微小 RNA-24 通过抑制糖尿病大鼠模型中的 PDGF-BB 通路调节血管重塑。
MicroRNA-24 regulates vascular remodeling via inhibiting PDGF-BB pathway in diabetic rat model.
机构信息
Department of Cardiology, The First College of Clinical Medical Sciences, China Three Gorges University, Yichang, China; Key Laboratory of Ischemic Cardiovascular and Cerebrovascular Disease Translational Medicine, Three Gorges University, China.
Institute of Cardiovascular Diseases, China Three Gorges University, Yichang, China.
出版信息
Gene. 2018 Jun 15;659:67-76. doi: 10.1016/j.gene.2018.03.056. Epub 2018 Mar 17.
PURPOSE
Hyperglycemia is the high risk factor of vascular remodeling induced by angioplasty, and neointimal hyperplasia is strongly implicated in the pathogenesis of vascular remodeling caused by carotid artery balloon injury. Studies have shown that MicroRNA 24 (miR-24) plays an important role in angiocardiopathy, However, the role of miR-24 is far from thorough research. In this study, we investigate whether up-regulation of miR-24 by using miR-24 recombinant adenovirus (Ad-miR-24-GFP) can inhibit PDGF-BB signaling pathway and attenuate vascular remodeling in the diabetic rat model.
METHODS
Male Sprague-Dawley rats (n = 60) were randomly divided into 5 groups and fed with high sugar and high fat diet (Sham, Saline, Scramble, Ad-miR-24 groups), or ordinary diet (Control group). The front four groups were treated with streptozotocin (STZ) four weeks later and the blood glucose level was closely monitored. After the successful establishment of diabetic rats, the external carotid artery was injured by pressuring balloon 1.5 after internal carotid artery ligation, then the blood vessels were harvested 14 days later and indexes were detected including the following: HE staining for the level of vascular intima thickness, immunohistochemical detection for PCNA and P27 to test the proliferative degree of vascular smooth muscle cells (VSMCs), qRT-PCR for the level of miR-24, RAS,PDGF-R, western blot for the protein levels of JNK1/2, p- JNK1/2, ERK1/2, p-ERK1/2, RAS, PDGF-R, AP-1,P27 and PCNA. Serological detection was conducted for TNF-α, IL-6, IL-8.
RESULTS
The delivery of Ad-miR-24 into balloon injury site has significantly increased the level of miR-24. Up-regulation of miR-24 could regulate vascular remodeling effectively, lower the level of inflammatory factors, inhibit the expression of mRNA and protein levels of JNK1/2, ERK1/2, RAS, PDGF-R, AP-1, P27, PCNA.
CONCLUSION
miR-24 can inhibit the expression of AP-1 via the inhibition of PDGF-BB signaling pathway, thus inhibit VSMCs proliferation and vascular remodeling.
目的
高血糖是血管成形术后血管重构的高危因素，而血管平滑肌细胞（VSMC）的增殖在血管重构中起关键作用。球囊损伤后的颈动脉内膜增生强烈提示血管重构与血管损伤后细胞外基质的降解和合成失衡有关。研究表明 MicroRNA 24（miR-24）在心血管疾病中起重要作用，然而，miR-24 的作用还远未被彻底研究。本研究通过 miR-24 重组腺病毒（Ad-miR-24-GFP）上调 miR-24，观察其对糖尿病大鼠模型 PDGF-BB 信号通路的抑制作用及对血管重构的影响。
方法
雄性 Sprague-Dawley 大鼠（n=60）随机分为 5 组，分别给予高糖高脂饮食（假手术组、盐水组、 scramble 组、Ad-miR-24 组）或普通饮食（对照组）。前 4 组在 4 周后给予链脲佐菌素（STZ）处理，并密切监测血糖水平。成功建立糖尿病大鼠模型后，结扎颈内动脉的同时对内颈动脉进行球囊压迫损伤，1.5 周后取血管标本，检测指标包括：HE 染色检测血管内膜厚度，免疫组化检测增殖细胞核抗原（PCNA）和 P27 以检测 VSMC 的增殖程度，qRT-PCR 检测 miR-24 水平，RAS、PDGF-R，Western blot 检测 JNK1/2、p-JNK1/2、ERK1/2、p-ERK1/2、RAS、PDGF-R、AP-1、P27 和 PCNA 的蛋白水平。检测血清 TNF-α、IL-6、IL-8。
结果
Ad-miR-24 递送至球囊损伤部位可显著增加 miR-24 水平。上调 miR-24 可有效调节血管重构，降低炎症因子水平，抑制 JNK1/2、ERK1/2、RAS、PDGF-R、AP-1、P27、PCNA 的 mRNA 和蛋白水平表达。
结论
miR-24 通过抑制 PDGF-BB 信号通路抑制 AP-1 的表达，从而抑制 VSMC 的增殖和血管重构。
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