Objective To study the effects of gestational diabetes (GDM) on morphological structure of brain tissue and microribonucleotide (miRNA) expression profile in neonatal mice, and to provide a new research target for the prevention and treatment of abnormal neurodevelopment in GDM progeny. Methods The pregnant mice were divided into model group and control group, each group consisted of 10 mice. The model group mice established a GDM model by injecting streptozotocin to measure fasting blood glucose (FPG) and random blood glucose (GLU) at different times. Successful molded mice were randomly divided into model group A and model group C, and control mice were divided into control group B and control group D, with 5 mice in each group.The newborn mice in groups A and B were used for hippocampal tissue GeneChip detection and brain morphology structure observation, and group C and D newborn mice were used for qRT-PCR detection of hippocampus tissue expression differences to verify the differentially expressed genes of miRANs obtained by GeneChip screening.After giving birth, the neonatal mice were sacrificed by decapitation, and the brain tissue was dissected to observe the overall morphological structure. The structural changes of hippocampus were observed under HE chromogenic microscope. The Agilent mouse miRNA oligonucleotide gene chip was used to detect the miRNA expression profile of mouse hippocampus, screen differential miRNAs and predict their target genes, and conduct GO analysis and signal transduction pathway analysis of target genes. The relative expression levels of the screened miRNAs were verified by qRT-PCR. Results Compared with the control group, the GLU increased significantly from the 3rd day after drug administration in the model group (P<0.01). Macroscopic observation of control group B mice had normal brain morphology and structure, smooth appearance, clear gyrus, close arrangement of hippocampus cell structure, uniform staining and complete structure; in model group A, the number of hippocampus cells decreased, loose arrangement and deep staining. In the initial screen of miRNA microarray, there were 11 differentially expressed miRNAs between control and model groups, all of which were downregulated miRNAs, including let-7b-5p、miR-130b-3p、miR-181c-5p、miR-181d-5p、miR-3099-3p、miR-3470a、miR-3473a、miR-3473b、miR-500-3p、miR-532-5p、miR-7047-5p(P＜0.05)。Two miRNAs (miR-3473b, miR-7047-75p) and 5 target genes (MAPK3, MAPK11, MAPK14, CALM3, AKT3). The relative expression of miR-3473b and miR-7047-5p in model group C were lower than that in control group D (t=19.13 and 6.24, P<0.05), and the validation results were consistent with the microarray test results. Conclusion Compared with the offspring of normal pregnant mice, GDM offspring mice have abnormal development of brain structure and damage of hippocampal nerve cells, and there are a large number of abnormal expression of miRNAs in hippocampal tissue. Differentially expressed miRNAs can be used as research targets for prevention and treatment of GDM offspring neurodevelopmental abnormalities.
Key words
gestational diabetes /
microRNA /
morphological structure of brain tissue /
gene expression profile
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References
[1] 郑晓芳, 彭娘海, 黄真轩,等. BDNF在妊娠糖尿病小鼠胎盘中的表达及对小鼠子代宫内生长发育情况的影响[J].中国优生与遗传杂志, 2022, 30(2):196-201.
Zheng XF, Peng NH, Huang ZX, et al.The expression of BDNF in the placenta of pregnant diabetes mice and its effect on the intrauterine growth and development of the offspring of mice[J].Chin J Birth Health Heredity, 2022,30(2):196-201.
[2] Cao T, Zhen XC.Dysregulation of miRNA and its potential therapeutic application in schizophrenia[J]. CNS Neurosci Ther, 2018, 24(7):586-597.
[3] 周丽媛, 肖新华. 生命早期营养与糖代谢异常发生发展的新启示:糖尿病防治窗口需前移[J].中华糖尿病杂志, 2021, 13(12):1109-1114.
Zhou LY, Xiao XH. New enlightenment from the occurrence and development of nutrition and glucose metabolism abnormalities in early life:The window for prevention and treatment of diabetes needs to move forward[J].Chin J Diabetes Mellitus, 2021,13 (12): 1109-1114.
[4] 何泽银,邓皓元,李汇华.miRNA-27 a在心血管疾病中的作用机制[J].生理科学进展,2021,52(1):44-48.
He ZY, Deng HY, Li HH. The mechanism of action of miRNA-27a in cardiovascular disease[J]. Prog Physiol Sci, 2021,52 (1): 44-48.
[5] 红梅,张凤兰,戴海龙.MiRNAs在成体神经发生中的作用[J].临床与病理杂志,2021,41(3):663-672.
