摘要
目的研究人参皂苷Rg1对S-亚硝基化谷胱甘肽(S-nitrosoglutathione,GSNO)引起内皮细胞损伤的保护作用,并初步探讨可能的分子机制。方法建立S-亚硝基化谷胱甘肽损伤内皮细胞模型,四甲基偶氮唑蓝法检测S-亚硝基化谷胱甘肽对内皮细胞存活率影响,4,6-二脒基-2-苯基吲哚(DAPI)染色观察细胞核聚缩。人参皂苷Rg1 10,20,40μmol·L-1预处理内皮细胞24 h,加S-亚硝基化谷胱甘肽损伤观察细胞存活率。乳酸脱氢酶试剂盒检测乳酸脱氢酶释放。JC-1染色检测线粒体膜电位(mitochondrial membrane potential,ΔΨm)变化。Western blot法检测bcl-2,bax,cytochrome C(cyt C),cleaved caspase 9,procaspase 9,cleaved caspase 3,pro-caspase 3蛋白表达。结果 S-亚硝基化谷胱甘肽抑制内皮细胞增殖。Rg1(20,40μmol·L-1)预处理24 h可显著减弱S-亚硝基化谷胱甘肽引起内皮细胞存活率降低(P<0.05)和乳酸脱氢酶释放(P<0.05)。Rg1可抑制S-亚硝基化谷胱甘肽引起的ΔΨm下降,抑制S-亚硝基化谷胱甘肽引起的线粒体依赖凋亡蛋白bcl-2/bax,cyt C,cleaved caspase 9/caspase 9,cleaved caspase 3/caspase 3蛋白表达上调,提示Rg1可抑制内皮细胞线粒体依赖凋亡。结论人参皂苷Rg1可减少S-亚硝基化谷胱甘肽引起内皮细胞凋亡,其作用机制与抑制线粒体依赖凋亡通路密切相关。
OBJECTIVE To investigate the role of ginsenoside Rgl effect on S-nitrosoglutathione （GSNO） induced human endothelial cells injury, and to further find the potential mechanisms of Rgl effects. METHODS The cell viability was measured by MTT assay. Cell viability and LDH release were observed after pre-treatment of Rgl （ 10, 20 and 40 μmol·L^-1 ） for 24 h. JC-1 staining was used to detecte endothelial cells mitochondrial membrane potential （△ψm）. Bcl-2, bax, cytochrome C （cyt C）, pro-caspase 9, cleaved caspase 9, pro-caspase 3 and cleaved caspase 3 were detected by Western blot assay. RESULTS GSNO caused cell toxicity in a time- and dosedependent manner. Rgl protected endothelial cells against GSNO-induced injury and LDH release. Rgl reduced GSNO-induced AWm decrease. GSNO induced upregulation of cyt C release, cleaved caspase 9 and cleaved caspase 3 in the cytoplasm and decreased the ratio of bcl-2/bax in mitochondrial. This data suggested that Rgl protected endothelial cells against GSNO-induced injury via inhibiting mitochondrial dependent apoptosis signaling pathway. CONCLUSION Ginsenoside Rgl attunate GSNO-induced endothelial cells injury via inhibiting mitochondrial apoptotic cascades.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2014年第14期1211-1215,共5页
Chinese Pharmaceutical Journal
基金
浙江省自然科学基金资助项目(Y2100564)
浙江省中医药管理局基金资助项目(2010ZA004
2007CA097)
作者简介
严洁萍，女，博士，药师研究方向：心脑血管药理
通讯作者：吕良忠，男，博士，主任药师，副教授，硕士生导师研究方向：心脑血管及肿瘤药理研究Tel／Fax：（0571）85893122E-mail：Miangzhong@126．com
