Ad-hVEGF165 reverses homocysteine-induced endothelial dysfunction by regulating nitric oxide system.
- Author:
Lili ZHAO
1
,
2
;
Email: TJZHAOLL@TOM.COM.
;
Yongmin MAO
3
;
Ying ZHANG
3
;
Ying SHI
3
;
Yanqiu SONG
3
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; Amidohydrolases; metabolism; Cells, Cultured; Homocysteine; adverse effects; Human Umbilical Vein Endothelial Cells; drug effects; Humans; Nitric Oxide; metabolism; Nitric Oxide Synthase Type III; metabolism; RNA, Messenger; metabolism; Recombinant Proteins; pharmacology; Vascular Endothelial Growth Factor A; pharmacology
- From: Chinese Journal of Cardiology 2015;43(5):432-436
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the therapeutic effect of Ad-hVEGF165 on the endothelial cells dysfunction induced by homocysteine (Hcy) and related molecular mechanisms.
METHODSHuman umbilical vein endothelial cells CRL-1730 were treated with Hcy at different concentrations (0, 0.05, 1.00 mmol/L) for 24 h. The same concentration of Hcy, Ad-Track and Ad-hVEGF165 were added to the cells in the following groups: blank group, Hcy0.05 group, Hcy1.00 group, Ad-Track group, Hcy0.05+Ad-Track group, Hcy1.00+Ad-Track group, Ad-hVEGF165 group, Hcy0.05+Ad-hVEGF165 group, Hcy1.00+ Ad-hVEGF165 group for 48 h. The mRNA and protein expressions of eNOS and DDAH2 were detected by real-time PCR and Western blot. The correlations of mRNA and protein expressions between endothelial nitric oxide synthase (eNOS) and dimethylarginine dimthylaminohydrolase (DDAH)2 were evaluated by Pearson correlation analysis.
RESULTSCompared with blank group and Ad-hVEGF165 group, the mRNA and protein expressions of eNOS were decreased in Hcy0.05 group and Hcy0.05+Ad-hVEGF165 group (both P < 0.05), and the mRNA and protein expressions of DDAH2 in cells treated with 0.05 mmol/L and 1.00 mmol/L Hcy were reduced as well (all P < 0.05). DDAH2 mRNA and protein expression are increased (all P < 0.05) in Ad-hVEGF165 group compared with the blank group and Ad-Track, Hcy0.05 + Ad-hVEGF165 and Hcy0.05 group compared with Hcy0.05+Ad-Track group, Hcy1.00+Ad-hVEGF165 and Hcy1.00 group compared with Hcy1.00+Ad-Track group. The mRNA and protein expressions of eNOS and DDAH2 were uncorrelated under the effect of Hcy (r = 0.057 and 0.449, both P > 0.05) and VEGF (r = 0.284 and 0.432, both P > 0.05).
CONCLUSIONRecombinant adenovirus Ad-hVEGF165 could reverse Hcy-induced endothelial cells dysfunction via upregulating the expressions of eNOS and DDAH2.
