmin knockout mice(CP^(GFAP)cKO mice)by crossing CP^(flox/flox)mice with transgenic^(GFAP)-cre mice.Eighty-eight male mice aged 6 months were divided into four groups,WT,^(GFAP)-cre,CP^(flox/flox)and CP^(GFAP)cKO.To observe whether astrocytic CP gene knockout specifically through genotype identification and immunofluorescent staining,and Western blotting method.The learning and memory function was detected by Morris water maze,brain iron levels in the cortex and hippocampus of mice were measured by inductively coupled plas mamass spectrometry(ICP-MS)andμ-probe X-ray fluorescence(μ-XRF),and iron contents in serum and liver were detected by iron kit.Results The mice of conditional knockout ceruloplasmin in astrocytes were obtained successfully by genotype identification,immunofluorescent staining and Western blotting method.The learning and memory ability was reduced in the water maze experiment,and iron contents decreased in the cortex and hippocampal regions of 6-month-old CP^(GFAP)cKO group.While the level of serum iron and liver iron in the four groups did not change significantly.Conclusion The decline of learning and memory ability of mice with astrocytic CP gene knockout specifically is closely related to the decrease in brain iron.The main function of CP may be to help brain cells to absorb iron.

关 键 词：铜蓝蛋白 星形胶质细胞 铁 认知 免疫印迹法 小鼠 

分 类 号：Q421[生物学—神经生物学]