BACKGROUND Diabetic kidney disease(DKD),characterized by increased urinary microalbumin levels and decreased renal function,is the primary cause of end-stage renal di-sease.Its pathological mechanisms are complicated ...BACKGROUND Diabetic kidney disease(DKD),characterized by increased urinary microalbumin levels and decreased renal function,is the primary cause of end-stage renal di-sease.Its pathological mechanisms are complicated and multifactorial;Therefore,sensitive and specific biomarkers are needed.Urinary exosome originate from diverse renal cells in nephron segments and partially mirror the pathological changes in the kidney.The microRNAs(miRNAs)in urinary exosome are remark-ably stable and highly tissue-specific for the kidney.METHODS Type 2 diabetic mellitus(T2DM)patients were recruited from the Second Hospital of Hebei Medical University and were divided into two groups:DM,diabetic pa-tients without albuminuria[urinary albumin to creatinine ratio(UACR)<30 mg/g]and DKD,diabetic patients with albuminuria(UACR≥30 mg/g).Healthy subjects were the normal control(NC)group.Urinary exosomal miR-145-5p,miR-27a-3p,and miR-29c-3p,were detected using real-time quantitative polymerase chain reaction.The correlation between exosomal miRNAs and the clinical in-dexes was evaluated.The diagnostic values of exosomal miR-145-5p and miR-27a-3p in DKD were determined using receiver operating characteristic(ROC)analysis.Biological functions of miR-145-5p were investigated by performing RESULTS Urinary exosomal expression of miR-145-5p and miR-27a-3p was more upregulated in the DKD group than in the DM group(miR-145-5p:4.54±1.45 vs 1.95±0.93,P<0.001;miR-27a-3p:2.33±0.79 vs 1.71±0.76,P<0.05)and the NC group(miR-145-5p:4.54±1.45 vs 1.55±0.83,P<0.001;miR-27a-3p:2.33±0.79 vs 1.10±0.51,P<0.001).The exosomal miR-145-5p and miR-27a-3p positively correlated with albuminuria and serum creatinine and negatively correlated with the estimated glomerular filtration rate.miR-27a-3p was also closely related to blood glucose,gly-cosylated hemoglobin A1c,and low-density lipoprotein cholesterol.ROC analysis revealed that miR-145-5p had a better area under the curve of 0.88[95%confidence interval(CI):0.784-0.985,P<0.0001]in diagnosing DKD than miR-27a-3p with 0.71(95%CI:0.547-0.871,P=0.0239).Bioinformatics analysis revealed that the target genes of miR-145-5p were located in the actin filament,cytoskeleton,and extracellular exosome and were involved in the pathological processes of DKD,including apoptosis,inflammation,and fibrosis.CONCLUSION Urinary exosomal miR-145-5p and miR-27a-3p may serve as novel noninvasive diagnostic biomarkers or promising therapeutic targets for DKD.展开更多
目的研究miRNA-21-5p对光诱导的人视网膜色素上皮细胞氧化应激损伤的影响。方法将体外培养的人视网膜色素上皮细胞系ARPE-19细胞随机分为对照组(TransIntro EL Transfection Reagent转染液培养)、损伤组(TransIntro EL Transfection Rea...目的研究miRNA-21-5p对光诱导的人视网膜色素上皮细胞氧化应激损伤的影响。方法将体外培养的人视网膜色素上皮细胞系ARPE-19细胞随机分为对照组(TransIntro EL Transfection Reagent转染液培养)、损伤组(TransIntro EL Transfection Reagent转染液+光损伤)、过表达组(TransIntro EL Transfection Reagent转染液+miRNA-21-5p mimics+光损伤)、阴性组(TransIntro EL Transfection Reagent转染液+miRNA-21-5p mimics NC+光损伤)、PI3K/Akt阻断剂组(TransIntro EL Transfection Reagent转染液+miRNA-21-5p mimics+LY294002+光损伤)。使用光照强度为(16500±200)lx的LED冷光灯建立ARPE-19细胞光损伤模型,利用TransIntro EL Transfection Reagent转染液行细胞转染。采用qRT-PCR法检测各组ARPE-19细胞miRNA-21-5p表达水平,采用CCK-8法检测各组ARPE-19细胞活力,流式细胞仪检测各组ARPE-19细胞活性氧(ROS)含量变化,ELISA法检测各组ARPE-19细胞超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。结果与对照组相比,损伤组ARPE-19细胞miRNA-21-5p表达明显下降,细胞存活率明显下降,ROS含量显著升高,SOD活性明显降低,MDA含量明显增加(均为P<0.001);与损伤组相比,过表达组ARPE-19细胞miRNA-21-5p表达明显升高,细胞存活率明显上升,ROS含量明显降低,SOD活性升高,MDA含量减少(均为P<0.001),而阴性组ARPE-19细胞miRNA-21-5p表达、细胞存活率、ROS含量、SOD活性、MDA含量均无明显差异(均为P>0.05);与过表达组相比,PI3K/Akt阻断剂组ARPE-19细胞miRNA-21-5p表达明显降低,细胞存活率明显下降,ROS含量明显升高,SOD活性明显降低,MDA含量明显增加(均为P<0.01)。结论miRNA-21-5p能显著降低光诱导的ARPE-19细胞氧化应激水平,提高光诱导的ARPE-19细胞抗氧化能力。展开更多
基金Supported by the Nature Science Foundation of Hebei Province,No.H2023104011.
