[Effect of puerarin on ADMA-DDAH system in human umbilical vein endothelial cells cultured with oxidized free radical].
OBJECTIVE
To observe the effects of puerarin on activity of dimethylarginine dimethylaminohydrolase (DDAH) in human umbilical vein endothelial cells (HUVECs) cultured with oxidized free radical (OFR), to explore the effect of puerarin on metabolic mechanism of asymmetric dimethylarginine (ADMA).


METHODS
HUVECs of the 3rd - 6th passage cultured with modified Jaffe's method were divided into 4 groups, the blank control group cultured with DMEM medium, the OFR group cultured with DMEM medium containing 0.1 mmol of OFR per liter, the puerarin group 1 and 2 cultured with DMEM medium containing 0.1 mmol of OFR per liter as well as 0.5 mg/ml and 1.0 mg/ml of puerarin respectively. After being incubated for 24 h, activity of nitric oxide synthase (NOS), contents of nitric oxide (NO), ADMA, endothelin (ET), and L-citrulline (L-cit) in the supernate were measured, and DDAH protein expression in the lysate was detected by Western blotting.


RESULTS
Compared with those in the blank control group, ADMA and ET contents were higher, while the levels of NO and L-cit and the activity of NOS were lower markedly, but the DDAH expression changed insignificantly in the OFR group. These abnormalities were restored significantly in the puerarin groups.


CONCLUSIONS
The increase of ADMA in OFR injured HUVECs was correlated with the reduction of DDAH activity and irrelevant to DDAH expression. Puerarin could promote ADMA metabolism through increasing DDAH activity, and improve NOS activity, thus to reduce the impairing of OFR on endothelial function.
Li
Ju-Xiang
JX
Department of Cardiology, The Second Affiliated Hospital of Jiangxi Medical College, Nanchang. ysj040101@yahoo.com.cn
Chen
Jing
J
Deng
Zhi-Hua
ZH
D000581:Amidohydrolases / Q000378:metabolism*; D001120:Arginine / Q000031:analogs & derivatives* / Q000378:metabolism; D015153:Blotting, Western; D002478:Cells, Cultured; D003470:Culture Media; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D016232:Endothelins / Q000378:metabolism; D005609:Free Radicals / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D007529:Isoflavones / Q000494:pharmacology*; D009566:Nitrates / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000378:metabolism; D009573:Nitrites / Q000378:metabolism; D010084:Oxidation-Reduction / Q000187:drug effects; D014471:Umbilical Veins / Q000166:cytology
D003470:Culture Media; D016232:Endothelins; D005609:Free Radicals; D007529:Isoflavones; D009566:Nitrates; D009573:Nitrites; D009569:Nitric Oxide; C018524:N,N-dimethylarginine; D001120:Arginine; D019001:Nitric Oxide Synthase; D000581:Amidohydrolases; C059999:dimethylargininase; C033607:puerarin