title: ERM protein moesin is phosphorylated by advanced glycation end products and modulates endothelial permeability.
abstract: Advanced glycation end products (AGEs) accumulated in different pathological conditions have the potent capacity to alter cellular properties that include endothelial structural and functional regulations. The disruption of endothelial barrier integrity may contribute to AGE-induced microangiopathy and macrovasculopathy. Previous studies have shown that AGEs induced the rearrangement of actin and subsequent hyperpermeability in endothelial cells (ECs). However, the mechanisms involved in this AGE-evoked EC malfunction are not well understood. This study directly evaluated the involvement of moesin phosphorylation in AGE-induced alterations and the effects of the RhoA and p38 MAPK pathways on this process. Using immortalized human dermal microvascular ECs (HMVECs), we first confirmed that the ezrin/radixin/moesin (ERM) protein moesin is required in AGE-induced F-actin rearrangement and hyperpermeability responses in ECs by knockdown of moesin protein expression with small interfering RNA. We then detected AGE-induced moesin phosphorylation by Western blot analysis. The mechanisms involved in moesin phosphorylation were analyzed by blocking AGE receptor binding and inhibiting Rho and MAPK pathways. AGE-treated HMVECs exhibited time- and dose-dependent increases in the Thr(558) phosphorylation of moesin. The increased moesin phosphorylation was attenuated by preadministrations of AGE receptor antibody, Rho kinase (ROCK), or p38 inhibitor. Suppression of p38 activation via the expression of dominant negative mutants with Ad.MKK6b or Ad.p38alpha also decreased moesin phosphorylation. The activation of the p38 pathway by transfection of HMVECs with an adenoviral construct of dominant active MKK6b resulted in moesin phosphorylation. These results suggest a critical role of moesin phosphorylation in AGE-induced EC functional and morphological regulations. Activation of the ROCK and p38 pathways is required in moesin phosphorylation.
authors.affiliation: Department of Pathophysiology, Southern Medical University, Guangzhou, PR China.
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Guo
authors.forename: Lingjun
authors.initials: L
authors.lastname: Wang
authors.forename: Bo
authors.initials: B
authors.lastname: Chen
authors.forename: Qiang
authors.initials: Q
authors.lastname: Li
authors.forename: Jiping
authors.initials: J
authors.lastname: Wang
authors.forename: Ming
authors.initials: M
authors.lastname: Zhao
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
authors.forename: Xuliang
authors.initials: X
authors.lastname: Huang
authors.forename: Qiaobing
authors.initials: Q
authors.lastname: Huang
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D002199:Capillary Permeability / Q000502:physiology; D002478:Cells, Cultured; D003198:Computer Simulation; D003599:Cytoskeleton / Q000502:physiology; D004730:Endothelium, Vascular / Q000502:physiology*; D005456:Fluorescent Dyes; D005822:Genetic Vectors; D017127:Glycation End Products, Advanced / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D010766:Phosphorylation; D034741:RNA, Small Interfering; D015398:Signal Transduction / Q000502:physiology; D048051:p38 Mitogen-Activated Protein Kinases / Q000502:physiology; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D017127:Glycation End Products, Advanced; D008840:Microfilament Proteins; D034741:RNA, Small Interfering; C071058:moesin; D054460:rho-Associated Kinases; D048051:p38 Mitogen-Activated Protein Kinases