MicroRNA-320 EXPRESSION IN MYOCARDIAL MICROVASCULAR ENDOTHELIAL CELLS AND ITS RELATIONSHIP WITH INSULIN-LIKE GROWTH FACTOR-1 IN TYPE 2 DIABETIC RATS
MicroRNA-320 EXPRESSION IN MYOCARDIAL MICROVASCULAR ENDOTHELIAL CELLS AND ITS RELATIONSHIP WITH INSULIN-LIKE GROWTH FACTOR-1 IN TYPE 2 DIABETIC RATS
复制标题
2型糖尿病大鼠心肌微血管内皮细胞中MicroRNA-320的表达及其与胰岛素样生长因子1的关系
DOI：
10.1111/j.1440-1681.2008.05057.x
复制
发表时间：
2009-02-01
影响因子：
2.9
通讯作者：
Hu, R. M.
中科院分区：
文献类型：
--
作者：
Wang, X. H.;Qian, R. Z.;Hu, R. M.
本研究的目的是确定心肌微血管内皮细胞 (MMVEC) 在 2 型糖尿病 Goto-Kakizaki (GK) 大鼠血管生成受损中的作用。使用 microRNA (miRNA) 微阵列评估 GK 和 Wistar 大鼠 MMVEC 中的 miRNA 表达。使用实时逆转录聚合酶链反应 (RT-PCR) 在 GK 大鼠的 MMVEC 中观察到 miRNA-320 的上调。到目前为止，已报道 9 种 miRNA 靶向血管生成因子和/或受体，包括含有激酶插入结构域的受体 (Flk-1)、胰岛素样生长因子 1 (IGF-1) 和胰岛素样生长因子 1 受体 (IGF-1R)。预测miR-320靶向的基因包括Flk-1、IGF-1和IGF-1R。 Western blot分析和RT-PCR分别用于分析推定基因IGF-1和IGF-1R的蛋白质和mRNA表达。糖尿病MMVEC中IGF-1和IGF-1R蛋白的表达显着降低。然而，IGF-1 mRNA 的表达增加而不是减少。糖尿病和对照MMVEC之间IGF-1R的mRNA表达没有显着差异。将miR-320抑制剂转染到GK大鼠的MMVEC中证实miR-320损害血管生成。转染 miR-320 抑制剂后，糖尿病 MMVEC 的增殖和迁移得到改善。此外，miR-320抑制剂显着增加了IGF-1蛋白的表达，但对IGF-1R的表达没有影响。与Wistar大鼠相比，GK大鼠的MMVEC中有11个miRNA上调：let-7e、miR-129、miR-291-5p、miR-320、miR-327、mir-333、miR-363-5p、 miR-370、miR-494、miR-503 和 miR-664。结果表明，GK 大鼠 MMVEC 中 miR-320 的上调可能是 IGF-1 蛋白和 mRNA 表达不一致的原因，因此与糖尿病血管生成受损有关。 miR-320抑制剂的转染可能是治疗糖尿病血管生成受损的一种治疗方法。
The aim of the present study was to determine the role of myocardial microvascular endothelial cells (MMVEC) in impaired angiogenesis of type 2 diabetic Goto-Kakizaki (GK) rats.A microRNA (miRNA) microarray was used to assess miRNA expression in MMVEC from GK and Wistar rats. Upregulation of miRNA-320 was observed in MMVEC from GK rats using real-time reverse transcription-polymerase chain reaction (RT-PCR).So far, nine miRNAs have been reported to target angiogenic factors and/or receptors, including kinase insert domain containing receptor (Flk-1), insulin-like growth factor 1 (IGF-1) and insulin-like growth factor 1 receptor (IGF-1R). The predicted genes targeted by miR-320 include Flk-1, IGF-1 and IGF-1R. Western blot analysis and RT-PCR were used to analyse the protein and mRNA expression, respectively, of the putative genes IGF-1 and IGF-1R. The expression of IGF-1 and IGF-1R proteins decreased significantly in diabetic MMVEC. However, the expression of IGF-1 mRNA increased rather than decreased. The mRNA expression of IGF-1R did not differ significantly between diabetic and control MMVEC.Transfection of an miR-320 inhibitor into MMVEC from GK rats confirmed that miR-320 impaired angiogenesis. The proliferation and migration of diabetic MMVEC improved after transfection of the miR-320 inhibitor. In addition, the miR-320 inhibitor significantly increased the expression of IGF-1 protein, but had no effect on the expression of IGF-1R.Eleven miRNAs were upregulated in MMVEC from GK rats compared with those in Wistar rats: let-7e, miR-129, miR-291-5p, miR-320, miR-327, mir-333, miR-363-5p, miR-370, miR-494, miR-503 and miR-664.The results indicate that upregulation of miR-320 in MMVEC from GK rats may be responsible for the inconsistency between the expression of IGF-1 protein and mRNA and therefore related to impaired angiogenesis in diabetes. Transfection of an miR-320 inhibitor may be a therapeutic approach for the treatment of impaired angiogenesis in diabetes.