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Updated: Sep 12, 2025

Isolation of Mouse Epidermal Keratinocytes and Their In Vitro Clonogenic Culture

Isolation of Mouse Epidermal Keratinocytes and Their In Vitro Clonogenic Culture

Published on: August 10, 2019

通过epigallocatechin gallate救援Sirtuin抑制剂依赖的克劳丁-4表达和细胞屏障性质的降低,通过epigallocatechin gallate在角质细胞中

Maika Miwa¹, Miki Tanabe¹, Shunsuke Matsuda¹

¹Laboratory of Biochemistry, Department of Biopharmaceutical Sciences, Gifu Pharmaceutical University, Gifu 501-1196, Japan.

Biochimica et biophysica acta. Biomembranes

|August 7, 2025

概括

老龄化通过降低Sirtuin-2 (SIRT2) 活性来降低皮肤屏障功能,影响Claudin-4 (CLDN4) 表达. 绿茶中的表甲基酸盐 (EGCG) 激活SIRT2,恢复屏障功能,并可能对抗皮肤衰老.

科学领域:

皮肤病学皮肤病学
细胞生物学细胞生物学
生物化学生化学

背景情况:

皮肤屏障功能对保护至关重要,并由紧密结合蛋白维持,如Claudin-1 (CLDN1) 和Claudin-4 (CLDN4).
随着年龄的增长,SIRT2活性在角质细胞中下降,影响CLDN4的表达和皮肤屏障的完整性.

研究的目的:

研究SIRT2在调节CLDN4表达和皮肤屏障功能的作用.
评估SIRT2激活剂,如表甲基酸盐 (EGCG) 在抵消与年龄相关的障碍障碍功能障碍方面的潜力.

主要方法:

采用了体外SIRT2活性测定和人类角质细胞衍生的HaCaT细胞.
使用SIRT2抑制剂Tenovin-1 (Ten-1) 和EGCG或绿茶提取物 (GT) 来评估对CLDN4表达,局部化和乙化的影响.
使用FD4作为流量标记物测量了体电阻和细胞透性.

主要成果:

-1降低了SIRT2活性,CLDN4蛋白水平,CLDN4紧接点局部化,并在K196.4增加了CLDN4乙化.
EGCG和GT挽救了SIRT2活动,并逆转了Ten-1引起的CLDN4表达和屏障功能的下降.
-1通过K196乙化影响了CLDN4表达,而CLDN1表达不受-1或EGCG的影响.

结论:

通过调节CLDN4的表达和局部化,SIRT2在维持皮肤屏障功能方面发挥着关键作用.
通过激活SIRT2和抑制CLDN4乙化,EGCG显示出作为治疗剂的潜力,可以增强皮肤屏障功能并减轻因衰老引起的衰退.

关键词:

衰老的衰老衰老的衰老克劳迪因-4 类药物.紧密的结口紧密的结口.

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