Effect of intermittent high glucose on synthesis of nitric oxide in human umbilical vein endothelial cells and its mechanism.
OBJECTIVE
To compare the effect of intermittent and constant high glucose media on the synthesis of vascular relaxing factor nitric oxide (NO) in cultured human umbilical vein endothelial cells (HUVECs), and to investigate its mechanism.


METHODS
HUVECs were exposed to 5.5 mmol/L glucose (normal control), 20 mmol/L glucose (constant high glucose), and 5.5 mmol/L alternating with 20 mmol/L glucose (intermittent high glucose) for 7 days. The NO level was measured using Griess Reaction in cell culture supernatants; the expressions of PI3K, PKB and eNOS mRNA and protein were measured with RT-PCR or Western blot.


RESULTS
NO level in the intermittent high glucose group was significantly lower than that in the constant high glucose group (P<0.05). The expression of PI3K, PKB, and eNOS in the intermittent high glucose group was significantly lower than that in the constant high glucose group (P<0.05).


CONCLUSIONS
Intermittent high glucose could be more deleterious to endothelial cells than constant high glucose, and may lead to decrease synthesis of NO by inhibiting PI3K/PKB/eNOS pathway activation.
Liao
Jie
J
Department of Endocrinology, Xiangya Hospital, Central South University, Changsha 410008, China.
Lei
Minxiang
M
Chen
Xiong
X
Liu
Feng
F
D002478:Cells, Cultured; D003470:Culture Media; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D009569:Nitric Oxide / Q000096:biosynthesis*; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014471:Umbilical Veins / Q000166:cytology*
D003470:Culture Media; D012333:RNA, Messenger; D009569:Nitric Oxide; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D005947:Glucose