Objective Placental dysfunction induced by fetal cardiopulmonary bypass(CPB)imposes limitations on the clinical application of this procedure.The potential impact of microRNA-mediated autophagy in placental endothelia...Objective Placental dysfunction induced by fetal cardiopulmonary bypass(CPB)imposes limitations on the clinical application of this procedure.The potential impact of microRNA-mediated autophagy in placental endothelial cells on overall placental function remains elusive,necessitating a comprehensive exploration of the underlying mechanisms involved.Methods We established fetal sheep CPB models and employed immunohistochemistry to assess the placental expression of ATG7.Bioinformatic analysis,coupled with dual-luciferase reporter assays,was used to elucidate the intricate relationship between miR-320a and ATG7.Changes in ATG7 expression were further investigated through Western blotting and quantitative polymerase chain reaction(qPCR).Human umbilical vein endothelial cells(HUVECs)were cultured,and in vitro experiments were conducted to evaluate their regulatory effects on endothelial function.Immunoblotting was used to measure the expression levels of ATG7,endothelin-1(ET-1),SIRT1,and FOXO1,whereas enzyme-linked immunosorbent assay(ELISA)was used to quantify nitric oxide(NO)production.Results Sixty minutes after CPB,a substantial decrease in ATG7 expression in placental tissue was observed.The downregulation of ATG7 expression led to increased ET-1 production in HUVECs,concomitant with decreased NO production.miR-320a was identified as a specific regulator of ATG7 expression,with subsequent experiments demonstrating a significant reduction in placental ATG7 levels upon injection of the miR-320a agomir compared with the miR-320a antagomir during fetal sheep CPB.In HUVECs,miR-320a downregulated ATG7,resulting in increased ET-1 production and diminished NO production.Treatment with the miR-320a mimic/miR-320a inhibitor revealed that miR-320a inhibited the SIRT1/FOXO1 pathway in HUVECs by downregulating ATG7 expression,culminating in increased ET-1 production and reduced NO levels.Conclusion The observed downregulation of placental ATG7 expression subsequent to fetal CPB is intricately associated with endothelial dysfunction.Furthermore,our findings underscore the specific regulatory role of miR-320a in modulating ATG7 expression within the placenta.At the cellular level,increasing the level of miR-320a has emerged as a potential strategy for modulating endothelial function through the inhibition of ATG7 and the SIRT1/FOXO1 pathway.展开更多
目的:探讨糖尿病肾病(DKD)患者血清Beclin1、ATG7水平与蛋白尿的相关性。方法:收集2018年12月—2019年10月在本院门诊或住院部诊断为DKD共108例患者的临床资料,按ACR分为正常白蛋白尿组、微量白蛋白尿组和大量白蛋白尿组。纳入同期我院...目的:探讨糖尿病肾病(DKD)患者血清Beclin1、ATG7水平与蛋白尿的相关性。方法:收集2018年12月—2019年10月在本院门诊或住院部诊断为DKD共108例患者的临床资料,按ACR分为正常白蛋白尿组、微量白蛋白尿组和大量白蛋白尿组。纳入同期我院体检健康者共30例为对照组。检测血清Beclin1、ATG7水平,用Pearson/Spearman相关分析Beclin1、ATG7与尿微量白蛋白、尿微量白蛋白/肌酐等指标的相关性。用多重线性逐步回归法分析Beclin1、ATG7的影响因素。结果:(1)与对照组相比,DKD三组患者血清Beclin1、ATG7水平均降低,其中大量白蛋白尿组最低(P<0.001)。(2)Beclin1与HDL-C、eGFR、Hb、Alb呈正相关(P<0.05),与糖尿病病程、HbAlc、PBG、TC、BUN、Scr、UA、UAlb、ACR、24 h UTP呈负相关(P<0.05)。ATG7与HDL-C、eGFR、Hb、Alb呈正相关(P<0.05),与糖尿病病程、FBG、HbAlc、PBG、TC、LDL-C、BUN、Scr、UA、UAlb、ACR、24 h UTP呈负相关(P<0.05)。(3)经多重线性逐步回归分析,Beclin1水平的重要预测指标为糖尿病病程、ACR、eGFR、HDL-C(P<0.01)。ATG7水平的预测指标为ACR、eGFR、HbAlc(P<0.05)。结论:本研究发现DKD患者血清中自噬相关蛋白Beclin1、ATG7水平降低,与蛋白尿程度存在相关性,对于明确自噬障碍与DKD进展可能具有一定的研究价值,其具体机制有待于进一步研究。展开更多
目的探讨MNX1通过调控ATG7诱导细胞自噬介导鼻咽癌放疗抵抗的作用机制。方法采用人鼻咽癌抗辐射细胞系HONE-1-IR和辐射敏感细胞系HONE-1进行体外分析;检测细胞中MNX1的表达水平,比较自噬标志物LC3Ⅱ、ATG7和p62表达情况;并分析MNX1与ATG...目的探讨MNX1通过调控ATG7诱导细胞自噬介导鼻咽癌放疗抵抗的作用机制。方法采用人鼻咽癌抗辐射细胞系HONE-1-IR和辐射敏感细胞系HONE-1进行体外分析;检测细胞中MNX1的表达水平,比较自噬标志物LC3Ⅱ、ATG7和p62表达情况;并分析MNX1与ATG7调控细胞自噬的关系。结果与HONE-1细胞相比,HONE-1-IR细胞中MNX1 m RNA和蛋白水平均升高,LC3Ⅱ、ATG7和p62蛋白表达水平同样升高(P<0.05);下调MNX1抑制了自噬体的形成(P<0.05)。与阴性对照组相比,转染si MNX1的细胞增殖率和细胞活力明显降低(P<0.05);细胞集落形成率明显降低(P<0.05)。下调MNX1明显阻断了自噬体的形成,上调ATG7后,自噬体的抑制水平恢复到对照组的水平(P<0.05);在HONE-1-IR细胞中通过下调MNX1抑制了细胞的增殖和活力,而通过上调ATG7逆转了这种抑制(P<0.05)。结论MNX1通过上调ATG7调控细胞自噬参与放疗抵抗,通过敲低MNX1-ATG7轴可提高放疗抵抗细胞的敏感性,MNX1可作为增强放射敏感性的潜在治疗靶点。展开更多
