Narayanan Sampath, Röhl Samuel, Lengquist Mariette, Kronqvist Malin, Matic Ljubica, Razuvaev Anton
Vascular Surgery, Department of Molecular Medicine and Surgery, Karolinska University Hospital and Karolinska Institutet, Stockholm, Sweden.
JVS Vasc Sci. 2023 May 19;4:100111. doi: 10.1016/j.jvssci.2023.100111. eCollection 2023.
Atherosclerosis is a leading cause of mortality in the rapidly growing population with diabetes mellitus. Vascular interventions in patients with diabetes can lead to complications attributed to defective vascular remodeling and impaired healing response in the vessel wall. In this study, we aim to elucidate the molecular differences in the vascular healing response over time using a rat model of arterial injury applied to healthy and diabetic conditions.
Wistar (healthy) and Goto-Kakizaki (GK, diabetic) rats (n = 40 per strain) were subjected to left common carotid artery (CCA) balloon injury and euthanized at different timepoints: 0 and 20 hours, 5 days, and 2, 4, and 6 weeks. Noninvasive morphological and physiological assessment of the CCA was performed with ultrasound biomicroscopy (Vevo 2100) and corroborated with histology. Total RNA was isolated from the injured CCA at each timepoint, and microarray profiling was performed (n = 3 rats per timepoint; RaGene-1_0-st-v1 platform). Bioinformatic analyses were conducted using R software, DAVID bioinformatic tool, online STRING database, and Cytoscape software.
Significant increase in the neointimal thickness ( < .01; two-way analysis of variance) as well as exaggerated negative remodeling was observed after 2 weeks of injury in GK rats compared with heathy rats, which was confirmed by histological analyses. Bioinformatic analyses showed defective expression patterns for smooth muscle cells and immune cell markers, along with reduced expression of key extracellular matrix-related genes and increased expression of pro-thrombotic genes, indicating potential faults on cell regulation level. Transcription factor-protein-protein interaction analysis provided mechanistic evidence with an array of transcription factors dysregulated in diabetic rats.
In this study, we have demonstrated that diabetic rats exhibit impaired arterial remodeling characterized by a delayed healing response. We show that increased contractile smooth muscle cell marker expression coincided with decreased matrix metalloproteinase expression, indicating a potential mechanism for a lack of extracellular matrix reorganization in the impaired vascular healing in GK rats. These results further corroborate the higher prevalence of restenosis in patients with diabetes and provide vital molecular insights into the mechanisms contributing to the impaired arterial healing response in diabetes. Moreover, the presented study provides the research community with the valuable longitudinal gene expression data bank for further exploration of diabetic vasculopathy.
在快速增长的糖尿病患者群体中，动脉粥样硬化是主要的死亡原因。对糖尿病患者进行血管干预可能会导致并发症，这归因于血管重塑缺陷和血管壁愈合反应受损。在本研究中，我们旨在使用应用于健康和糖尿病状态的动脉损伤大鼠模型，阐明血管愈合反应随时间的分子差异。
将Wistar（健康）大鼠和Goto-Kakizaki（GK，糖尿病）大鼠（每个品系n = 40）进行左颈总动脉（CCA）球囊损伤，并在不同时间点实施安乐死：0和20小时、5天、2、4和6周。使用超声生物显微镜（Vevo 2100）对CCA进行非侵入性形态学和生理学评估，并通过组织学进行验证。在每个时间点从受伤的CCA中分离总RNA，并进行微阵列分析（每个时间点n = 3只大鼠；RaGene-1_0-st-v1平台）。使用R软件、DAVID生物信息工具、在线STRING数据库和Cytoscape软件进行生物信息学分析。
与健康大鼠相比，在损伤2周后，GK大鼠的新生内膜厚度显著增加（P <.01；双向方差分析），并且观察到过度的负向重塑，这通过组织学分析得到证实。生物信息学分析显示平滑肌细胞和免疫细胞标志物的表达模式存在缺陷，同时关键细胞外基质相关基因的表达降低，促血栓形成基因的表达增加，表明在细胞调节水平上存在潜在缺陷。转录因子-蛋白质-蛋白质相互作用分析提供了机制证据，表明一系列转录因子在糖尿病大鼠中失调。
在本研究中，我们证明糖尿病大鼠表现出动脉重塑受损，其特征为愈合反应延迟。我们表明收缩性平滑肌细胞标志物表达增加与基质金属蛋白酶表达降低同时出现，这表明GK大鼠血管愈合受损时细胞外基质重组缺乏的潜在机制。这些结果进一步证实了糖尿病患者再狭窄的较高患病率，并为糖尿病患者动脉愈合反应受损的机制提供了重要的分子见解。此外，本研究为研究界提供了宝贵的纵向基因表达数据库，以供进一步探索糖尿病血管病变。
