Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon 200-701, Korea.
J Ginseng Res. 2013 Oct;37(4):413-24. doi: 10.5142/jgr.2013.37.413.
Korean Red Ginseng extract (KRGE) is a traditional herbal medicine utilized to prevent endothelium dysfunction in the cardiovascular system; however, its underlying mechanism has not been clearly elucidated. We here examined the pharmacological effect and molecular mechanism of KRGE on apoptosis of human umbilical vein endothelial cells (HUVECs) in a serum-deprived apoptosis model. KRGE protected HUVECs from serum-deprived apoptosis by inhibiting mitochondrial cytochrome c release and caspase-9/-3 activation. This protective effect was significantly higher than that of American ginseng extract. KRGE treatment increased antiapoptotic Bcl-2 and Bcl-XL protein expression and Akt-dependent Bad phosphorylation. Moreover, KRGE prevented serum deprivation-induced subcellular redistribution of these proteins between the mitochondrion and the cytosol, resulting in suppression of mitochondrial cytochrome c release. In addition, KRGE increased nitric oxide (NO) production via Akt-dependent activation of endothelial NO synthase (eNOS), as well as inhibited caspase-9/-3 activities. These increases were reversed by co-treatment of cells with inhibitors of eNOS and phosphoinositide 3-kinase (PI3K) and pre-incubation of cell lysates in dithiothreitol, indicating KRGE induces NO-mediated caspase modification. Indeed, KRGE inhibited caspase-3 activity via S-nitrosylation. These findings suggest that KRGE prevents serum deprivation-induced HUVEC apoptosis via increased Bcl-2 and Bcl-XL protein expression, PI3K/Akt-dependent Bad phosphorylation, and eNOS/NO-mediated S-nitrosylation of caspases. The cytoprotective property of KRGE may be valuable for developing new pharmaceutical means that limit endothelial cell death induced during the pathogenesis of vascular diseases.
高丽参提取物（KRGE）是一种传统草药，用于预防心血管系统内皮功能障碍；然而，其潜在机制尚未明确阐明。我们在此研究了 KRGE 在血清剥夺凋亡模型中对人脐静脉内皮细胞（HUVEC）凋亡的药理作用和分子机制。KRGE 通过抑制线粒体细胞色素 c 释放和半胱天冬酶-9/-3 激活来保护 HUVEC 免受血清剥夺诱导的凋亡。这种保护作用明显高于西洋参提取物。KRGE 处理增加了抗凋亡 Bcl-2 和 Bcl-XL 蛋白表达以及 Akt 依赖性 Bad 磷酸化。此外，KRGE 防止了这些蛋白在血清剥夺诱导的亚细胞重分布，从而抑制了线粒体细胞色素 c 的释放。此外，KRGE 通过 Akt 依赖性内皮型一氧化氮合酶（eNOS）激活增加了一氧化氮（NO）的产生，同时抑制了半胱天冬酶-9/-3 的活性。用 eNOS 和磷酸肌醇 3-激酶（PI3K）抑制剂共同处理细胞以及用二硫苏糖醇预孵育细胞裂解物，可逆转这些增加，表明 KRGE 通过诱导 NO 介导的半胱氨酸蛋白酶修饰来诱导细胞凋亡。事实上，KRGE 通过 S-亚硝基化抑制了半胱天冬酶-3 的活性。这些发现表明，KRGE 通过增加 Bcl-2 和 Bcl-XL 蛋白表达、PI3K/Akt 依赖性 Bad 磷酸化以及 eNOS/NO 介导的半胱氨酸蛋白酶 S-亚硝基化来防止血清剥夺诱导的 HUVEC 凋亡。KRGE 的细胞保护特性可能对于开发新的药物手段限制血管疾病发病过程中内皮细胞死亡具有重要意义。
