The effects of ginsenoside Rb1 on endothelial damage and ghrelin expression induced by hyperhomocysteine

ObjectiveStudies have indicated that ginsenoside Rb1 and ghrelin could both prevent homocysteine (Hcy)-induced endothelial dysfunction through the endothelial nitric oxide synthase (eNOS)/nitric oxide (NO) mechanism. This study investigated whether endogenous ghrelin mediates the endothelial protection of ginsenosidee Rb1 through in vitro and in vivo experiments.MethodsRats were randomized into a control group, a hyperhomocysteine (HHcy) model group with a high methionine diet, a ginsenosides (GS) group, and HHcy plus GS group. Plasma ghrelin was detected by enzyme-linked immunosorbent assay. Aortic rings for control and HHcy groups were treated with ghrelin or not. Endothelium-dependent vasodilatation function was evaluated by the aortic ring assay, and the structural changes were visualized by hematoxylin and eosin staining. Human umbilical vein endothelial cells (HUVECs) were cultured, and the experimental conditions were optimized according to NO production. After treatment, the NO, ghrelin, and von Willebrand factor (vWF) levels in the media were detected and analyzed with linear regression. Ghrelin and eNOS expression were observed by cell immunohistochemical staining. Ghrelin receptor antagonist was used to detect the mechanism of ginsenoside Rb1 on NO production, which was reflected by diacetylated 4,5-diaminofluorescein-2 diacetate fluorescence.ResultsIn vivo experiments demonstrated that plasma ghrelin levels in the HHcy group were significantly elevated vs controls (P < .05) and were significantly increased in the HHcy plus GS group (P < .01). Compared with control, endothelium-dependent vasodilatation function was greatly reduced in the HHcy group (P < .01), which was significantly increased in HHcy plus ghrelin group compared with HHcy group (P < .01). The arterial walls of HHcy group exhibited characteristic pathologic changes, which were repaired in HHcy plus ghrelin group. In vivo, compared with Hcy (200 μM) group, HUVECs pretreated with ginsenoside Rb1 (10 μM) for 30 minutes showed significant increases in NO and ghrelin levels and evident reduction in vWF levels. Linear regression analysis demonstrated that ghrelin levels were significantly positively correlated with NO levels and significantly negatively correlated with vWF levels. The addition of Rb1 to Hcy also greatly reversed Hcy-induced downregulation of ghrelin and eNOS expression. Ghrelin inhibition significantly abolished the upregulation of NO levels induced by Rb1.ConclusionGhrelin can prevent Hcy-induced vascular endothelial dysfunction and structural damage. The compensatory elevation of plasma ghrelin levels in an Hcy-induced endothelial injury model may be a protective response. Ginsenoside Rb1 can significantly stimulate the ghrelin endocrine to inhibit endothelial injury. Ginsenoside also upregulates the NO signaling pathway reduced by Hcy through the ghrelin molecular mechanism.

Clinical RelevanceHomocysteine is an independent risk factor for endothelial injury and dysfunction. Ginsenoside Rb1, a major constituent of ginseng (traditional Chinese herb), can effectively block homocysteine-induced dysfunction of endothelium-dependent vasorelaxation as well as endothelial nitric oxide synthase (eNOS) downregulation. Ghrelin, a novel brain-gut peptide, has multiple cardiovascular protective effects, including vasorelaxation. The objective of this study was to determine the effect and molecular changes of ginsenoside Rb1. Our results showed that ginseng compounds have effects of vasorelaxation and endogenous ghrelin upregulation. Exogenous ghrelin can block homocysteine-induced endothelial injury and dysfunction in the rat thoracic aortic. Ginsenoside Rb1 can improve NO production, ghrelin, and eNOS expression in human umbilical vascular endothelial cells. This study indicated that ginsenoside Rb1 has a prospective clinical future in controlling homocysteine-associated vascular diseases through the ghrelin molecule and NO-signaling mechanism.