title: PI3K/AKT signaling pathway plays a role in enhancement of eNOS activity by recombinant human angiotensin converting enzyme 2 in human umbilical vein endothelial cells.
abstract: The aim of this study was to investigate the effect of PI3K/AKT signaling pathway in the activity of recombinant human angiotensin converting enzyme 2 (rhACE2) promoted the activity of endothelial nitric oxide synthase (eNOS). The human umbilical vein endothelial cells (HUVEC) were cultured in vitro. Then treated with Ang II (1×10(-6) mol/L) for 24 h. The rhACE2 (100 μmol/L) was added and incubated for 5, 10, 15, 30, 60 min respectively which was based on Ang II intervention. The effect of rhACE2 on phosphorylation eNOS level was also observed in the presence of LY294002 (10 μmol/L) (PI3K/AKT inhibitors). Griess reagent method was applied to measure NO contents in cell culture supernatant, RT-PCR to detect the expression of eNOSmRNA in HUVEC, and Western blot to detect the expression of eNOS and phosphorylated eNOS. In Ang II intervention group, NO contents were significantly lower than control group (P < 0.05). Through rhACE2 treatment, the NO contents in cell culture medium and the expression level of phosphorylated eNOS were significantly higher than in Ang II intervention group (P < 0.05), but eNOSmRNA and non-phosphorylated eNOS protein expression level showed no significant difference (P > 0.05). After HUVEC was intervened by PI3K/AKT pathway inhibitor LY294002, the expression level of phosphorylated eNOS was significantly lower than that in the rhACE2 30 min treatment group (P < 0.05). rhACE2 may reduce the activity of Ang II inhibited endothelial cell eNOS, which can be blocked by PI3K/AKT pathway inhibitor LY294002, suggesting PI3K/AKT signaling pathway plays an important role in rhACE2's promotion of the activity of endothelial cell eNOS.
authors.affiliation: Department of Cardiology, The Second Hospital of Xi'an Jiaotong University Xi'an 710004, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Physiology and Pathophysiology, Medical School of Xi'an Jiaotong University Xi'an 710061, China.
authors.forename: Shi-Jie
authors.initials: SJ
authors.lastname: Wang
authors.affiliation: Department of Cardiology, The Second Hospital of Xi'an Jiaotong University Xi'an 710004, China.
authors.forename: Zhen-Hua
authors.initials: ZH
authors.lastname: Han
authors.affiliation: Department of Cardiology, The Second Hospital of Xi'an Jiaotong University Xi'an 710004, China.
authors.forename: Yong-Qin
authors.initials: YQ
authors.lastname: Li
authors.affiliation: Department of Cardiology, The Second Hospital of Xi'an Jiaotong University Xi'an 710004, China.
authors.forename: Jia-Hong
authors.initials: JH
authors.lastname: Xue
authors.affiliation: Department of Cardiology, The Second Hospital of Xi'an Jiaotong University Xi'an 710004, China.
authors.forename: Deng-Feng
authors.initials: DF
authors.lastname: Gao
authors.affiliation: Department of Cardiology, The Second Hospital of Xi'an Jiaotong University Xi'an 710004, China.
authors.forename: Xiao-San
authors.initials: XS
authors.lastname: Wu
authors.affiliation: Department of Cardiology, The Second Hospital of Xi'an Jiaotong University Xi'an 710004, China.
authors.forename: Cong-Xia
authors.initials: CX
authors.lastname: Wang
mesh_terms: D000804:Angiotensin II / Q000494:pharmacology*; D000085962:Angiotensin-Converting Enzyme 2; D004791:Enzyme Inhibitors / Q000494:pharmacology; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D007703:Peptidyl-Dipeptidase A / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010766:Phosphorylation / Q000187:drug effects; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology
chemical_list: D000804:Angiotensin II; D000085962:Angiotensin-Converting Enzyme 2; D004791:Enzyme Inhibitors; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III; D007703:Peptidyl-Dipeptidase A; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt; C496314:NOS3 protein, human; C000705307:ACE2 protein, human
keywords: PI3K/AKT signaling pathway; Recombinant human ACE2; nitric oxide synthase