4-HNE Induces Apoptosis of Human Retinal Pigment Epithelial Cells by Modifying HSP70

• Published in: Current medical science Vol. 39; no. 3; pp. 442 - 448
• Main Authors: Yang, Lei-lei, Chen, Hao, Wang, Jun, Xia, Ting, Sun, Hong, Yuan, Chun-hui, Liu, Shi-liang, Chen, Jian-bin
• Format: Journal Article
• Language: English
• Published: Wuhan Huazhong University of Science and Technology 01.06.2019
• Subjects: Aldehydes - pharmacology; Apoptosis - drug effects; Apoptosis - genetics; bcl-2-Associated X Protein - genetics; bcl-2-Associated X Protein - metabolism; Cell Line; Epithelial Cells - cytology; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Gene Expression Regulation; HSP70 Heat-Shock Proteins - chemistry; HSP70 Heat-Shock Proteins - genetics; HSP70 Heat-Shock Proteins - metabolism; Humans; Lipid Peroxidation; Medicine; Medicine & Public Health; Plasmids - chemistry; Plasmids - metabolism; Protein Binding; Proto-Oncogene Proteins c-bcl-2 - genetics; Proto-Oncogene Proteins c-bcl-2 - metabolism; Retinal Pigments - metabolism; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; Signal Transduction; Transfection; X-Linked Inhibitor of Apoptosis Protein - antagonists & inhibitors; X-Linked Inhibitor of Apoptosis Protein - genetics; X-Linked Inhibitor of Apoptosis Protein - metabolism; human retinal pigment epithelial cells; XIAP; apoptosis; HSP70; 4-hydroxy-2-nonenal
• ISSN: 2096-5230; 2523-899X
• DOI: 10.1007/s11596-019-2057-8

Summary The role of heat shock protein 70 (HSP70) in apoptosis of human retinal pigment epithelial cells (ARPE-19) induced by 4-hydroxy-2-nonenal (4-HNE) was explored. Different concentrations of 4-HNE were used to stimulate ARPE-19 cells, and apoptosis was measured by flow cytometry. The expression of apoptotic-related proteins, HSP70, X-linked inhibitor-of-apoptosis (XIAP), Bcl-2, and Bax were quantified by Western blotting. HSP70 and XIAP overexpression plasmids, or their corresponding siRNAs were transfected into ARPE-19 cells using Lipofectamine™ 2000. Co-immunoprecipitation and Western blotting were used to detect the effect of 4-HNE on the expression of HSP70 and the binding level between 4-HNE and HSP70. The results showed that 4-HNE induced late apoptosis in ARPE-19 cells, accompanied by elevated levels of 4-HNE-modified HSP70, but it did not affect HSP70 protein expression. 4-HNE-modified HSP70 down-regulated the expression of the apoptosis inhibitory protein XIAP. Overexpression of HSP70 or XIAP inhibited 4-HNE-induced apoptosis of ARPE-19 cells. It was suggested that 4-HNE could promote XIAP degradation by modification of HSP70 to induce late apoptosis of human retinal pigment epithelial cells.