Calreticulin Regulates Neointima Formation and Collagen Deposition following Carotid Artery Ligation.
Calreticulin Regulates Neointima Formation and Collagen Deposition following Carotid Artery Ligation.
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钙网蛋白调节颈动脉连接后的新内膜形成和胶原蛋白沉积。
DOI：
10.1159/000443884
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发表时间：
2015
影响因子：
1.7
通讯作者：
Murphy-Ullrich JE
中科院分区：
文献类型：
--
作者：
Zimmerman KA;Xing D;Pallero MA;Lu A;Ikawa M;Black L;Hoyt KL;Kabarowski JH;Michalak M;Murphy-Ullrich JE
内质网（ER）应激蛋白钙网蛋白（CRT）是TGF-β刺激的成纤维细胞产生细胞外基质（ECM）所必需的。由于TGF-β调节血管纤维增殖反应和胶原沉积，我们研究了CRT敲低对血管平滑肌细胞（VSMCs）纤维增殖反应和胶原沉积的影响。使用血管损伤的颈动脉结扎模型，使用超声将Cre-重组酶-IRES-GFP质粒与微泡（MB）一起递送至CRT floxed小鼠，以特异性地降低颈动脉中的CRT表达。在体外，Cre重组酶介导的CRT敲低在分离的floxed VSMC中减少CRT转录和蛋白，并减弱响应于TGF-β的I型胶原蛋白的诱导。NFAT抑制剂11 R-VIVIT可部分阻断TGF-β对I型胶原的刺激作用。颈动脉结扎后，CRT染色上调，在损伤后14-21天的新生内膜中表达增强。此外，Cre重组酶-IRES-GFP质粒通过靶向超声递送减少CRT floxed小鼠的新生内膜中的CRT表达，并导致新生内膜形成和胶原沉积的显著减少。在局部组织特异性敲除CRT的小鼠中，新生内膜细胞数量也减少。这项工作建立了一个新的作用，CRT介导的VSMC的反应，通过调节胶原沉积和新生内膜的形成。
The endoplasmic reticulum (ER) stress protein, calreticulin (CRT), is required for TGF-β stimulated extracellular matrix (ECM) production by fibroblasts. Since TGF-β regulates vascular fibroproliferative responses and collagen deposition, we investigated the effects of CRT knockdown on vascular smooth muscle cell (VSMCs) fibroproliferative responses and collagen deposition. Using a carotid artery ligation model of vascular injury, Cre-recombinase-IRES-GFP plasmid was delivered with microbubbles (MB) to CRT floxed mice using ultrasound to specifically reduce CRT expression in the carotid artery. In vitro, Cre-recombinase mediated CRT knockdown in isolated floxed VSMCs decreased CRT transcript and protein, and attenuated induction of collagen I protein in response to TGF-β. TGF-β stimulation of collagen I was partly blocked by the NFAT inhibitor 11R-VIVIT. Following carotid artery ligation, CRT staining was upregulated with enhanced expression in the neointima by 14–21 days post-injury. Furthermore, Cre-recombinase-IRES-GFP plasmid delivered by targeted ultrasound reduced CRT expression in the neointima of CRT floxed mice and led to a significant reduction in neointima formation and collagen deposition. Neointimal cell number was also reduced in mice with local, tissue-specific knockdown of CRT. This work establishes a novel role for CRT in mediating VSMC responses to injury through regulation of collagen deposition and neointima formation.