Wei Zhou, MD, Hong Chai, MD, PhD, Peter H. Lin, MD, Alan B

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Ginsenoside Rb1 blocks homocysteine-induced endothelial dysfunction in porcine coronary arteries Wei Zhou, MD, Hong Chai, MD, PhD, Peter H. Lin, MD, Alan B. Lumsden, MD, Qizhi Yao, MD, PhD, Changyi Chen, MD, PhD Journal of Vascular Surgery Volume 41, Issue 5, Pages 861-868 (May 2005) DOI: 10.1016/j.jvs.2005.01.054 Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions

Fig 1 The effects of Hcy and ginsenoside Rb1 on the contractility of porcine coronary arteries. The rings (n = 8 per group) were cultured in DMSO control, Hcy (50 μM) alone, low-dose Rb1 (1 μM) or high-dose Rb1 (10 μM) alone, and Hcy combined with low-dose or high-dose Rb1. There is no substantial difference in maximum vessel contraction in response to U46619 (10−7 M) among all groups (n = 8, P > .05). Journal of Vascular Surgery 2005 41, 861-868DOI: (10.1016/j.jvs.2005.01.054) Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions

Fig 2 The effects of Hcy and GS Rb1 on the endothelium-dependent vasorelaxation induced by bradykinin. A, Vasorelaxation in response to cumulative doses of bradykinin showing improved vasorelaxation among all groups as the concentration of bradykinin increases. The high-dose Rb1-Hcy group showed significant improvement in vasorelaxation compared with Hcy alone at the bradykinin concentrations of 10−7, 10−6, and 10−5 M. B, Vasorelaxation in response to the highest dose of bradykinin (10−5 M). The Hcy and low-dose Rb1-Hcy groups showed significant impairments in vasorelaxation compared with the control (n = 8, P < .05), whereas the high-dose Rb1-Hcy group showed significant improvement in vasorelaxation compared with Hcy (n = 8, P < .01) and similar to the control group. Journal of Vascular Surgery 2005 41, 861-868DOI: (10.1016/j.jvs.2005.01.054) Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions

Fig 3 The effects of Hcy and GS Rb1 on endothelium-independent relaxation as compared with the DMSO control. There is no significant difference among all groups (n = 8, P > .05) in the endothelium-independent vasorelaxation induced by SNP. Journal of Vascular Surgery 2005 41, 861-868DOI: (10.1016/j.jvs.2005.01.054) Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions

Fig 4 The influence of Hcy and GS Rb1 on mRNA expression in the treated arterial rings. The levels of eNOS mRNA expression in porcine coronary arteries were measured using real-time PCR analysis. The relative expression for the eNOS in each sample was normalized against GAPDH, shown as 2^[Ct(GAPDH)-Ct(eNOS)]. The Hcy-alone group showed a significant reduction in eNOS mRNA level compared with control (n = 6, P < .05), and Rb1 (high-dose) significantly improved eNOS mRNA level in the Rb1-Hcy groups (n = 6, P < .05). Journal of Vascular Surgery 2005 41, 861-868DOI: (10.1016/j.jvs.2005.01.054) Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions

Fig 5 The level of eNOS protein production determined by Western blot analysis after being normalized with β-actin as an internal control. Hcy significantly impaired eNOS protein level compared with the control (P < .01), and the high-dose Rb1-Hcy combined group showed statistically significant improvement in eNOS protein levels as compared with the Hcy-alone group (n = 3, P < .05). Journal of Vascular Surgery 2005 41, 861-868DOI: (10.1016/j.jvs.2005.01.054) Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions

Fig 6 Immunohistochemical staining of eNOS protein expression in porcine coronary arterial endothelium. A, DMSO control group showing a strong positive staining pattern of eNOS on the endothelial layer of the arterial walls. B, Hcy-alone group showing poor staining pattern of eNOS. C, High-dose Rb1-alone group, and D, high-dose Rb1-Hcy combined group showing a strong positive staining pattern similar to that of the control group. Journal of Vascular Surgery 2005 41, 861-868DOI: (10.1016/j.jvs.2005.01.054) Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions

Fig 7 Levels of superoxide anion produced by endothelial cells using the lucigenin-enhanced chemiluminescence. Superoxide anion was markedly increased in Hcy-treated groups compared with DMSO control (n = 6, P < .01). High-dose or low-dose Rb1 alone showed no effects on the level of peak chemiluminescence as compared with the control. Both high-dose and low-dose Rb1-Hcy combined groups showed a substantial reduction in superoxide anion levels. Journal of Vascular Surgery 2005 41, 861-868DOI: (10.1016/j.jvs.2005.01.054) Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions