Transglutaminase participates in UVB-induced cell death pathways in human corneal epithelial cells.

Transglutaminase participates in UVB-induced cell death pathways in human corneal epithelial cells.
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转谷氨酰胺酶参与 UVB 诱导的人角膜上皮细胞细胞死亡途径。

DOI:
10.1167/iovs.06-0412
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发表时间:
2006
期刊:
Investigative ophthalmology & visual science.
影响因子:
--
通讯作者:
Pflugfelder,StephenC
Pflugfelder,StephenC
中科院分区:
--
文献类型:
--
作者:
Tong,Louis;Chen,Zhuo;DePaiva,CintiaS;Beuerman,Roger;Li,De-Quan;Pflugfelder,StephenC

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目的。已知紫外线 (UVB) 会导致人角膜上皮细胞凋亡。本研究评估了转谷氨酰胺酶在调节肿瘤坏死因子 (TNF) 受体簇以及 UVB 诱导的人角膜上皮细胞凋亡中半胱天冬酶激活中的作用。使用人角膜上皮细胞系。使用单剂量 UVB (20 mJ/cm 2) 作为刺激。通过 MTT、末端脱氧核苷酸转移酶介导的 dUTP-地高辛缺口末端标记 (TUNEL) 和 caspase-3 测定来研究细胞活力和细胞死亡。免疫荧光染色用于研究 UVB 后不同时间间隔的 TNF 受体-I 聚类。使用短干扰 RNA 来降低转谷氨酰胺酶 2 的表达。使用荧光素-尸胺摄取来评估转谷氨酰胺酶活性。使用非共价肽递送系统将豚鼠肝转谷氨酰胺酶转染至角膜上皮细胞。结果。 UVB 增加转谷氨酰胺酶活性,降低细胞活力,并增加 TUNEL 染色。 UVB 或 TNF-α 促进 TNF-受体-I 聚集,这一过程被转谷氨酰胺酶抑制剂单丹酰尸胺抑制。 UVB 还可以通过单丹磺酰尸胺抑制的方式增加活化的 caspase-3。与载体对照相比,细胞内递送外源转谷氨酰胺酶显着增加 caspase-3 活化。结论。转谷氨酰胺酶活性参与 UVB 后角膜上皮细胞死亡,并且似乎参与调节该过程的两个步骤:TNF 受体-I 的聚集和 caspase-3 的激活。
purpose. Ultraviolet light (UVB) is known to cause apoptosis in human corneal epithelial cells. This study evaluates the role of transglutaminase in regulating tumor necrosis factor (TNF) receptor clustering as well as caspase activation in UVB-induced apoptosis in human corneal epithelial cells.methods. A human corneal epithelial cell line was used. A single dose of UVB (20 mJ/cm 2) was used as a stimulus. Cell viability and cell death were investigated by MTT, terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL), and caspase-3 assays. Immunofluorescent staining was used to investigate TNF receptor-I clustering at various time intervals after UVB. Short interfering RNA was used to knock down transglutaminase-2 expression. Fluorescein-cadaverine uptake was used to assess transglutaminase activity. A noncovalent peptide delivery system was used to transfect guinea pig liver transglutaminase into corneal epithelial cells.results. UVB increased transglutaminase activity, reduced cell viability, and increased TUNEL staining. UVB or TNF-α promoted TNF-receptor-I clustering, a process inhibited by the transglutaminase inhibitor, mono-dansyl cadaverine. UVB also increased activated caspase-3, in a manner suppressible by mono-dansyl cadaverine. Intracellular delivery of exogenous transglutaminase markedly increase caspase-3 activation compared with the vehicle control.conclusions. Transglutaminase enzymatic activity is involved in corneal epithelial cell death after UVB and appears to participate in two steps regulating this process, clustering of TNF receptor-I and caspase-3 activation.
维生素 A 缺乏大鼠角膜上角质形成细胞转谷氨酰胺酶的表达。
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