Neurosurgery, The Affiliated Hospital of Qingdao University, 16 Jiangsu Road, Qingdao, China.
Institute of Neuroregeneration & Neurorehabilitation, Department of Pathophysiology, Qingdao University, 308 Ningxia Road, Qingdao, China.
J Stroke Cerebrovasc Dis. 2022 Oct;31(10):106670. doi: 10.1016/j.jstrokecerebrovasdis.2022.106670. Epub 2022 Aug 13.
To examine the role of CCL14 in the neovascularization process and vulnerability progression within carotid plaques by investigating the mechanism of CCL14 regulation of VEGF-A.
We first performed histological analysis and immunohistochemical staining of human carotid plaque tissue to detect the expression of CCL14, JAK2, STAT3 and VEGF-A. We next examined the protein expression of CCL14, VEGF-A, JAK2, STAT3, and phosphorylation of JAK2 and STAT3 in human carotid atherosclerotic plaques by Western blotting. Finally, we performed in vitro culture of human umbilical vein endothelial cells (HUVEC). In the tube formation assay of HUVEC, we added CCL14 siRNA or VEGF-A siRNA to the culture medium using lentiviral transfection to knock down CCL14 or VEGF-A and grouped them for control assays, and detected the changes in the expression of the above proteins using Western blotting.
Histological and Western blotting analysis of human carotid plaque samples showed that the expression of CCL14 and VEGF-A was higher in the vulnerable plaques than in stable plaques. In the in vitro cultures of HUVEC, CCL14 was found to increase the number and length of intercellularly generated tubular structures. CCL14 increases VEGF-A expression via activating JAK2/STAT3 signaling.
In the human carotid plaques, CCL14 promotes angiogenesis by upregulation of VEGF-A via JAK2/STAT3 pathway and thus drives the progression of carotid plaques vulnerability.
通过研究 CCL14 对 VEGF-A 的调节机制，探讨 CCL14 在颈动脉斑块内新生血管形成过程和易损性进展中的作用。
我们首先对人颈动脉斑块组织进行组织学分析和免疫组织化学染色，以检测 CCL14、JAK2、STAT3 和 VEGF-A 的表达。然后，我们通过 Western blot 检测人颈动脉粥样硬化斑块中 CCL14、VEGF-A、JAK2、STAT3 的蛋白表达以及 JAK2 和 STAT3 的磷酸化。最后，我们进行了人脐静脉内皮细胞（HUVEC）的体外培养。在 HUVEC 的管形成试验中，我们通过慢病毒转染将 CCL14 siRNA 或 VEGF-A siRNA 添加到培养基中，以敲低 CCL14 或 VEGF-A，并将其分组进行对照试验，然后使用 Western blot 检测上述蛋白表达的变化。
人颈动脉斑块样本的组织学和 Western blot 分析显示，易损斑块中 CCL14 和 VEGF-A 的表达高于稳定斑块。在 HUVEC 的体外培养中，发现 CCL14 增加了细胞间生成的管状结构的数量和长度。CCL14 通过激活 JAK2/STAT3 信号通路增加 VEGF-A 的表达。
在人颈动脉斑块中，CCL14 通过 JAK2/STAT3 通路上调 VEGF-A 促进血管生成，从而推动颈动脉斑块易损性的进展。
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