Inhibitory Effect of Ginsenoside Rg1 on Vascular Smooth Muscle Cell Proliferation Induced by PDGF-BB Is Involved in Nitric Oxide Formation

Jing Huang,Li-Sheng Li,Dan-Li Yang,Xie-Nan Huang,Jiang Deng,Qi-Hai Gong

doi:10.1155/2012/314395

Abstract

Ginsenoside Rg1 (Rg1) has been reported to suppress the proliferation of vascular smooth muscle cells (VSMCs). This study aimed to observe the role of nitric oxide (NO) in Rg1-antiproliferative effect. VSMCs from the thoracic aorta of SD rats were cultured by tissue explant method, and the effect of Rg1 (20 mg·L−1, 60 mg·L−1, and 180 mg·L−1) on platelet-derived growth factor-BB (PDGF-BB)-induced proliferation was evaluated by MTT assay. The cell cycle was analyzed by flow cytometry. For probing the mechanisms, the content of NO in supernatant and cGMP level in VSMCs was measured by nitric oxide kit and cGMP radio-immunity kit, respectively; the expressions of protooncogene c-fos and endothelial NO synthase (eNOS) mRNA in the VSMCs were detected by real-time RT-PCR; the intracellular free calcium concentration ([Ca2+]i) was detected with Fura-2/AM-loaded VSMCs. Comparing with that in normal group, Rg1 180 mg·L−1 did not change the absorbance of MTT and cell percent of G0/G1, G2/M, and S phase in normal cells (P > 0.05). Contrarily, PDGF-BB could increase the absorbance of MTT (P < 0.01) and the percent of the S phase cells but decrease the G0/G1 phase cell percent in the cell cycle, accompanied with an upregulating c-fos mRNA expression (P < 0.01), which was reversed by additions of Rg1(20 mg·L−1, 60 mg·L−1, and 180 mg·L−1). Rg1 administration could also significantly increase the NO content in supernatant and the cGMP level in VSMCs, as well as the eNOS mRNA expression in the cells, in comparison of that in the group treated with PDGF-BB alone (P < 0.01). Furthermore, Rg1 caused a further increase in the elevated [Ca2+]i induced by PDGF-BB. It was concluded that Rg1 could inhibit the VSMC proliferation induced by PDGF-BB through restricting the G0/G1 phase to S-phase progression in cell cycle. The mechanisms may be related to the upregulation of eNOS mRNA and the increase of the formation of NO and cGMP.

Highlights

Ginsenoside Rg1 (Rg1) is believed to be one of the main active principles in ginseng
In normal cells without any treatment of growth factor, Rg1 180 mg · L−1 did not change the MTT absorbance, while the addition of platelet-derived growth factor-BB (PDGF-BB) could significantly increase the absorbance in MTT assay (P < 0.01), which tended to be inhibited by an addition of Rg1 20 mg · L−1 (P > 0.05), and was remarkably inhibited by Rg1 60 mg · L−1and 180 mg · L−1 (P < 0.05 and P < 0.01), suggesting the antiproliferating effect of Rg1 on vascular smooth muscle cells (VSMCs) (Figure 1)
VSMC proliferation has been known to be an important component of vessel wall remodelling in response to injury, such as after angioplasty and during atherosclerosis formation [23, 24]

Summary

Introduction

Ginsenoside Rg1 (Rg1) is believed to be one of the main active principles in ginseng A. Meyer), a traditional Chinese medicine used to enhance stamina and capacity to cope with fatigue as well as physical stress. It has been reported that Rg1 has many beneficial effects on several systems. Rg1 has been reported to inhibit proliferation of vascular smooth muscle cells (VSMCs) induced by tumor necrosis factor-α and block the cell cycle in the G1-phase via depressing the signaling pathways of ERK, PI3K/PKB, and PKC [4, 5]. Cell proliferation is modulated by many factors, more studies should be done to elucidate the action mechanisms of the antiproliferative effect of Rg1

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