The deregulation of STIM1 and store operative calcium entry impaired aortic smooth muscle cells contractility in aortic medial degeneration
The deregulation of STIM1 and store operative calcium entry impaired aortic smooth muscle cells contractility in aortic medial degeneration
复制标题
STIM1 的失调和存储操作性钙进入损害了主动脉内侧变性中的主动脉平滑肌细胞收缩力
DOI：
10.1042/bsr20181504
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发表时间：
2019-01-31
影响因子：
4
通讯作者：
Wang, Zhiwei
中科院分区：
文献类型：
--
作者：
Hong, Junmou;Hu, Zhipeng;Wang, Zhiwei
背景资料：临床主动脉样本的微阵列分析表明，基质相互作用分子1（STIM 1）在调节主动脉中层变性（AMD）中的潜在作用，尽管STIM 1在正常主动脉平滑肌细胞（ASMC）中的不确定性。本研究旨在探讨STIM 1在AMD中的表达变化及其可能的机制。方法：采用ApoE−/−小鼠自体注射血管紧张素II（Ang II）建立AMD模型。我们评估了STIM 1抑制剂SKF 96365对AMD模型和体外培养ASMCs的影响。弹性货车Gieson（EVG）染色用于可视化弹性纤维损伤。透射电镜观察线粒体变化。通过在流式细胞仪中测量fluo-4染色来定量细胞质钙。机械拉伸装置用于模拟ASMCs在体内经历的拉伸。Annexin V/PI染色检测细胞凋亡;采用免疫印迹、免疫组化和免疫荧光法检测STIM 1、收缩相关蛋白（α-平滑肌肌动蛋白（α-SMA）、肌球蛋白轻链（MLC））、内质网（ER）应激相关蛋白（CHOP、转录激活因子6（ATF-6））和smad 2/3的表达。结果：SKF 96365可加重AMD模型主动脉损伤。SKF 96365可降低ASMCs胞浆钙浓度，引起线粒体肿胀，并增加ATF-6和CHOP的表达。SKF 96365可降低平滑肌细胞MLC和α-SMA的表达，使平滑肌细胞对机械牵张敏感。SKF 96365可抑制转化生长因子β1（TGFβ1）处理后smad 2/3的活化。结论：STIM 1在ASMCs中是不可缺少的。干预STIM 1可通过调节收缩蛋白的表达，诱导ASMCs的ER应激，从而加重AMD的进程。
Background: Microarray analysis of clinical aortic samples suggested a potential role for stromal interaction molecule 1 (STIM1) in the modulation of aortic medial degeneration (AMD), despite the uncertainty about STIM1 in normal aortic smooth muscle cells (ASMCs). Here, we aimed to explore changes in STIM1 expression in AMD, and the possible mechanisms. Methods: An AMD model was established using auto-delivery of angiotensin II (Ang II) into ApoE−/− mice. We assessed the effects of SKF96365, a STIM1 inhibitor, in AMD model and in vitro cultured ASMCs. Elastic van Gieson (EVG) staining was used to visualize elastic fiber injury. Mitochondria changes were viewed by TEM. Cytoplasmic calcium was quantified by measuring fluo-4 staining in a flow cytometer. Mechanical stretching device was used to mimic stretching that ASMCs experience in vivo. Cell apoptosis was determined by using Annexin V/propidium iodide (PI) staining. The expression of STIM1, contractile related proteins (α-smooth muscle actin (α-SMA), myosin light chain (MLC)), endoplasmic reticulum (ER) stress-related proteins (CHOP, activating transcription factor 6 (ATF-6)) and smad2/3 were assessed by Western blotting, immunohistochemistry (IHC), and immunofluorescence (IF). Results: SKF96365 exacerbated aortic injury in the AMD model. SKF96365 reduced cytoplasmic calcium concentration in ASMCs, caused mitochondrial swelling, and elevated the expression of ATF-6 and CHOP. SKF96365 decreased the expression of MLC and α-SMA in ASMCs, causing them to be vulnerable to mechanical stretch. SKF96365 suppressed smad2/3 activation after treatment with transforming growth factor (TGF) β1 (TGFβ1). Conclusions: STIM1 is indispensable in ASMCs. Interfering with STIM1 exaggerated the AMD process by modulating the expression of contractile proteins, inducing ER stress in ASMCs.