MicroRNA-133a impairs perfusion recovery after hindlimb ischemia in diabetic mice
MicroRNA-133a impairs perfusion recovery after hindlimb ischemia in diabetic mice
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MicroRNA-133a 损害糖尿病小鼠后肢缺血后的灌注恢复
DOI：
10.1042/bsr20180346
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发表时间：
2018-08-31
影响因子：
4
通讯作者：
Lu, Wenju
中科院分区：
文献类型：
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作者：
Chen, Lingdan;Liu, Chunli;Lu, Wenju
目的：糖尿病周围动脉病变(PAD)患者缺血后新生血管受损，预后较差。在糖尿病条件下，microRNA(MiR)-133a在血管内皮细胞中过度表达。在这里，我们测试糖尿病诱导的miR-133a上调是否与实验性PAD模型的新生血管能力受损有关。方法和结果：用定量RT-PCR方法检测MIR-133a在糖尿病小鼠缺血肌肉中的表达水平，结果表明糖尿病小鼠缺血肌肉中MIR-133a的表达水平高于非糖尿病小鼠。使用antagomir基因敲除miR-133a可改善糖尿病合并HLI后第21天实验性PAD模型的血流灌注恢复和血管生成。另一方面，miR-133a的过度表达损害了血流灌注的恢复。实验PAD后第7天取材进行生化检测，miR-133a拮抗剂可降低丙二醛含量，升高GTP环水解酶1(GCH1)和环鸟嘌呤单磷酸(CGMP)水平。在培养的内皮细胞中，miR-133a拮抗可导致内皮细胞内氧自由基水平降低，并增加血管形成、一氧化氮(NO)和cGMP水平。此外，GCH1抑制剂可阻断miR-133a拮抗剂诱导的血管生成。相反，在非糖尿病模型中，miR-133a过表达会损害血管生成，并降低GCH1、NO和cGMP水平。结论：糖尿病诱导的miR-133a上调通过减少血管内皮细胞中NO的合成而损害PAD的血管生成。MIR-133a拮抗剂可促进缺血后血管生成。
Objective: Peripheral arterial disease (PAD) patients with diabetes mellitus suffer from impaired neovascularization after ischemia which results in poorer outcomes. MicroRNA (miR)-133a is excessively expressed in endothelial cells under diabetic conditions. Here, we test whether diabetes-induced miR-133a up-regulation is involved in the impaired capability of neovascularization in experimental PAD models. Methods and results: MiR-133a level was measured by quantitative RT-PCR and showed a higher expression level in the ischemic muscle from diabetic mice when compared with nondiabetic mice. Knockdown of miR-133a using antagomir improved perfusion recovery and angiogenesis in experimental PAD model with diabetes day 21 after HLI. On the other hand, overexpression of miR-133a impaired perfusion recovery. Ischemic muscle was harvested day 7 after experimental PAD for biochemical test, miR-133a antagonism resulted in reduced malondialdehyde, and it increased GTP cyclohydrolase 1 (GCH1), and cyclic guanine monophosphate (cGMP) levels. In cultured endothelial cells, miR-133a antagonism resulted in reduced reactive oxygen species level, and it increased tube formation, nitric oxide (NO), and cGMP level. Moreover, miR-133a antagonism-induced angiogenesis was abolished by GCH1 inhibitor. In contrary, miR-133a overexpression impairs angiogenesis and it reduces GCH1, NO, and cGMP levels in nondiabetic models. Conclusion: Diabetes mellitus-induced miR-133a up-regulation impairs angiogenesis in PAD by reducing NO synthesis in endothelial cells. MiR-133a antagonism improves postischemic angiogenesis.