Background Higher embryonic mortality,especially in aged breeding hens,is associated with insufficient hepatic functionality in maintaining redox homeostasis.Our previous study demonstrated that egg exosome-derived mi...Background Higher embryonic mortality,especially in aged breeding hens,is associated with insufficient hepatic functionality in maintaining redox homeostasis.Our previous study demonstrated that egg exosome-derived miRNAs may play a key role in modulating embryonic oxidation-reduction process,whereas the exact function and mechanism were still poorly understood.The present study aimed to investigate the roles of egg exosome miRNAs in maintaining dynamic equilibrium of free radicals and peroxide agents in embryonic liver,as well as demonstrate the specific mechanism using oxidative stress-challenged hepatocytes.Results Compared to 36-week-old breeding hens,decreased hatchability and increased embryonic mortality were observed in 65-week-old breeding hens.Meanwhile,the older group showed the increased MDA levels and decreased SOD and GSH-Px activities in embryonic liver,muscle and serum.Embryonic mortality was significantly positively correlated with MDA level and negatively correlated with GSH-Px activity in embryonic liver.In addition,363 differentially expressed genes(DEGs)were identified in embryonic liver,13 differentially expressed miRNAs(DE-miRNAs)were identified in egg exosomes.These DEGs and DE-miRNAs were involved in oxidoreductase activity,glutathione metabolic process,MAPK signaling pathway,apoptosis and autophagy.miRNA-mRNA network analysis further found that DEGs targeted by DE-miRNAs were mainly enriched in programmed cell death,such as apoptosis and autophagy.Wherein,MAPK10 with highest MCC and AUC values was significantly related to GSH-Px activity and MDA level,and served as the target gene of miR-145-5p based on dual luciferase reporter experiment and correlation analysis.Bioinformatics analysis found that miR-145-5p/MAPK10 axis might alleviate peroxide generation and apoptosis.In primary hepatocytes of chick embryos,miR-145-5p transfection significantly reversed H_(2)O_(2)-induced mitochondrial ROS increase,MAPK10,BAX and CASP3 overexpression and excessive apoptosis.Conclusion Exosome miR-145-5p in eggs could target MAPK10 and decrease mitochondrial ROS,attenuating oxidative damage and apoptosis in hepatocytes of chick embryos.These findings may provide new theoretical basis for the improvement of maternal physiological status to maintain embryonic redox homeostasis by nutritional or genetic modifications.展开更多
基金supported by China Agriculture Research System of MOF and MARA(CARS-40)the National Natural Science Foundation of China(32302776)。
文摘Background Higher embryonic mortality,especially in aged breeding hens,is associated with insufficient hepatic functionality in maintaining redox homeostasis.Our previous study demonstrated that egg exosome-derived miRNAs may play a key role in modulating embryonic oxidation-reduction process,whereas the exact function and mechanism were still poorly understood.The present study aimed to investigate the roles of egg exosome miRNAs in maintaining dynamic equilibrium of free radicals and peroxide agents in embryonic liver,as well as demonstrate the specific mechanism using oxidative stress-challenged hepatocytes.Results Compared to 36-week-old breeding hens,decreased hatchability and increased embryonic mortality were observed in 65-week-old breeding hens.Meanwhile,the older group showed the increased MDA levels and decreased SOD and GSH-Px activities in embryonic liver,muscle and serum.Embryonic mortality was significantly positively correlated with MDA level and negatively correlated with GSH-Px activity in embryonic liver.In addition,363 differentially expressed genes(DEGs)were identified in embryonic liver,13 differentially expressed miRNAs(DE-miRNAs)were identified in egg exosomes.These DEGs and DE-miRNAs were involved in oxidoreductase activity,glutathione metabolic process,MAPK signaling pathway,apoptosis and autophagy.miRNA-mRNA network analysis further found that DEGs targeted by DE-miRNAs were mainly enriched in programmed cell death,such as apoptosis and autophagy.Wherein,MAPK10 with highest MCC and AUC values was significantly related to GSH-Px activity and MDA level,and served as the target gene of miR-145-5p based on dual luciferase reporter experiment and correlation analysis.Bioinformatics analysis found that miR-145-5p/MAPK10 axis might alleviate peroxide generation and apoptosis.In primary hepatocytes of chick embryos,miR-145-5p transfection significantly reversed H_(2)O_(2)-induced mitochondrial ROS increase,MAPK10,BAX and CASP3 overexpression and excessive apoptosis.Conclusion Exosome miR-145-5p in eggs could target MAPK10 and decrease mitochondrial ROS,attenuating oxidative damage and apoptosis in hepatocytes of chick embryos.These findings may provide new theoretical basis for the improvement of maternal physiological status to maintain embryonic redox homeostasis by nutritional or genetic modifications.
文摘目的探究急性ST段抬高型心肌梗死(ST-segment elevation myocardial infarction,STEMI)患者外周血微小核糖核酸(microRNA,miRNA)-145-5p、miR-27b-3p浓度对经皮冠状动脉介入(percutaneous coronary intervention,PCI)治疗后并发心力衰竭(heart failure,HF)的预测价值。方法选择于2021年1月至2023年2月于成都市第五人民医院就诊的STEMI患者176例(均行PCI治疗)作为观察对象,观察STEMI患者PCI治疗后HF的发生情况,分为发生组和未发生组。实时定量聚合酶链反应(quantitative real time polymerase chain reaction,qRT-PCR)法检测外周血miR-145-5p、miR-27b-3p浓度。Pearson法、Spearman法分析血清miR-145-5p、miR-27b-3p及二者与一般资料的相关性。STEMI患者发生HF的影响因素采用多因素Logistic回归分析。绘制受试者工作特征曲线(receiver operating characteristic curve,ROC)分析血清miR-145-5p、miR-27b-3p对STEMI患者PCI治疗后发生HF的预测价值。结果两组患者的病变支数、血清同型半胱氨酸(homocysteine,Hcy)、超敏C反应蛋白(hypersensitive C-reactive protein,hs-CRP)浓度比较,差异有统计学意义(P<0.05)。发生组患者的血清miR-145-5p、miR-27b-3p浓度高于未发生组,差异有统计学意义(P<0.05)。根据Spearman相关性分析得知,血清miR-145-5p、miR-27b-3p浓度与病变支数呈正相关(P<0.05)。根据Pearson相关性分析得知,血清miR-145-5p浓度与miR-27b-3p浓度呈正相关(P<0.05);血清miR-145-5p、miR-27b-3p浓度与Hcy、hs-CRP浓度呈正相关(P<0.05)。多因素Logistic回归分析显示多支病变、Hcy、hs-CRP、miR-145-5p、miR-27b-3p为影响STEMI患者PCI治疗后发生HF的危险因素(P<0.05)。血清miR-145-5p预测STEMI患者PCI治疗后发生HF的曲线下面积(area under the curve,AUC)为0.896,血清miR-27b-3p预测STEMI患者PCI治疗后发生HF的AUC为0.883,二者联合预测STEMI患者发生HF的AUC为0.962,二者联合优于各自单独预测(P<0.05)。结论miR-145-5p、miR-27b-3p浓度升高与STEMI患者PCI治疗后发生HF相关,两者联合对STEMI患者PCI治疗后发生HF具有较高的预测价值。
