Calycosin-7-O-β-D-glucoside promotes oxidative stress-induced cytoskeleton reorganization through integrin-linked kinase signaling pathway in vascular endothelial cells.
Calycosin-7-O-β-D-glucoside promotes oxidative stress-induced cytoskeleton reorganization through integrin-linked kinase signaling pathway in vascular endothelial cells.
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毛蕊花苷-7-O-β-D-葡萄糖苷通过整合素连接激酶信号通路促进血管内皮细胞氧化应激诱导的细胞骨架重组
DOI：
10.1186/s12906-015-0839-5
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发表时间：
2015-09-07
影响因子：
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通讯作者：
Xu JX
中科院分区：
文献类型：
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作者：
Jiang YH;Sun W;Li W;Hu HZ;Zhou L;Jiang HH;Xu JX
血管内皮功能障碍涉及许多病理情况。细胞骨架在血管内皮通透性屏障和炎症反应中起重要作用。许多中药具有内皮保护作用，其中黄芪是治疗心血管疾病和肾脏疾病的名贵中药。本研究旨在探讨黄芪的主要有效单体成分毛蕊异黄酮（calycosin）对细菌内毒素（LPS）诱导的内皮细胞损伤的保护作用。用脂多糖（LPS）诱导人脐静脉内皮细胞（HUVECs）损伤。观察毛蕊异黄酮对LPS诱导的细胞活力、凋亡率、细胞迁移、一氧化氮合酶（NOS）、细胞内活性氧（ROS）产生和细胞骨架结构的影响。采用基因芯片技术筛选可能的基因表达变化。在基因芯片检测结果的基础上，采用实时荧光定量RT-PCR或western blot方法进一步分析Rho/ROCK通路和AKT通路相关基因的表达谱。毛蕊异黄酮可提高细胞活力，抑制细胞凋亡，并保护细胞免受LPS诱导的细胞迁移和ROS产生的减少，NOS的蛋白水平与Y27632和缬沙坦相当。与Y27632和缬沙坦类似，毛蕊异黄酮也中和LPS诱导的肌动球蛋白收缩和黏着斑蛋白聚集。基因芯片、实时荧光定量PCR和western blot结果显示，LPS诱导FN、ITG A5、RhoA、PI 3 K（或western blot中的PIP 2）、FAK、VEGF和VEGF R2的表达，抑制MLCP的表达。我们认为毛蕊异黄酮对HUVECs的调控可能涉及多条途径。毛蕊异黄酮可能通过促进NO的产生，减少PMLC，抑制Rho/ROCK通路激活引起的细胞骨架重构，降低VEGF、VEGF R2和PI 3 K水平，抑制AKT通路，从而激活MLCP。毛蕊异黄酮可能通过抑制Rho/ROCK通路和调节AKT通路保护HUVEC免受LPS诱导的内皮损伤。
Dysfunction of vascular endothelium is implicated in many pathological situations. Cytoskeleton plays an importance role in vascular endothelial permeability barrier and inflammatory response. Many Chinese herbs have the endothelial protective effect, of which, “Astragalus membranaceus” is a highly valued herb for treatment of cardiovascular and renal diseases in traditional Chinese medicine, In this study, we tested whether calycosin-7-O-β-D-glucoside (Calycosin), a main effective monomer component of “Astragalus membranaceus”, could protect endothelial cells from bacterial endotoxin (LPS)-induced cell injury. Endothelial cell injury was induced by exposing human umbilical vein endothelial cells (HUVECs) to LPS. The effects of calycosin on LPS-induced changes in cell viability, apoptosis rate, cell migration, nitric oxide synthase (NOS), generationof intracellular reactive oxygen species (ROS) and cytoskeleton organization were determined. Microarray assay was employed to screen the possible gene expression change. Based on the results of microarray assay, the expression profile of genes involved in Rho/ROCK pathway and AKT pathway were further evaluated with quantitative real-time RT-PCR or western blot methods. Calycosin improved cell viability, suppressed apoptosis and protected the cells from LPS-induced reduction in cell migration and generation of ROS, protein level of NOS at a comparable magnitude to that of Y27632 and valsartan. Similar to Y27632 and valsartan, Calycosin, also neutralized LPS-induced actomyosin contraction and vinculin protein aggregation. Microarray assay, real-time PCR and western blot results revealed that LPS induced expression of FN, ITG A5, RhoA, PI3K (or PIP2 in western blotting), FAK, VEGF and VEGF R2, and inhibited expression of MLCP. We believed multiple pathways involved in the regulation of calycosin on HUVECs. Calycosin are considered to be able to activate MLCP through promoting the generation of NO, decreasing PMLC, suppressing the cytoskeleton remodeling caused by activation of Rho/ROCK pathway and inhibiting AKT pathway by decreasing VEGF, VEGF R2 and PI3K level. Calycosin protected HUVEC from LPS-induced endothelial injury, possibly through suppression of Rho/ROCK pathway and regulation of AKT pathway.