title: Effects of 5-aza-2'-deoxycytidine and trichostatin A on high glucose- and interleukin-1β-induced secretory mediators from human retinal endothelial cells and retinal pigment epithelial cells.
abstract: OBJECTIVE
  We aimed to elucidate the effects of two epigenetic inhibitors, 5-aza-2'-deoxycytidine (5-aza-dC) and trichostatin A (TSA), on several key secretory mediators of diabetic retinopathy (DR) in human retinal endothelial cells (HRECs) and human retinal pigment epithelial (HRPE) cells treated with high glucose or interleukin-1β (IL-1β).
  
  
  METHODS
  HRECs and HRPE cells were incubated in 30 mM D-glucose or 10 ng/ml IL-1β with or without the presence of various concentrations of 5-aza-dC or TSA. The production of pigment epithelium derived factor (PEDF), vascular endothelial cell growth factor (VEGF), intercellular cell adhesion molecule-1 (ICAM-1), IL-1β, and matrix metalloproteinase 2 (MMP2) was evaluated at the mRNA and protein levels using real-time PCR and enzyme-linked immunosorbent assay (ELISA), respectively.
  
  
  RESULTS
  In the 30 mM D-glucose and the 10 ng/ml IL-1β condition, the expression of VEGF, ICAM-1, IL-1β, and MMP2 was induced in the HRECs and the HRPE cells. PEDF was downregulated in the HRPE cells but upregulated in the HRECs. However, the PEDF-to-VEGF ratio, which is thought to be critical in DR, was downregulated in both cell types. 5-aza-dC dose-dependently alleviated VEGF, ICAM-1, IL-1β, and MMP2 and reversed PEDF or the PEDF/VEGF ratio in both cell types. TSA had similar effects as 5-aza-dC on the target mediators. However, ICAM-1 production was aggravated in the HRECs while remaining unchanged in the HRPE cells after TSA was administered.
  
  
  CONCLUSIONS
  Our results demonstrated that 5-aza-dC and TSA enhance the protective PEDF and the PEDF/VEGF ratio and ameliorate the adverse effects of diabetic stimuli in vitro, suggesting that these two drugs may be of potential therapeutic value in DR.
authors.affiliation: The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
authors.forename: Manyun
authors.initials: M
authors.lastname: Xie
authors.affiliation: The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
authors.forename: Jingyi
authors.initials: J
authors.lastname: Tian
authors.affiliation: The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Luo
authors.affiliation: The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
authors.forename: Liqing
authors.initials: L
authors.lastname: Wei
authors.affiliation: The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
authors.forename: Shaofen
authors.initials: S
authors.lastname: Lin
authors.affiliation: Aier School of Ophthalmology, Central South University, Changsha, China ; Aier Eye Hospital Group, Changsha, China ; Aier Research Institute of Ophthalmology, Changsha, China.
authors.forename: Shibo
authors.initials: S
authors.lastname: Tang
mesh_terms: D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000077209:Decitabine; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005136:Eye Proteins / Q000096:biosynthesis / Q000378:metabolism; D015870:Gene Expression; D005947:Glucose / Q000037:antagonists & inhibitors / Q000494:pharmacology; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D053583:Interleukin-1beta / Q000037:antagonists & inhibitors / Q000494:pharmacology; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D009414:Nerve Growth Factors / Q000096:biosynthesis / Q000378:metabolism; D061251:Primary Cell Culture; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D012160:Retina / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D012168:Retinal Pigments / Q000378:metabolism; D015843:Serpins / Q000096:biosynthesis / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis / Q000378:metabolism; D000099174:Pigment Epithelium-Derived Factor
chemical_list: D000964:Antimetabolites, Antineoplastic; D001374:Azacitidine; D000077209:Decitabine; D005136:Eye Proteins; D005947:Glucose; D006877:Hydroxamic Acids; D018799:Intercellular Adhesion Molecule-1; D053583:Interleukin-1beta; D020778:Matrix Metalloproteinase 2; D009414:Nerve Growth Factors; D012333:RNA, Messenger; D012168:Retinal Pigments; D015843:Serpins; D042461:Vascular Endothelial Growth Factor A; D000099174:Pigment Epithelium-Derived Factor; C467484:VEGFA protein, human; C012589:trichostatin A; C522361:MMP2 protein, human