Sirt1 Regulates Corneal Epithelial Migration by Deacetylating Cortactin.
Sirt1 Regulates Corneal Epithelial Migration by Deacetylating Cortactin.
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Sirt1 通过去乙酰 Cortactin 调节角膜上皮迁移
DOI：
10.1167/iovs.63.12.14
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发表时间：
2022-11-01
影响因子：
4.4
通讯作者：
中科院分区：
文献类型：
--
作者：
目的沉默信息调节因子1（SIRT 1）是一种依赖于NAD+的脱乙酰酶，在细胞代谢、自噬和染色质可及性中起重要作用。我们的研究旨在确定其在控制角膜上皮伤口愈合（CEWH）中的作用。方法采用Cre-lox系统建立角膜上皮特异性Sirt 1基因缺失小鼠模型。使用CE清创术创建CEWH模型。用Algerbrush收集角膜上皮细胞（CEC）。进行蛋白质印迹分析和RT-qPCR以确定蛋白质和mRNA表达水平。SiRNA转染技术可降低人角膜上皮细胞SIRT 1和coronin的表达水平。划痕试验、MTS试验和TUNEL试验分别测定细胞迁移、增殖和凋亡行为。免疫共沉淀法检测SIRT 1和coronin的相互作用。免疫荧光染色评价SIRT 1、coronin、acetylated-coronin和F-actin的位置和表达水平。结果在CEWH过程中，SIRT 1 mRNA和蛋白表达水平的增加伴随着非组蛋白中乙酰化赖氨酸的下调。SIRT 1功能的丧失减少了细胞迁移，进而延迟了CEWH。SIRT 1在体外和体内结合和脱乙酰化coronin。SIRT 1或corneum蛋白的缺失抑制了伤口边缘片状伪足的形成，这与迁移阻滞一致。结论在CEWH过程中，SIRT 1上调及其对coronin的修饰通过增加伤口边缘板状伪足的发育来促进CEC迁移。因此，SIRT 1可以作为增强CEWH的潜在靶点。
Purpose Silent information regulator 1 (SIRT1) is a nicotinamide adenine dinucleotide (NAD+) dependent deacetylase, which plays an essential role in cellular metabolism, autophagy, and chromatin accessibility. Our study aimed to determine its role in controlling corneal epithelial wound healing (CEWH). Methods Corneal epithelial (CE)–specific Sirt1 deletion mice were created using the Cre-lox system. CE debridement was used to create a CEWH model. Corneal epithelial cells (CECs) were collected with an Algerbrush. Western blot analysis and RT-qPCR were performed to determine protein and mRNA expression levels. SiRNA transfection technology knocked down SIRT1 and cortactin expression levels in human corneal epithelial cells. Scratch wound assay, MTS assay, and TUNEL assay determined cell migratory, proliferative, and apoptotic behavior, respectively. Co-immunoprecipitation probed for SIRT1 and cortactin interaction. Immunofluorescence staining evaluated the location and expression levels of SIRT1, cortactin, acetylated-cortactin, and F-actin. Results During CEWH, increases in SIRT1 mRNA and protein expression levels accompanied the downregulation of acetylated lysine in non-histone proteins. The loss of SIRT1 function reduced cell migration and, in turn, delayed CEWH. SIRT1 bound to and deacetylated cortactin in vitro and in vivo. Loss of either SIRT1 or cortactin suppressed wound edge lamellipodia formation, which is consistent with migration retardation. Conclusions During CEWH, SIRT1 upregulation and its modification of cortactin boost CEC migration by increasing the development of lamellipodia at the wound edge. Therefore SIRT1 may serve as a potential target to enhance CEWH.
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影响因子：
56.9
作者：
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通讯作者：
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通讯作者：
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影响因子：
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作者：
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影响因子：
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