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羟基红花黄色素A对缺氧条件下人脐静脉内皮细胞的影响

Effect of Hydroxysafflor yellow A on human umbilical vein endothelial cells under hypoxia.

作者信息

Ji Deng Bo, Zhang Li Yun, Li Chang Ling, Ye Jia, Zhu Hai Bo

机构信息

Department of Molecular and Cellular Pharmacology, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China.

出版信息

Vascul Pharmacol. 2009 Mar-Apr;50(3-4):137-45. doi: 10.1016/j.vph.2008.11.009. Epub 2008 Nov 28.

Abstract

Hydroxysafflor yellow A (HSYA), is a component of the flower, Carthamus tinctorius L. In this study, we investigated its effect on Human Umbilical Vein Endothelial Cells (HUVECs) under hypoxia. We evaluated cell viability using the MTT kit. The cell cycle distribution was analyzed by PI staining flow cytometric analysis. PI AnnexinV-FITC detection and the TUNEL assay were performed to evaluate the apoptosis rate. Nitric oxide (NO) generation in cell supernatant was measured by the Griess assay. RT-PCR, Western blot and Immunocytochemistry analysis were used to evaluate the changes of Bcl-2, Bax, p53 and eNOS. Our data showed that HSYA inhibited cell apoptosis and cell cycle G1 arrest induced by hypoxia. HSYA treatment increased the Bcl-2/Bax ratio of protein and mRNA, reduced p53 protein expression in cell nucleus. In addition, HSYA enhanced the NO content of cell supernatant under hypoxia, accompanied with upregulating eNOS mRNA expression and protein level. Taken together, these results demonstrate that HSYA could protect HUVECs from hypoxia induced injuries by inhibiting cell apoptosis and cell cycle arrest. These findings have partly revealed the molecular mechanism of HSYA on treating of ischemic heart disease. We expected our experiments might provide some clues for further research.

摘要

羟基红花黄色素A(HSYA)是红花(Carthamus tinctorius L.)花中的一种成分。在本研究中,我们研究了其在缺氧条件下对人脐静脉内皮细胞(HUVECs)的影响。我们使用MTT试剂盒评估细胞活力。通过PI染色流式细胞术分析细胞周期分布。进行PI AnnexinV-FITC检测和TUNEL检测以评估凋亡率。通过Griess检测法测量细胞上清液中一氧化氮(NO)的生成。使用RT-PCR、蛋白质印迹法和免疫细胞化学分析来评估Bcl-2、Bax、p53和eNOS的变化。我们的数据表明,HSYA抑制缺氧诱导的细胞凋亡和细胞周期G1期阻滞。HSYA处理增加了蛋白质和mRNA的Bcl-2/Bax比值,降低了细胞核中p53蛋白的表达。此外,HSYA在缺氧条件下提高了细胞上清液中的NO含量,同时上调了eNOS mRNA表达和蛋白水平。综上所述,这些结果表明HSYA可以通过抑制细胞凋亡和细胞周期阻滞来保护HUVECs免受缺氧诱导的损伤。这些发现部分揭示了HSYA治疗缺血性心脏病的分子机制。我们期望我们的实验可能为进一步的研究提供一些线索。

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