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Front. Pharmacol. | doi: 10.3389/fphar.2019.01033

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

Uma M. Sundar1,  Azizah Ugusman2*, Chua K. Hui2,  Jalifah Latip3 and Amilia Aminuddin2
  • 1Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Malaysia
  • 2Department of Physiology, Faculty of Medicine, National University of Malaysia, Malaysia
  • 3Department of Pharmaceutical Chemistry, School of Chemical Sciences & Food Technology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Malaysia

Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of endothelial nitric oxide synthase (eNOS). ADMA is degraded by dimethylarginine dimethylaminohydrolase (DDAH). Elevated level of ADMA leads to reduction in nitric oxide (NO) production that 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 hours. After treatments, HUVEC were collected to measure DDAH 1 mRNA expression using quantitative real time polymerase chain reaction. DDAH 1 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 DDAH 1 mRNA expression (P < 0.05), DDAH 1 protein level (P < 0.01) and DDAH activity (P < 0.05). Treatment with AEPS successfully increased DDAH 1 mRNA expression (P < 0.05), DDAH 1 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.

Keywords: Piper sarmentosum, Asymmetric dimethylarginine, Dimethylarginine dimethylaminohydrolase, tumor necrosis factor-α, endothelial dysfunction, Human umbilical vein endothelial cell (HUVEC)

Received: 28 May 2019; Accepted: 14 Aug 2019.

Edited by:

Steyner F. Cortes, Federal University of Minas Gerais, Brazil

Reviewed by:

Roger L. Santos, Federal University of Espirito Santo, Brazil
Juliana M. Raimundo, Federal University of Rio de Janeiro, Campus Macaé, Brazil  

Copyright: © 2019 Sundar, Ugusman, Hui, Latip and Aminuddin. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Dr. Azizah Ugusman, Department of Physiology, Faculty of Medicine, National University of Malaysia, Cheras, Malaysia,