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Exosomal miR-29b found in aqueous humour mediates calcium signaling in diabetic patients with cataract

查看全文 作　　者：Chao [1,2]Gao;Xin [2,3,4,5]Liu;Fan [2,3,4,5]Fan;Jia-Ning [2,3,4,5]Yang;Xi-Yue [2,3,4,5]Zhou;Heng-Jun [2,3,4,5]Mei;Xiao-Lei [2,3,4,5]Lin;Yi [2,3,4,5]Luo 高影响力作者 机构地区：[1]First Affiliated Hospital,School of Medicine,Shihezi University,Shihezi 832000,Xinjiang Uygur Autonomous Region,China;[2]Eye Institute,Eye and ENT Hospital,College of Medicine,Fudan University,Shanghai 200031,China;[3]State Key Laboratory of Medical Neurobiology,Institutes of Brain Science and Collaborative Innovation Center for Brain Science,Shanghai Medical College,Fudan University,Shanghai 200031,China;[4]Shanghai Key Laboratory of Visual Impairment and Restoration,Science and Technology Commission of Shanghai Municipality,Shanghai 200031,China;[5]Key Laboratory of Myopia(Fudan University),Chinese Academy of Medical Sciences,National Health Commission,Shanghai 200031,China高影响力机构 出　　处：《International Journal of Ophthalmology(English edition)》索引2021年第14卷第10期,共8页高影响力期刊 基　　金：the National Natural Science Foundation of China(No.81870645)。 摘　　要：AIM:To investigate the role of exosomal miR-29b and Ca^(2+)in regulating the function of human lens epithelial cells(HLECs).METHODS:Exosomes were isolated from human aqueous humour(AH)by ultracentrifugation,and visualized by nanopar ticle tracking and transmission electron microscopy.Exosomal miRNA sequencing was performed to identify differentially expressed miRNAs between diabetes with cataracts(DMC)group and age-related cataracts(ARC)group.TargetScan was used to predict potential target of certain miRNA.The expression of CACNA1C mRNA was determined by quantitative real-time polymerase chain reaction and CACNA1C protein was determined by Western blotting.Concentration of Ca^(2+)in human AH and the culture supernatant of cells were detected by the calcium assay kit.Cell counting kit-8 was used to determine cell viability.RESULTS:Exosomes were isolated from human AH,which had a typical cup-shaped phenotype and a particle size distribution in accordance with micro extracellular vesicles.Exosomal miRNA sequencing revealed that miR-29b was significantly downregulated in DMC group compared with ARC.Ca^(2+)concentration of human AH in DMC was higher than that in ARC.The culture supernatant of cells transfected with miR-29b inhibitors had a higher concentration of Ca^(2+)than that transfected with miR-29b mimics.miR-29b reduced the viability of HLECs by upregulating CACNA1C expression.CONCLUSION:Exosomes isolated from human AH contains abundant miRNAs.A significantly expressed miR NA,miR-29b,can affect the concentration of Ca^(2+)and regulate HLEC processes by upregulating CACNA1C. 关 键 词：EXOSOMES miR-29b diabetes and cataracts CA^(2+) CACNA1C
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