Effect of the pass of apelin-13 through ADMA-damaged endothelial monolayer on vascular smooth muscle cells
10.3760/cma.j.issn.1001-7097.2011.12.009
- VernacularTitle:apelin-13穿越非对称性二甲基精氨酸损伤的血管内皮层对血管平滑肌细胞的作用
- Author:
Liyan WANG
;
Dongliang ZHANG
;
Yu ZHANG
;
Wenhu LIU
- Publication Type:Journal Article
- Keywords:
Uremia;
Hypertension;
Asymmetric diethylarginine;
Apelin
- From:
Chinese Journal of Nephrology
2011;27(12):907-911
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the effect of apelin-13 on myosin light chain (MLC)phosphorylation in vascular smooth muscle cells under the condition that the permeability of endothelial layer is changed by ADMA.Methods Human umbilical vein endothelial cell (HUVECs) were cultured in bottom of upper well of Transwell system to establish the endothelial monolayer barrier.The cells were cultured normally or incubated with ADMA.Some FITC-apelin-13was added to the upper well.After a period of leakage time,the amount of FITC-apelin-13 in the lower well was measured to evaluate Pa value,which reflected the permeability of endothelial monolayer. Thechangesofcytoskeletonandintercellularjunctionwereobservedby immunofluorescence.Subsequently,Transwell dishes were used to establish a co-culture system with HUVECs in the upper well and human umbilical vein smooth muscle cells(HUVSMCs) in the lower well.There were four groups: blank control group; ADMA group,stimulated by ADMA only;apelin group,apelin-13 used only; ADMA+apelin group,apelin-13 used after HUVECs stimulated by ADMA.Immunofluorescence and Western blotting were used to detect phosphorylation of MLC in HUVSMCs.Results ADMA induced changes of cytoskeleton and intercellular junction and increased endothelial permeability of apelin-13.The percentage of Pa between ADMA-stimulated and control groups varied during the period of apelin-13 pass and peaked at 20 minutes,which was significantly higher than that at 0 minutes [(176.3±9.2)％ vs (100.3±1.5)％,P＜0.05].In the co-culture system,Western blotting showed that p-MLC expression in ADMA +apelin group was significantly higher than that in apelin group(P＜0.05).In ADMA group,a few p-MLC expression was found also.Phosphorylation of MLC in above 3 groups was significantly higher as compared to blank control group(P＜0.05).Conclusions ADMA can increase endothelial permeability of apelin-13 by inducing changes of cytoskeleton and intercellular junction.Apelin-13 passes through the damaged endothelial barrier and directly acts on vascular smooth muscle cells to elevate phosphorylation of MLC,which may play an important role in hypertension of uremic patients.