Hong M, Zhang FL, Dai HL. The role ofMiRNAs in adult neurogenesis[J]. J Clin Pathol Res, 2021,41 (3): 663-672.(in Chinese)
[6] Mathoux J, Henshall DC, Brennan GP. Regulatory mechanisms of the RNA modification m6A and significance in brain function in health and disease[J]. Front Cell Neurosci, 2021, 15:671932.
[7] 丁敬,冷在华,张秀琼.人参皂苷Re通过内质网应激通路对妊娠期糖尿病大鼠妊娠结局的影响及相关机制[J].临床和实验医学杂志,2021, 20(8):804-808.
Ding J,Leng ZH, Zhang XQ. The effect of ginsenoside Re on the pregnancy outcome of pregnant diabetes rats through endoplasmic reticulum stress pathway and related mechanisms[J]. J Lab Clin Med, 2021, 20 (8): 804-808.(in Chinese)
[8] 刘侃,王秋明,宋婉玉,等. 口服益生菌对妊娠期糖尿病小鼠肠道菌群及调节性免疫细胞的影响[J]. 中国微生态学杂志, 2020, 32(2):166-171.
Liu K, Wang QM, Song WY, et al. Effects of oral probiotics on intestinal flora and regulatory immune cells in pregnant diabetes mice[J]Chin J Microecol, 2020, 32 (2): 166-171.(in Chinese)
[9] Nguyen CL, Pham NM,Binns CW,et al. Prevalence of gestational diabetes mellitus in eastern and southeastern Asia: A systematic review and meta-analysis[J]. J Diabetes Res,2018, 2018:6536974.
[10] American Diabetes Association 2. Classification and diagnosis of diabetes: Standards of medical care in diabetes—2020[J]. Diabetes Care, 2020, 43(Suppl):14-31.
[11] Vounzoulaki E, Khunti K, Abner SC, et al. Progression to type 2 diabetes in women with a known history of gestational diabetes: Systematic review and Meta-analysis[J]. BMJ, 2020, 369:m1361.
[12] Li C, Zhou P,Cai Y, et al Associations between gestational diabetes mellitus and the neurodevelopment of offspring from 1 month to 72 months: Study protocol for a cohort study[J]. BMJ Open, 2020,10(11):e040305
[13] Veena SR, Krishnaveni GV, Srinivasan K, et al. Childhood cognitive ability: Relationship to gestational diabetes mellitus in India[J]. Diabetologia, 2010, 53(10):2134-2138.
[14] Lester BM,Marsit CJ. Epigenetic mechanisms in the placenta related to infant neurodevelopment[J]. Epigenomics, 2018, 10(3):321-333.
[15] Channakkar AS, Singh T, Pattnaik B, et al. MiRNA-137-mediated modulation of mitochondrial dynamics regulates human neural stem cell fate[J]. Stem Cells, 2020, 38(5):683-697.
[16] Wang X, Chen S, Ni J, et al. miRNA-3473b contributes to neuroinflammation following cerebral ischemia[J]. Cell Death & Disease, 2018, 9(1): 1-13.
[17] Yang X,Xie J, Jia L, et al. Analysis of miRNAs involved in mouse brain damage upon enterovirus 71 infection[J]. Front Cell Infect Mi, 2017, 7: 133.
[18] Cao S, Yang Y, Yu Q, et al. Electroacupuncture alleviates ischaemic brain injury by regulating the miRNA-34/Wnt/autophagy axis[J].Brb Medline, 2021, 170: 155-161.
[19] Lv Q, Zhong Z, Hu B, et al. MicroRNA-3473b regulates the expression of TREM2/ULK1 and inhibits autophagy in inflammatory pathogenesis of Parkinson disease[J].J Neurochem, 2021, 157(3): 599-610.
[20] Wu C,Xue Y, Wang P, et al. IFN-γ primes macrophage activation by increasing phosphatase and tensin homolog via downregulation of miR-3473b[J].J Immunol, 2014,193(6):3036-3044.
[21] Andrea L,Silvia AC,Murdocca M, et al. A perturbed microRNA expression pattern characterizes embryonic neural stem cells derived from a severe mouse model of spinal muscular atrophy (SMA)[J] .Int J Mol Sci, 2015, 16: 18312-18327.
[22] Park So-Yeon,Kim Da-Seul,Kim Hyun-Mun,et al. Human mesenchymal stem cell-derived extracellular vesicles promote neural differentiation of neural progenitor cells[J].Int J Mol Sci,2022,23,7047.
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