MicroRNA-24 regulates vascular remodeling via inhibiting PDGF-BB pathway in diabetic rat model
MicroRNA-24 regulates vascular remodeling via inhibiting PDGF-BB pathway in diabetic rat model
复制标题
MicroRNA-24通过抑制PDGF-BB通路调节糖尿病大鼠模型血管重塑
DOI：
10.1016/j.gene.2018.03.056
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发表时间：
2018-06-15
期刊：
影响因子：
3.5
通讯作者：
Chen, Lihua
中科院分区：
文献类型：
--
作者：
Yang, Jian;Zeng, Ping;Chen, Lihua
目的：高血糖是血管成形术所致血管重构的高危因素，而新生内膜增生与颈动脉球囊损伤所致血管重构的发病机制密切相关。研究表明，MicroRNA 24 （miR-24）在心血管疾病中发挥着重要作用，然而，对miR-24的作用的研究还远远不够深入。在本研究中，我们研究了在糖尿病大鼠模型中，通过miR-24重组腺病毒（Ad-miR-24-GFP）上调miR-24是否可以抑制PDGF-BB信号通路并减轻血管重构。方法：雄性sd - dawley大鼠60只，随机分为5组，分别饲喂高糖高脂饲料（Sham组、生理盐水组、Scramble组、Ad-miR-24组）和普通饲料（对照组）。前4组于4周后给予链脲佐菌素（STZ）治疗，密切监测血糖水平。糖尿病大鼠建立成功后，内颈动脉结扎后1.5加压球囊损伤外颈动脉，14d后取血管，检测如下指标：HE染色检测血管内膜厚度水平，免疫组化检测PCNA和P27检测血管平滑肌细胞（VSMCs）增殖程度，qRT-PCR检测miR-24、RAS、PDGF-R水平，western blot检测JNK1/2、p-JNK1/ 2、ERK1/2、p-ERK1/2、RAS、PDGF-R、AP-1、P27和PCNA蛋白水平。血清学检测tnf - α、IL-6、IL-8。结果：将Ad-miR-24递送至球囊损伤部位后，miR-24水平显著升高。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 mRNA表达及蛋白水平。结论：miR-24可通过抑制PDGF-BB信号通路抑制AP-1的表达，从而抑制VSMCs的增殖和血管重构。
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-alpha, 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.