文摘BACKGROUND Diabetic kidney disease(DKD),characterized by increased urinary microalbumin levels and decreased renal function,is the primary cause of end-stage renal di-sease.Its pathological mechanisms are complicated and multifactorial;Therefore,sensitive and specific biomarkers are needed.Urinary exosome originate from diverse renal cells in nephron segments and partially mirror the pathological changes in the kidney.The microRNAs(miRNAs)in urinary exosome are remark-ably stable and highly tissue-specific for the kidney.METHODS Type 2 diabetic mellitus(T2DM)patients were recruited from the Second Hospital of Hebei Medical University and were divided into two groups:DM,diabetic pa-tients without albuminuria[urinary albumin to creatinine ratio(UACR)<30 mg/g]and DKD,diabetic patients with albuminuria(UACR≥30 mg/g).Healthy subjects were the normal control(NC)group.Urinary exosomal miR-145-5p,miR-27a-3p,and miR-29c-3p,were detected using real-time quantitative polymerase chain reaction.The correlation between exosomal miRNAs and the clinical in-dexes was evaluated.The diagnostic values of exosomal miR-145-5p and miR-27a-3p in DKD were determined using receiver operating characteristic(ROC)analysis.Biological functions of miR-145-5p were investigated by performing RESULTS Urinary exosomal expression of miR-145-5p and miR-27a-3p was more upregulated in the DKD group than in the DM group(miR-145-5p:4.54±1.45 vs 1.95±0.93,P<0.001;miR-27a-3p:2.33±0.79 vs 1.71±0.76,P<0.05)and the NC group(miR-145-5p:4.54±1.45 vs 1.55±0.83,P<0.001;miR-27a-3p:2.33±0.79 vs 1.10±0.51,P<0.001).The exosomal miR-145-5p and miR-27a-3p positively correlated with albuminuria and serum creatinine and negatively correlated with the estimated glomerular filtration rate.miR-27a-3p was also closely related to blood glucose,gly-cosylated hemoglobin A1c,and low-density lipoprotein cholesterol.ROC analysis revealed that miR-145-5p had a better area under the curve of 0.88[95%confidence interval(CI):0.784-0.985,P<0.0001]in diagnosing DKD than miR-27a-3p with 0.71(95%CI:0.547-0.871,P=0.0239).Bioinformatics analysis revealed that the target genes of miR-145-5p were located in the actin filament,cytoskeleton,and extracellular exosome and were involved in the pathological processes of DKD,including apoptosis,inflammation,and fibrosis.CONCLUSION Urinary exosomal miR-145-5p and miR-27a-3p may serve as novel noninvasive diagnostic biomarkers or promising therapeutic targets for DKD.
文摘目的研究miRNA-21-5p对光诱导的人视网膜色素上皮细胞氧化应激损伤的影响。方法将体外培养的人视网膜色素上皮细胞系ARPE-19细胞随机分为对照组(TransIntro EL Transfection Reagent转染液培养)、损伤组(TransIntro EL Transfection Reagent转染液+光损伤)、过表达组(TransIntro EL Transfection Reagent转染液+miRNA-21-5p mimics+光损伤)、阴性组(TransIntro EL Transfection Reagent转染液+miRNA-21-5p mimics NC+光损伤)、PI3K/Akt阻断剂组(TransIntro EL Transfection Reagent转染液+miRNA-21-5p mimics+LY294002+光损伤)。使用光照强度为(16500±200)lx的LED冷光灯建立ARPE-19细胞光损伤模型,利用TransIntro EL Transfection Reagent转染液行细胞转染。采用qRT-PCR法检测各组ARPE-19细胞miRNA-21-5p表达水平,采用CCK-8法检测各组ARPE-19细胞活力,流式细胞仪检测各组ARPE-19细胞活性氧(ROS)含量变化,ELISA法检测各组ARPE-19细胞超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。结果与对照组相比,损伤组ARPE-19细胞miRNA-21-5p表达明显下降,细胞存活率明显下降,ROS含量显著升高,SOD活性明显降低,MDA含量明显增加(均为P<0.001);与损伤组相比,过表达组ARPE-19细胞miRNA-21-5p表达明显升高,细胞存活率明显上升,ROS含量明显降低,SOD活性升高,MDA含量减少(均为P<0.001),而阴性组ARPE-19细胞miRNA-21-5p表达、细胞存活率、ROS含量、SOD活性、MDA含量均无明显差异(均为P>0.05);与过表达组相比,PI3K/Akt阻断剂组ARPE-19细胞miRNA-21-5p表达明显降低,细胞存活率明显下降,ROS含量明显升高,SOD活性明显降低,MDA含量明显增加(均为P<0.01)。结论miRNA-21-5p能显著降低光诱导的ARPE-19细胞氧化应激水平,提高光诱导的ARPE-19细胞抗氧化能力。
