[The action of PI3K/AKT during genistein promoting the activity of eNOS].
OBJECTIVE
Genistein could inhibit the development of atherosclerosis. This study explored the role of PI3K/AKT signaling during genistein promoted eNOS activation.


METHODS
Human umbilical vein endothelial cells (HUVECs) were incubated with ox-LDL (100 mg/L), then treated with genistein (100 nmol/L) for 5, 10, 15, 30 and 60 min. The production of NO was assessed by Griess reaction in cell culture supernatant. The mRNA expression of endothelial nitric oxide synthase (eNOS) was detected by reverse transcription-polymerase chain reaction (RT-PCR). The protein expression of eNOS and phosphorylation eNOS(Ser(1179)) were determined by Western blot. The effect of genistein on phosphorylation eNOS(Ser(1179)) level was also observed in the presence of LY294002 or NSC154020 (PI3K and AKT inhibitors).


RESULTS
The concentration of NO and the expression level of phosphorylation eNOS(Ser(1179)) were significantly increased in ox-LDL + genistein treated cells than ox-LDL treated cells (all P < 0.05), and the peak effects were observed at 15 min, however, eNOS mRNA and non-phosphorylated eNOS protein expression were similar between the two groups (P > 0.05). Furthermore, the expression level of phosphorylation eNOS(Ser(1179)) was significantly lower in PIK3/AKT inhibitors LY294002 and NSC154020 treated cells compared with ox-LDL + genistein treated cells (all P < 0.05).


CONCLUSIONS
Genistein could promote the activity of eNOS through increasing phosphorylation eNOS(Ser(1179)) level through PI3K/AKT pathway.
Shanxi Medical University, Taiyuan, China.
Feng-li
FL
Zheng
Jia-hui
JH
Zhao
Hua-ping
HP
Zhang
D002478:Cells, Cultured; D019833:Genistein / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation; D048789:Phytoestrogens / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*
D048789:Phytoestrogens; D019833:Genistein; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt