title: Homocysteine-induced hypermethylation of DDAH2 promoter contributes to apoptosis of endothelial cells.
abstract: Homocysteine (Hcy) could induce apoptosis of endothelial cells (ECs). Dimethylarginine dimethylaminohydrolase 2 (DDAH2) is recognized as a protective factor to improve the endothelial function. Defect of DDAH2 has been confirmed to be involved in the Hcy-induced dysfunction of endothelial NO system. This study was to determine whether Hcy could inhibit DDAH2 expression and induce apoptosis of ECs via increasing DNA methylation level of DDAH2 promoter and whether DNA methylation inhibitor 5-azacytidine (5-aza) could attenuate the effect of Hcy on ECs. Firstly, human umbilical vein endothelial cells (HUVECs) were treated by Hcy with or without 5-aza for 48 h. MTT assay showed that Hcy reduced the viability of HUVECs in a dose-dependent manner. The level of asymmetric dimethylarginine (ADMA) and the apoptosis rate of HUVECs treated with Hcy at 1.0 mM were increased significantly compared with that of control. Moreover, we found that mRNA level of DDAH2 was down-regulated and DNA methylation level of DDAH2 promoter was increased significantly in HUVECs treated with Hcy, in concomitance with up-regulated protein level of DNA methyltransferase 1 (DNMT1). Furthermore, we also found that 5-aza could neutralize the effect of Hcy on ECs through up-regulating mRNA level of DDAH2 and inducing demethylation of DDAH2 promoter. In conclusion, hypermethylation of DDAH2 contributes to Hcy induced apoptosis of ECs. Modulating DNA methylation status of DDAH2 promoter may be a potential strategy for treatment of endothelial dysfunction.
authors.affiliation: Department of Pharmaceutics, The Third XiangYa Hospital, Central South University, Changsha, China.
authors.forename: Su-Jie
authors.initials: SJ
authors.lastname: Jia
authors.forename: Yong-Quan
authors.initials: YQ
authors.lastname: Lai
authors.forename: Ming
authors.initials: M
authors.lastname: Zhao
authors.forename: Ting
authors.initials: T
authors.lastname: Gong
authors.forename: Bi-Kui
authors.initials: BK
authors.lastname: Zhang
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism*; D017304:Annexin A5 / Q000378:metabolism; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D001374:Azacitidine / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D004396:Coloring Agents; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism*; D006710:Homocysteine / Q000494:pharmacology*; D008745:Methylation; D011401:Promoter Regions, Genetic / Q000187:drug effects; D060888:Real-Time Polymerase Chain Reaction; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics; D013778:Tetrazolium Salts; D013844:Thiazoles
chemical_list: D017304:Annexin A5; D000964:Antimetabolites, Antineoplastic; D004396:Coloring Agents; C411744:DMAP1 protein, human; D012097:Repressor Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D006710:Homocysteine; D000581:Amidohydrolases; C059999:dimethylargininase; C022616:thiazolyl blue; D001374:Azacitidine