Background:Myocardial infarction(MI)is an acute condition in which the heart mus-cle dies due to the lack of blood supply.Previous research has suggested that au-tophagy and angiogenesis play vital roles in the preven...Background:Myocardial infarction(MI)is an acute condition in which the heart mus-cle dies due to the lack of blood supply.Previous research has suggested that au-tophagy and angiogenesis play vital roles in the prevention of heart failure after MI,and miR-106a is considered to be an important regulatory factor in MI.But the specific mechanism remains unknown.In this study,using cultured venous endothelial cells and a rat model of MI,we aimed to identify the potential target genes of miR-106a and discover the mechanisms of inhibiting autophagy and angiogenesis.Methods:We first explored the biological functions of miR-106a on autophagy and angiogenesis on endothelial cells.Then we identified ATG7,which was the down-stream target gene of miR-106a.The expression of miR-106a and ATG7 was investi-gated in the rat model of MI.Results:We found that miR-106a inhibits the proliferation,cell cycle,autophagy and angiogenesis,but promoted the apoptosis of vein endothelial cells.Moreover,ATG7 was identified as the target of miR-106a,and ATG7 rescued the inhibition of autophagy and angiogenesis by miR-106a.The expression of miR-106a in the rat model of MI was decreased but the expression of ATG7 was increased in the infarction areas.Conclusion:Our results indicate that miR-106a may inhibit autophagy and angiogenesis by targeting ATG7.This mechanism may be a potential therapeutic treatment for MI.展开更多
基金supported by the National Key Research and Development Program of China(No.2022YFC2407406)Guangzhou Science and Technology Planning Project(No.2023B03J0596)2023 Stability Support for Innovative Capacity Building of Guangdong Provincial Scientific Research Institutions(No.KD022023019).
文摘Objective Placental dysfunction induced by fetal cardiopulmonary bypass(CPB)imposes limitations on the clinical application of this procedure.The potential impact of microRNA-mediated autophagy in placental endothelial cells on overall placental function remains elusive,necessitating a comprehensive exploration of the underlying mechanisms involved.Methods We established fetal sheep CPB models and employed immunohistochemistry to assess the placental expression of ATG7.Bioinformatic analysis,coupled with dual-luciferase reporter assays,was used to elucidate the intricate relationship between miR-320a and ATG7.Changes in ATG7 expression were further investigated through Western blotting and quantitative polymerase chain reaction(qPCR).Human umbilical vein endothelial cells(HUVECs)were cultured,and in vitro experiments were conducted to evaluate their regulatory effects on endothelial function.Immunoblotting was used to measure the expression levels of ATG7,endothelin-1(ET-1),SIRT1,and FOXO1,whereas enzyme-linked immunosorbent assay(ELISA)was used to quantify nitric oxide(NO)production.Results Sixty minutes after CPB,a substantial decrease in ATG7 expression in placental tissue was observed.The downregulation of ATG7 expression led to increased ET-1 production in HUVECs,concomitant with decreased NO production.miR-320a was identified as a specific regulator of ATG7 expression,with subsequent experiments demonstrating a significant reduction in placental ATG7 levels upon injection of the miR-320a agomir compared with the miR-320a antagomir during fetal sheep CPB.In HUVECs,miR-320a downregulated ATG7,resulting in increased ET-1 production and diminished NO production.Treatment with the miR-320a mimic/miR-320a inhibitor revealed that miR-320a inhibited the SIRT1/FOXO1 pathway in HUVECs by downregulating ATG7 expression,culminating in increased ET-1 production and reduced NO levels.Conclusion The observed downregulation of placental ATG7 expression subsequent to fetal CPB is intricately associated with endothelial dysfunction.Furthermore,our findings underscore the specific regulatory role of miR-320a in modulating ATG7 expression within the placenta.At the cellular level,increasing the level of miR-320a has emerged as a potential strategy for modulating endothelial function through the inhibition of ATG7 and the SIRT1/FOXO1 pathway.
