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Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of endothelial nitric oxide synthase (eNOS). ADMA is degraded by dimethylarginine dimethylaminohydrolase (DDAH). Elevated levels of ADMA lead to reduction in nitric oxide (NO) production, which is linked to endothelial dysfunction and atherosclerosis.  Piper sarmentosum  is an herb that has shown stimulation on endothelial NO production by increasing both expression and activity of eNOS. Thus, this study determined whether the positive effect of  P. sarmentosum  on NO production is related to its modulation on the DDAH–ADMA pathway in cultured human umbilical vein endothelial cells (HUVEC) exposed to tumor necrosis factor-α (TNF-α). HUVEC were divided into four groups: control, treatment with 250 µg/ml of aqueous extract of  P. sarmentosum  leaves (AEPS), treatment with 30 ng/ml of TNF-α, and concomitant treatment with AEPS and TNF-α for 24 h. After treatments, HUVEC were collected to measure  DDAH1  messenger RNA (mRNA) expression using quantitative real-time polymerase chain reaction. DDAH1 protein level was measured using enzyme-linked immunosorbent assay (ELISA), and DDAH enzyme activity was measured using colorimetric assay. ADMA concentration was measured using ELISA, and NO level was measured using Griess assay. Compared to control, TNF-α-treated HUVEC showed reduction in  DDAH1  mRNA expression ( P  < 0.05), DDAH1 protein level ( P  < 0.01), and DDAH activity ( P  < 0.05). Treatment with AEPS successfully increased  DDAH1  mRNA expression ( P  < 0.05), DDAH1 protein level ( P  < 0.01), and DDAH activity ( P  < 0.05) in TNF-α-treated HUVEC. Treatment with TNF-α caused an increase in ADMA level ( P  < 0.01) and a decrease in endothelial NO production ( P  < 0.001). Whereas treatment with AEPS was able to reduce ADMA level ( P  < 0.01) and restore NO ( P  < 0.001) in TNF-α-treated HUVEC. The results suggested that AEPS promotes endothelial NO production by stimulating DDAH activity and thus reducing ADMA level in TNF-α-treated HUVEC.

frontiers in pharmacology

Piper sarmentosum Promotes Endothelial Nitric Oxide Production by Reducing Asymmetric Dimethylarginine in Tumor Necrosis Factor-α-Induced Human Umbilical Vein Endothelial Cells