Research Of The Effect Of Doxazosin On Indoxyl Sulfate-Induced MCP-1 Expression

Objectives:Chronic kidney disease (CKD) is recognized as a common condition that elevates the risk of atherosclerotic cardiovascular disease (CVD). Evidence suggests that accumulated uremic toxin in patients with CKD can induce oxidative stress reaction, which is an emerging key mechanism of atherosclerosis in CKD. This study aimed to determine if doxazosin-an inhibitor of a1-adrenoceptor affects the signaling pathway that is activated by indoxyl sulfate (IS)-a uremic toxin to induce monocyte chemoattractant protein-1 (MCP-1), which plays an important role in the development of atherosclerosis, in cultured human umbilical vein endothelial cells (HUVEC).Methods:In order to achieve these goals, HUVEC were incubated with IS and/or doxazosin. The production of reactive oxygen species (ROS) in HUVEC was assessed by dihydroethidium (DHE) staining. The expression of ERK1/2 phosphorylation,p38 phosphorylation and NF-κB p65 was studied by western blotting. The expression of monocyte chemoattractant protein-1 (MCP-1) in the supernatant was measured using a fluorescence microplate reader (ELISA). The expression of MCP-1 and a1-adrenoceptor D,B,A mRNA were analysed by reverse transcription polymerase chain reaction (RT-PCR).Results:(1) Doxazosin significantly inhibited IS-induced ROS generation in HUVEC.(2) Doxazosin inhibited IS-induced ERK1/2 phosphorylation in HUVEC.(3) Doxazosin showed no inhibitory effect on p38 MAPK phosphorylation in HUVEC.(4) Doxazosin reduced the nuclear translocation of NF-κB p65 in HUVEC.(5) The expression of MCP-1 mRNA which was induced by IS could be inhibited by doxazosin in HUVEC.(6) The expression of MCP-1 protein which was induced by IS could be inhibited by doxazosin in HUVEC. (7) The mRNA of a1-adrenoceptor D and A were detected in HUVEC, but we didn't find a1-adrenoceptor B in HUVEC.Conclusions:Doxazosin reduces IS-induced MCP-1 expression in HUVEC by inhibits ERK1/2/NF-κB pathway; these demonstrate for the first time that doxazosin may have significant anti-oxidative stress and anti-artherosclerosis properties in HUVEC. We confirm the existence of a1-adrenoceptor D and a1-adrenoceptor A in HUVEC, which may help to discover the mechanism underlying the anti-artherosclerosis effects of doxazosin in HUVEC.