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作 者:宋琦 杜志欣 杨丽萍[1] 侯俊林[1] 赵佳佳 吴永叶 王耀辉 李笑琳 SONG Qi;DU Zhixin;YANG Liping;HOU Junlin;ZHAO Jiajia;WU Yongye;WANG Yaohui;LI Xiaolin(Henan University of Chinese Medicine School of Traditional Chinese Medicine,Zhengzhou 450046,China)
出 处:《中国实验动物学报》2024年第7期828-835,共8页Acta Laboratorium Animalis Scientia Sinica
基 金:国家自然科学基金(81973596)。
摘 要:目的 探讨孕期恐惧应激对胎盘葡萄糖营养物转运体(glucose transporter, GLUT)表达的影响,为孕期应激致胎损理论提供证据。方法 将20只Wistar孕鼠,随机分为对照组和模型组,每组10只。其中,模型组采取旁观电击法造模持续19 d,对照组正常饲养。实验结束后记录胎产数量、胎盘与胎鼠重量,计算胎盘效率。透视电镜观察胎盘细胞形态学变化。生物信息学分析筛选孕期恐惧应激胎盘的差异基因,进行基因富集分析(gene set enrichment analysis, GSEA)。蛋白质免疫印迹法(Western Blot)、实时荧光定量聚合酶链式反应(Real-time PCR)、免疫组化法检测葡萄糖转运蛋白1(glucosetransporter 1,GLUT1)、葡萄糖转运蛋白3(GLUT3)、葡萄糖转运蛋白6(GLUT6)和葡萄糖转运蛋白7(GLUT7)表达水平。结果 与对照组相比,模型组胎产数量和胎盘效率显著降低;模型组胎盘微绒毛稀疏短少,线粒体及内质网肿胀;GSEA分析发现,模型组胎盘基因显著富集在细胞葡萄糖稳态;Western Blot、Real-time PCR和免疫组化结果表明,模型组胎盘中GLUT1、GLUT6以及GLUT7表达均下降。结论 孕期恐惧应激暴露导致不良胎产结局,可能与胎盘中GLUT1、GLUT6、GLUT7三种关键葡萄糖转运体降低有关。Objective To investigate the effect of fear-induced stress during pregnancy on the expression of glucose transporters(GLUT)in the placenta,providing evidence for the theory of fetal damage caused by fear-induced stress during pregnancy.Methods Twenty pregnant Wistar rats were randomly divided into a control group and a model group of 10 rats each.In the model group,a fear-induced stress model was established using the modified bystander electroshock method for 20 days.After the experiment,the number of offspring and the weights of the placenta and fetal rats were measured,and the placental efficiency was calculated.Transmission electron microscopy was used to observe the morphological changes of placental cells.Bioinformatics analysis was performed to screen for differential genes in placentas affected by pregnancy stress-phobia,and gene set enrichment analysis was performed.Protein immunoblotting(Western Blot),Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR),and immunohistochemistry were used to detect the expression levels of GLUT1,GLUT3,GLUT6,and GLUT7 proteins and genes.Results The placental efficiency was significantly reduced in the model group compared with that in the control group.The result of transmission electron microscopy in the model group showed that the placental microvilli were sparse and short and that the mitochondria and endoplasmic reticulum were swollen.Gene set enrichment analysis revealed that placental genes were significantly enriched in cellular glucose homeostasis in the model group compared with those in the control group.The result of Western Blot,Real-time PCR,and immunohistochemistry indicated a decrease in both the protein and gene expression levels of GLUT1,GLUT6,and GLUT7 in the placenta of pregnant rats.Conclusions Prenatal exposure to fear-induced stress may lead to adverse pregnancy outcomes.These adverse outcomes are potentially associated with reduced levels of three key GLUTs in the placenta:GLUT1,GLUT6,and GLUT7.
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