Zhang Junfang, Luo Jiawei, Zhang Guowei, Qin Bai, Ren Xiumei, Guan Huaijin
Eye Institute, Affiliated Hospital of Nantong University, Nantong, China.
Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai, China.
J Ophthalmol. 2022 Apr 21;2022:6706172. doi: 10.1155/2022/6706172. eCollection 2022.
To identify the ability of releasing and uptaking exosomes in rat lens and characterize the exosomal microRNA profile of lens-derived exosomes.
The rat lenses were cultured ex vivo and the medium was collected. The exosomes were isolated from medium and measured in size and concentration by nanoflow cytometry (nFCM) and transmission electron microscopy (TEM) and verified with CD63 and TSG101 by Western blot. The miRNAs in exosomes released from lens epithelial cells (LECs) were sequenced. The plasma exosomes labeled by PKH26 were used to verify the exosomes uptake LECs, and their colocalized fluorescence was imaged by confocal microscopy.
LECs released numerous exosomes into the medium through the capsule, which contained abundant miRNAs. The most abundant miRNAs included miR-184, let-7c-5p, let-7a-5p, let-7b-5p, let-7f-5p, miR-125a-5p, miR-204-5p, miR-125b-5p, miR-1b, and miR-23a-3p. The LECs but not the lens fibre cells showed exosome uptake. The LECs uptake more PKH26-labeled exosomes at day 7 than day 3 and day 14.
Our results suggested that LECs can release and uptake exosomes through the capsule. Exosomes may be an important way for the lens to communicate among LECs, aqueous humour, vitreous body, and other ocular tissues.
鉴定大鼠晶状体释放和摄取外泌体的能力，并表征晶状体来源外泌体的微小RNA谱。
对大鼠晶状体进行离体培养并收集培养基。从培养基中分离出外泌体，通过纳米流式细胞术（nFCM）和透射电子显微镜（TEM）测量其大小和浓度，并通过蛋白质免疫印迹法用CD63和TSG101进行验证。对晶状体上皮细胞（LECs）释放的外泌体中的微小RNA进行测序。用PKH26标记的血浆外泌体来验证LECs对外泌体的摄取，并用共聚焦显微镜对其共定位荧光进行成像。
LECs通过晶状体囊将大量外泌体释放到培养基中，这些外泌体含有丰富的微小RNA。最丰富的微小RNA包括miR-184、let-7c-5p、let-7a-5p、let-7b-5p、let-7f-5p、miR-125a-5p、miR-204-5p、miR-125b-5p、miR-1b和miR-23a-3p。只有LECs而非晶状体纤维细胞表现出对外泌体的摄取。LECs在第7天比第3天和第14天摄取更多PKH26标记的外泌体。
我们的结果表明，LECs可通过晶状体囊释放和摄取外泌体。外泌体可能是晶状体在LECs、房水、玻璃体和其他眼组织之间进行通讯的重要方式。
