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
Cysteine-rich secretory protein 2 （CRISP2） is an important protein in spermatozoa that plays roles in modulating sperm flagellar motility, the acrosome reaction, and gamete fusion. Spermatozoa lacking CRISP2 exhibit...Cysteine-rich secretory protein 2 （CRISP2） is an important protein in spermatozoa that plays roles in modulating sperm flagellar motility, the acrosome reaction, and gamete fusion. Spermatozoa lacking CRISP2 exhibit low sperm motility and abnormal morphology. However, the molecular mechanisms underlying the reduction of CRISP2 in asthenoteratozoospermia （ATZ） remain unknown. In this study, low expression of CRISP2 protein rather than its mRNA was observed in the ejaculated spermatozoa from ATZ patients as compared with normozoospermic males. Subsequently, bioinformatic prediction, luciferase reporter assays, and microRNA-27a （miR-27a） transfection experiments revealed that miR-27a specifically targets CRISP2 by binding to its 3＇ untranslated region （3＇-UTR）, suppressing CRISP2 expression posttranscriptionally. Further evidence was provided by the clinical observation of high miR-27a expression in ejaculated spermatozoa from ATZ patients and a negative correlation between miR-27a expression and CRISP2 protein expression. Finally, a retrospective follow-up study supported that both high miR-27a expression and low CRISP2 protein expression were associated with low progressive sperm motility, abnormal morphology, and infertility. This study demonstrates a novel mechanism responsible for reduced CRISP2 expression in ATZ, which may offer a potential therapeutic target for treating male infertility, or for male contraception.展开更多
目的探讨人微小RNA-27a(microRNA-27a,miR-27a)及其靶基因FOXO1在胃癌组织、癌旁组织中的变化及其在金雀异黄素抑制胃癌细胞增殖中的作用。方法利用Real Time PCR方法检测10对胃癌癌旁/癌组织中microRNA-27a及其靶基因FOXO1mRNA的表达...目的探讨人微小RNA-27a(microRNA-27a,miR-27a)及其靶基因FOXO1在胃癌组织、癌旁组织中的变化及其在金雀异黄素抑制胃癌细胞增殖中的作用。方法利用Real Time PCR方法检测10对胃癌癌旁/癌组织中microRNA-27a及其靶基因FOXO1mRNA的表达。不同浓度(5、10、20、40μmol.L-1)的金雀异黄素作用于胃癌细胞SGC-7901 72h后,用噻唑兰比色法(MTT)检测细胞增殖,并采用Real Time PCR方法检测细胞中miR-27a及其靶基因FOXO1mRNA的表达。结果胃癌组织较癌旁组织miR-27a表达升高(1.566±0.206 5),其靶基因FOXO1mRNA表达降低(0.298±0.156 6)。Pearson线性分析表明:miR-27a与FOXO1mRNA呈负相关(r=-0.701 8)。金雀异黄素作用于SGC-7901细胞后,细胞增殖得到了抑制,尤其在20与40μmol.L-1浓度下,细胞抑制率明显降低。不同浓度的金雀异黄素处理72h后,miR-27a表达与对照组比较明显降低;其靶基因FOXO1mRNA表达水平较对照组增加(均P<0.05)。结论金雀异黄素可能通过抑制miR-27a表达,促进FOXO1mR-NA的表达,进而抑制胃癌细胞增殖,发挥抗肿瘤的作用。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.
基金The authors would like to thank Prof. Chunyan Wang, for her contributions to the samples collection, as well as thank Cheng Yang for his contributions to the figures revision. This study was supported by the Guangdong Provincial Natural Science Foundation of China （No. 2014A030313291, No. 2015A030310027 and No. 2016A030310393）, the Guangdong Provincial Science and Technology Program （No. 2014A020212204）, and the Science and Technology Innovation Project of Southern Medical University （QD2014N005）.
文摘Cysteine-rich secretory protein 2 （CRISP2） is an important protein in spermatozoa that plays roles in modulating sperm flagellar motility, the acrosome reaction, and gamete fusion. Spermatozoa lacking CRISP2 exhibit low sperm motility and abnormal morphology. However, the molecular mechanisms underlying the reduction of CRISP2 in asthenoteratozoospermia （ATZ） remain unknown. In this study, low expression of CRISP2 protein rather than its mRNA was observed in the ejaculated spermatozoa from ATZ patients as compared with normozoospermic males. Subsequently, bioinformatic prediction, luciferase reporter assays, and microRNA-27a （miR-27a） transfection experiments revealed that miR-27a specifically targets CRISP2 by binding to its 3＇ untranslated region （3＇-UTR）, suppressing CRISP2 expression posttranscriptionally. Further evidence was provided by the clinical observation of high miR-27a expression in ejaculated spermatozoa from ATZ patients and a negative correlation between miR-27a expression and CRISP2 protein expression. Finally, a retrospective follow-up study supported that both high miR-27a expression and low CRISP2 protein expression were associated with low progressive sperm motility, abnormal morphology, and infertility. This study demonstrates a novel mechanism responsible for reduced CRISP2 expression in ATZ, which may offer a potential therapeutic target for treating male infertility, or for male contraception.
文摘目的探讨人微小RNA-27a(microRNA-27a,miR-27a)及其靶基因FOXO1在胃癌组织、癌旁组织中的变化及其在金雀异黄素抑制胃癌细胞增殖中的作用。方法利用Real Time PCR方法检测10对胃癌癌旁/癌组织中microRNA-27a及其靶基因FOXO1mRNA的表达。不同浓度(5、10、20、40μmol.L-1)的金雀异黄素作用于胃癌细胞SGC-7901 72h后,用噻唑兰比色法(MTT)检测细胞增殖,并采用Real Time PCR方法检测细胞中miR-27a及其靶基因FOXO1mRNA的表达。结果胃癌组织较癌旁组织miR-27a表达升高(1.566±0.206 5),其靶基因FOXO1mRNA表达降低(0.298±0.156 6)。Pearson线性分析表明:miR-27a与FOXO1mRNA呈负相关(r=-0.701 8)。金雀异黄素作用于SGC-7901细胞后,细胞增殖得到了抑制,尤其在20与40μmol.L-1浓度下,细胞抑制率明显降低。不同浓度的金雀异黄素处理72h后,miR-27a表达与对照组比较明显降低;其靶基因FOXO1mRNA表达水平较对照组增加(均P<0.05)。结论金雀异黄素可能通过抑制miR-27a表达,促进FOXO1mR-NA的表达,进而抑制胃癌细胞增殖,发挥抗肿瘤的作用。
