title: IP3 and calcium signaling involved in the reorganization of the actin cytoskeleton and cell rounding induced by cigarette smoke extract in human endothelial cells.
abstract: Smoking increases the risk of cardiovascular disorders and leads to damage caused by inflammation and oxidative stress. The actin cytoskeleton is a key player in the response to inflammatory stimuli and is an early target of cellular oxidative stress. The purpose of this study was to investigate the changes in actin cytoskeleton dynamics in human endothelial EA.hy926 cells exposed to cigarette smoke extract (CSE). Immunostaining revealed that CSE exposure resulted in modification of the actin cytoskeleton and led to cell rounding in a dose- and time-dependent manner. In addition, the intracellular calcium concentration was increased by treatment with CSE. Pretreatment with antioxidants (lipoic acid, glutathione, N-acetyl cysteine, aminoguanidine, α-tocopherol, and vitamin C) significantly attenuated the CSE-induced actin cytoskeleton reorganization and cell rounding. Calcium ion chelators (EGTA, BAPTA-AM AM) and a potent store-operated calcium channel inhibitor (MRS 1845) also reduced CSE-induced intracellular calcium changes and attenuated actin cytoskeleton reorganization and cell morphology change. Moreover, the CSE-induced intracellular calcium increase was suppressed by pretreatment with the inositol trisphosphate receptor (IP3R) inhibitor xestospongin C, the phospholipase C (PLC) inhibitor U-73122, and the protein kinase C (PKC) inhibitor GF109203X. These results suggest that reactive oxygen species production and intracellular calcium increase play an essential role in CSE-induced actin disorganization and cell rounding through a PLC-IP3-PKC signaling pathway. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1293-1306, 2016.
authors.affiliation: School of Biomedical Sciences, Chung Shan Medical University, Taichung, Taiwan.
authors.forename: Bo-Hong
authors.initials: BH
authors.lastname: Lin
authors.affiliation: School of Biomedical Sciences, Chung Shan Medical University, Taichung, Taiwan.
authors.forename: Ming-Hsuan
authors.initials: MH
authors.lastname: Tsai
authors.affiliation: Department of Nutrition, China Medical University, Taichung, Taiwan.|Department of Health and Nutrition Biotechnology, Asia University, Taichung, Taiwan.
authors.forename: Chong-Kuei
authors.initials: CK
authors.lastname: Lii
authors.affiliation: School of Biomedical Sciences, Chung Shan Medical University, Taichung, Taiwan. tsushing@csmu.edu.tw.|Department of Medical Research, Chung Shan Medical University Hospital, Taichung, Taiwan. tsushing@csmu.edu.tw.
authors.forename: Tsu-Shing
authors.initials: TS
authors.lastname: Wang
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism*; D000975:Antioxidants / Q000494:pharmacology; D020013:Calcium Signaling / Q000187:drug effects; D002460:Cell Line; D002614:Chelating Agents / Q000494:pharmacology; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004963:Estrenes / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000037:antagonists & inhibitors / Q000378:metabolism*; D047028:Macrocyclic Compounds / Q000494:pharmacology; D008301:Maleimides / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D009568:Nitrendipine / Q000031:analogs & derivatives / Q000494:pharmacology; D010080:Oxazoles / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D011760:Pyrrolidinones / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D012906:Smoke; D014026:Nicotiana / Q000737:chemistry* / Q000378:metabolism; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D000975:Antioxidants; D002614:Chelating Agents; D004963:Estrenes; D007211:Indoles; D053496:Inositol 1,4,5-Trisphosphate Receptors; C503978:MRS 1845; D047028:Macrocyclic Compounds; D008301:Maleimides; D010080:Oxazoles; D011760:Pyrrolidinones; D017382:Reactive Oxygen Species; D012906:Smoke; C511704:xestospongin C; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; D009568:Nitrendipine; D011493:Protein Kinase C; D010738:Type C Phospholipases; D005978:Glutathione; C070515:bisindolylmaleimide I; D000111:Acetylcysteine
keywords: actin cytoskeleton reorganization; calcium signal; cigarette smoke extract; human endothelial EA.hy926 cells; inositol trisphosphate receptor; phospholipase C; protein kinase C