Korean J Physiol Pharmacol.  2014 Apr;18(2):95-101. 10.4196/kjpp.2014.18.2.95.

Korean Red Ginseng Water Extract Restores Impaired Endothelial Function by Inhibiting Arginase Activity in Aged Mice

Affiliations
  • 1Department of Biology, Kangwon National University, Chuncheon 200-701, Korea. ryoosw08@kangwon.ac.kr
  • 2Department of Anesthesiology and Pain Medicine, Yonsei University Wonju College of Medicine, Wonju 220-701, Korea. hyunkyolim@yonsei.ac.kr

Abstract

Cardiovascular disease is the prime cause of morbidity and mortality and the population ages that may contribute to increase in the occurrence of cardiovascular disease. Arginase upregulation is associated with impaired endothelial function in aged vascular system and thus may contribute to cardiovascular disease. According to recent research, Korean Red Ginseng water extract (KRGE) may reduce cardiovascular disease risk by improving vascular system health. The purpose of this study was to examine mechanisms contributing to age-related vascular endothelial dysfunction and to determine whether KRGE improves these functions in aged mice. Young (10+/-3 weeks) and aged (55+/-5 weeks) male mice (C57BL/6J) were orally administered 0, 10, or 20 mg/mouse/day of KRGE for 4 weeks. Animals were sacrificed and the aortas were removed. Endothelial arginase activity, nitric oxide (NO) generation and reactive oxygen species (ROS) production, endothelial nitric oxide synthase (eNOS) coupling, vascular tension, and plasma peroxynitrite production were measured. KRGE attenuated arginase activity, restored nitric oxide (NO) generation, reduced ROS production, and enhanced eNOS coupling in aged mice. KRGE also improved vascular tension in aged vessels, as indicated by increased acetylcholine-induced vasorelaxation and improved phenylephrine-stimulated vasoconstriction. Furthermore, KRGE prevented plasma peroxynitrite formation in aged mice, indicating reduced lipid peroxidation. These results suggest KRGE exerts vasoprotective effects by inhibiting arginase activity and augmenting NO signaling and may be a useful treatment for age-dependent vascular diseases.

Keyword

Aging; Endothelial nitric oxide synthase; Korean red ginseng extract; Vasorelaxation

MeSH Terms

Aging
Animals
Aorta
Arginase*
Cardiovascular Diseases
Humans
Lipid Peroxidation
Male
Mice*
Mortality
Nitric Oxide
Nitric Oxide Synthase Type III
Panax*
Peroxynitrous Acid
Plasma
Reactive Oxygen Species
Up-Regulation
Vascular Diseases
Vasoconstriction
Vasodilation
Water*
Arginase
Nitric Oxide
Nitric Oxide Synthase Type III
Peroxynitrous Acid
Reactive Oxygen Species
Water

Figure

  • Fig. 1 HPLC analysis of Korean Red Ginseng Extract.

  • Fig. 2 Increased arginase activity in aged mice aorta was attenuated by Korean Red Ginseng Extract administration. Oral administration of Korean Red Ginseng Extract for 4 weeks resulted in decreased arginase activity in aged mice. * vs. young untreated, p<0.01; ** vs. aged untreated, p<0.01.

  • Fig. 3 Korean Red Ginseng Extract restored impaired endothelial function in aged mice. Isolated aortic segments were incubated with 4,5-Diaminofluorescein diacetate (DAF-AM, 5 µM) and fluorescence was measured in real-time (endothelium side up). The slope of DAF-AM fluorescence was determined. (A) Korean Red Ginseng Extract (KRGE) administration increased the slope of DAF fluorescence in young vessels (# vs. young untreated, p<0.01). The slope of DAF fluorescence was lower in aged mouse aorta (* vs. young untreated, p<0.01) and increased after KRGE ingestion (** vs. aged untreated, p<0.01; n=4 mice). NG-Nitroarginine Methyl Ester (L-NAME) was used as a control. (B) Reactive Oxygen species (ROS) production in aortic endothelium was measured with dihydroethidium (DHE, 5 µM), and the slope of DHE fluorescence was determined using cumulative data. KRGE intake decreased ROS production in both young and old mice (# vs. young untreated, p<0.01; * vs. young untreated, p<0.01; ** vs. aged untreated, p<0.01; ## vs. aged untreated, p<0.01; n=4 mice). MnTBAP was used as a control.

  • Fig. 4 Korean Red Ginseng Extract improves age-dependent endothelial Nitric Oxide Synthase uncoupling. (A) KRGE did not affect protein expression in the aortas of young or aged mice. (B) Endothelial nitric oxide synthase (eNOS) dimerization was analyzed by low-temperature sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. The eNOS was more uncoupled in aged mouse aorta (* vs. young untreated, p<0.05). (C) Korean Red Ginseng Extract (KRGE) increased eNOS coupling in aged mouse aorta (* vs. aged untreated, p<0.05).

  • Fig. 5 Korean Red Ginseng Extract improved vascular reactivity impairments in aged mice. (A) Endothelium-dependent relaxation response to acetylcholine (Ach) was impaired in aged aortas (*young vs. aged, p<0.01). Korean Red Ginseng Extract (KRGE) administration improved relaxation (**aged untreated vs. aged+KRGE, p<0.05). (B) Aged aortic vessels had attenuated contractile responses to phenylephrine (PE) compared to young mice (*young vs. aged, p<0.01). KGRE restored the PE-mediated pressor response in aged aortic vessels (**aged untreated vs. aged+KRGE, p<0.01) and attenuated the responses in young aortic vessels (# young vs. young+KRGE, p<0.05).

  • Fig. 6 Korean Red Ginseng Extract administration reduced peroxynitrite formation. Peroxynitrite content in plasma was measured by Thiobarbituric acid reactive substances (TBARS) assay. Aged mice had greater peroxynitrite content than their young counterparts (*young vs. aged, p<0.01; n=6). Korean Red Ginseng Extract (KRGE) reduced peroxynitrite content in aged mice (# aged untreated vs. aged+KRGE, p<0.01; n=6).


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