Biological effects of cigarette smoke in cultured human retinal pigment epithelial cells

• Published in: PloS one 2012-11, Vol.7 (11), p.e48501-e48501
• Main Authors: Yu, Alice L, Birke, Kerstin, Burger, Johannes, Welge-Lussen, Ulrich
• Format: Article
• Language: English
• Subjects: Adult; Age; Analysis; Apolipoproteins; Apoptosis; beta-Galactosidase - metabolism; Biological effects; Biology; Cell culture; Cell death; Cell Death - drug effects; Cell Survival - drug effects; Cells, Cultured; Cellular Senescence - drug effects; Cigarette smoke; Cigarettes; Clusterin - metabolism; Connective tissue growth factor; Connective Tissue Growth Factor - metabolism; Connective tissues; Disease; Enzyme-linked immunosorbent assay; Exposure; Extracellular matrix; Fibronectin; Fibronectins; Fibronectins - metabolism; Fluorescence; Galactosidase; Gene expression; Histology; Humans; Laminin; Laminin - metabolism; Lipid peroxidation; Lipid Peroxidation - drug effects; Macular degeneration; Medicine; Oxidative stress; Pathogenesis; Peptide nucleic acids; Peroxidation; Protein folding; Retina; Retinal pigment epithelium; Retinal Pigment Epithelium - cytology; Retinal Pigment Epithelium - drug effects; Retinal Pigment Epithelium - metabolism; RNA; Secretion; Senescence; Smoke; Smoking; Smoking - adverse effects; Synthesis
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0048501

The goal of the present study was to determine whether treatment with cigarette smoke extract (CSE) induces cell loss, cellular senescence, and extracellular matrix (ECM) synthesis in primary human retinal pigment epithelial (RPE) cells. Primary cultured human RPE cells were exposed to 2, 4, 8, and 12% of CSE concentration for 24 hours. Cell loss was detected by cell viability assay. Lipid peroxidation was assessed by loss of cis-parinaric acid (PNA) fluorescence. Senescence-associated ß-galactosidase (SA-ß-Gal) activity was detected by histochemical staining. Expression of apolipoprotein J (Apo J), connective tissue growth factor (CTGF), fibronectin, and laminin were examined by real-time PCR, western blot, or ELISA experiments. The results showed that exposure of cells to 12% of CSE concentration induced cell death, while treatment of cells with 2, 4, and 8% CSE increased lipid peroxidation. Exposure to 8% of CSE markedly increased the number of SA-ß-Gal positive cells to up to 82%, and the mRNA expression of Apo J, CTGF, and fibronectin by approximately 3-4 fold. Treatment with 8% of CSE also increased the protein expression of Apo J and CTGF and the secretion of fibronectin and laminin. Thus, treatment with CSE can induce cell loss, senescent changes, and ECM synthesis in primary human RPE cells. It may be speculated that cigarette smoke could be involved in cellular events in RPE cells as seen in age-related macular degeneration.