文摘目的:探讨糖尿病肾病(DKD)患者血清Beclin1、ATG7水平与蛋白尿的相关性。方法:收集2018年12月—2019年10月在本院门诊或住院部诊断为DKD共108例患者的临床资料,按ACR分为正常白蛋白尿组、微量白蛋白尿组和大量白蛋白尿组。纳入同期我院体检健康者共30例为对照组。检测血清Beclin1、ATG7水平,用Pearson/Spearman相关分析Beclin1、ATG7与尿微量白蛋白、尿微量白蛋白/肌酐等指标的相关性。用多重线性逐步回归法分析Beclin1、ATG7的影响因素。结果:(1)与对照组相比,DKD三组患者血清Beclin1、ATG7水平均降低,其中大量白蛋白尿组最低(P<0.001)。(2)Beclin1与HDL-C、eGFR、Hb、Alb呈正相关(P<0.05),与糖尿病病程、HbAlc、PBG、TC、BUN、Scr、UA、UAlb、ACR、24 h UTP呈负相关(P<0.05)。ATG7与HDL-C、eGFR、Hb、Alb呈正相关(P<0.05),与糖尿病病程、FBG、HbAlc、PBG、TC、LDL-C、BUN、Scr、UA、UAlb、ACR、24 h UTP呈负相关(P<0.05)。(3)经多重线性逐步回归分析,Beclin1水平的重要预测指标为糖尿病病程、ACR、eGFR、HDL-C(P<0.01)。ATG7水平的预测指标为ACR、eGFR、HbAlc(P<0.05)。结论:本研究发现DKD患者血清中自噬相关蛋白Beclin1、ATG7水平降低,与蛋白尿程度存在相关性,对于明确自噬障碍与DKD进展可能具有一定的研究价值,其具体机制有待于进一步研究。
文摘目的探讨MNX1通过调控ATG7诱导细胞自噬介导鼻咽癌放疗抵抗的作用机制。方法采用人鼻咽癌抗辐射细胞系HONE-1-IR和辐射敏感细胞系HONE-1进行体外分析;检测细胞中MNX1的表达水平,比较自噬标志物LC3Ⅱ、ATG7和p62表达情况;并分析MNX1与ATG7调控细胞自噬的关系。结果与HONE-1细胞相比,HONE-1-IR细胞中MNX1 m RNA和蛋白水平均升高,LC3Ⅱ、ATG7和p62蛋白表达水平同样升高(P<0.05);下调MNX1抑制了自噬体的形成(P<0.05)。与阴性对照组相比,转染si MNX1的细胞增殖率和细胞活力明显降低(P<0.05);细胞集落形成率明显降低(P<0.05)。下调MNX1明显阻断了自噬体的形成,上调ATG7后,自噬体的抑制水平恢复到对照组的水平(P<0.05);在HONE-1-IR细胞中通过下调MNX1抑制了细胞的增殖和活力,而通过上调ATG7逆转了这种抑制(P<0.05)。结论MNX1通过上调ATG7调控细胞自噬参与放疗抵抗,通过敲低MNX1-ATG7轴可提高放疗抵抗细胞的敏感性,MNX1可作为增强放射敏感性的潜在治疗靶点。
基金National Natural Science Foundation of China,Grant/Award Number:32070542Guangdong Basic and Applied Basic Research Foundation,Grant/Award Number:2021A1515010873 and 2022A1515011455+1 种基金Breed Industry Innovation Park of Guangdong Xiaoerhua Pig,Grant/Award Number:2022-4408X1-43010402-0019Hainan Provincial Natural Science Foundation,Grant/Award Number:818MS132。
文摘Background:Myocardial infarction(MI)is an acute condition in which the heart mus-cle dies due to the lack of blood supply.Previous research has suggested that au-tophagy and angiogenesis play vital roles in the prevention of heart failure after MI,and miR-106a is considered to be an important regulatory factor in MI.But the specific mechanism remains unknown.In this study,using cultured venous endothelial cells and a rat model of MI,we aimed to identify the potential target genes of miR-106a and discover the mechanisms of inhibiting autophagy and angiogenesis.Methods:We first explored the biological functions of miR-106a on autophagy and angiogenesis on endothelial cells.Then we identified ATG7,which was the down-stream target gene of miR-106a.The expression of miR-106a and ATG7 was investi-gated in the rat model of MI.Results:We found that miR-106a inhibits the proliferation,cell cycle,autophagy and angiogenesis,but promoted the apoptosis of vein endothelial cells.Moreover,ATG7 was identified as the target of miR-106a,and ATG7 rescued the inhibition of autophagy and angiogenesis by miR-106a.The expression of miR-106a in the rat model of MI was decreased but the expression of ATG7 was increased in the infarction areas.Conclusion:Our results indicate that miR-106a may inhibit autophagy and angiogenesis by targeting ATG7.This mechanism may be a potential therapeutic treatment for MI.
