妊娠期糖尿病致子鼠神经系统发育异常的分子学机制探讨及微RNA-7047-5p的验证  

Molecular mechanism and validation of miRNA-7047-5p for abnormal nervous system development in gestational diabetes mellitus in mice

作　　者：黄婉仪[1] 王丽娜[1] 欧巧群[1] 胡家奇 张又祥[1] HUANG Wanyi;WANG Lina;OU Qiaoqun;HU Jiaqi;ZHANG Youxiang(Department of Pediatrics,Guangzhou First people's Hospital,Guangzhou,Guangdong 510180,China)

机构地区：[1]广州市第一人民医院儿科,广东广州510180

出　　处：《安徽医药》2023年第10期1955-1959,I0004,共6页Anhui Medical and Pharmaceutical Journal

基　　金：广州市卫生健康科技西医类一般引导项目(20201A011003)。

摘　　要：目的探究妊娠期糖尿病(GDM)致子鼠神经系统发育异常的分子学机制及微RNA-7047-5p(miRNA-7047-5p)的表达验证。方法2020年5月至2021年12月,取30只昆明小鼠(雌性20只、雄性10只),以雄雌比例1∶2合笼过夜构建妊娠小鼠,采用随机数字表法将妊娠小鼠分为造模组和对照组,前者采用腹腔注射链脲佐菌素(STZ)方式构建GDM小鼠模型。血糖仪检测随机血糖;解剖显微镜观察子鼠脑组织整体结构;苏木精-伊红(HE)染色观察海马组织细胞形态;基因芯片实验检测子鼠海马组织差异表达微小核糖核酸(miRNA)基因;差异表达miRNA基因进行京都基因和基因组百科全书(KEGG)分析;定量聚合酶链式反应(q-PCR)检测miRNA-7047-5p表达。结果与对照组比,于给药后次日(E1.5)起造模组小鼠空腹血糖上升,E1.5、E5.5、E11.5、E18.5血糖值分别为(5.67±1.06)、(6.84±1.23)、(5.61±1.04)、(12.08±2.16)、(5.49±1.05)、(14.27±2.71)、(5.53±1.03)、(15.14±2.86)mmol/L(P<0.05);自给药后第1天起造模组小鼠随机血糖上升,给药后第1天、第2天、第3天、E4.5、E10.5、E17.5随机血糖分别为(5.83±1.06)、(22.36±3.74)、(5.79±1.03)、(20.65±3.49)、(5.95±1.11)、(19.94±2.97)、(5.93±1.14)、(18.05±3.14)、(5.87±1.15)、(16.49±2.85)、(5.69±1.03)、(14.89±2.37)mmol/L(P<0.05)。对照组子鼠脑组织整体无肿胀,脑沟和脑回清晰;GDM组子鼠脑组织脑沟和脑回变浅;对照组子鼠海马CA1区细胞排列紧密、有序,细胞形态饱满,核仁清晰,无核固缩;GDM组子鼠海马CA1区细胞数量明显减少,排列紊乱,细胞空泡样变化,核仁不清晰,呈大量核固缩状态;正常组子鼠和GDM组子鼠海马组织共有11个差异下调表达miRNA基因;差异表达miRNA基因多数富集在Wnt信号通路、2型糖尿病、突触囊泡循环、鞘脂代谢、加压素调节的水再吸收、催乳素信号通路、Hippo信号通路等通路;选定差异下调表达miRNA-7047-5p,q-PCR验证显示与正常组比,GDM�Objective To investigate the molecular mechanism of abnormal nervous system development in gestational diabetes mellitus(GDM)in mice and to verify the expression of miRNA-7047-5p.Methods From May 2020 to December 2021,a total of 30 KM mice(20 females and 10 males)were caged overnight in a ratio of male to female 1:2 to construct pregnant mice.Pregnant mice were randomly divided into model group and control group,and the GDM mice model was established by intraperitoneal injection of streptozotocin(STZ).Random blood glucose was detected by glucose meter.The whole structure and morphology of brain tissue were observed under anatomical microscope.The morphology of hippocampal cells was observed by hematoxylin-eosin(HE)staining.The differentially expressed miRNA genes in the mice hippocampus were detected by microarray assay.Differentially expressed miRNA genes were analyzed by Kyoto Encyclopedia of Genes and Genomes(KEGG).The expression of miRNA-7047-5p was detected by quantitative polymerase chain reaction(q-PCR).Results Compared with the control group,the blood sugar of the model group mice increased on the next day after administration(E1.5).The blood glucose values of E1.5,E5.5,E11.5,and E18.5 were(5.67±1.06),(6.84±1.23),(5.61±1.04),(12.08±2.16),(5.49±1.05),(14.27±2.71),(5.53±1.03),(15.14±2.86)mmol/L(P<0.05);Starting from the first day after administration,the random blood glucose levels in the model group mice increased.On the first day,second day,and third day,E4.5,E10.5,and E17.5 after administration,the random blood glucose levels were(5.83±1.06),(22.36±3.74),(5.79±1.03),(20.65±3.49),(5.95±1.11),(19.94±2.97),(5.93±1.14),(18.05±3.14),(5.87±1.15),(16.49±2.85),(5.69±1.03),(14.89±2.37)mmol/L(P<0.05).In the control group,the whole brain tissue was not swollen,and the sulci and gyri were clear.The sulci and gyri in GDM group were shallow.In the control group,the cells in the CA1 area of the hippocampus were closely arranged and orderly,full in shape,with clear nucleoli and no nucleolar pyknosis.

关 键 词：糖尿病 妊娠 基因芯片技术 子代小鼠 神经系统发育异常 微小核糖核酸-7047-5p 

分 类 号：R714.256[医药卫生—妇产科学]