Department of Biochemistry and Molecular Biology, Binzhou Medical University, YanTai, ShanDong, People's Republic of China.
Mol Cell Biochem. 2012 Feb;361(1-2):305-14. doi: 10.1007/s11010-011-1116-7. Epub 2011 Oct 20.
To characterize the roles of tribble 2 (TRB2) and its targeted microRNAs (miRNAs) in the pathogenesis of the early vascular injury involved in diabetic-2 rat. Goto-Kakizaki (GK) rat and Wistar rat were used as the animal models. Each eligible rat was killed and the rat aorta tissues were analyzed by immunohistochemistry, ELISA, reverse transcription-polymerase chain reaction (RT-PCR), and real-time PCR detection. GFP expression in RAOEC cells (rat vascular aortic endothelial cell)were detected by flow cytometry and fluorescent microscope. TRB2 gene expression was increased in endothelia cell and the adventitia of Goto-Kakizaki (GK) rat compared with Wistar rat. Next, studies using RAOEC cells showed that the TRB2 expression was inhibited by the treatment of miR-98. We further showed that the expression of miR-98 was significantly decreased in the adventitia and endomembrane at different degrees in GK rats compared with control. Finally, we validated the changes in TRB2 by studying one of the TRB2's substrates, Akt, in animal models. We expected a corresponding change in the levels of phosphorylated Akt. Indeed, our results showed that the phosphorylation of Akt at Thr 308 in the endothelial cells and phosphorylation of Akt at Ser 473 in adventitia was decreased in GK rats, compared with Wistar control. TRB2 plays important roles in the pathogenesis of diabetic-2 large artery complications at early stage, and these effects may be modulated by miR-98. Thus, targeting TRB2 and miR-98 could be considered as novel therapeutic strategies for the early large artery deficits in diabetic-2.
为了研究 tribble 2（TRB2）及其靶向 microRNAs（miRNAs）在涉及糖尿病 2 型大鼠早期血管损伤发病机制中的作用，使用 Goto-Kakizaki（GK）大鼠和 Wistar 大鼠作为动物模型。处死每个合格的大鼠，通过免疫组织化学、ELISA、逆转录-聚合酶链反应（RT-PCR）和实时 PCR 检测分析大鼠主动脉组织。通过流式细胞术和荧光显微镜检测 RAOEC 细胞（大鼠血管主动脉内皮细胞）中的 GFP 表达。与 Wistar 大鼠相比，在 GK 大鼠的内皮细胞和外膜中 TRB2 基因表达增加。接下来，使用 RAOEC 细胞的研究表明，miR-98 的处理抑制了 TRB2 的表达。我们进一步表明，与对照组相比，GK 大鼠的外膜和内膜中 miR-98 的表达程度不同，均显著降低。最后，我们通过研究动物模型中 TRB2 的一种底物 Akt 来验证 TRB2 的变化。我们期望 Akt 的磷酸化水平会相应改变。事实上，我们的结果表明，与 Wistar 对照组相比，内皮细胞中 Akt 的 Thr 308 磷酸化和外膜中 Akt 的 Ser 473 磷酸化在 GK 大鼠中减少。TRB2 在糖尿病 2 型大血管并发症的早期发病机制中发挥重要作用，这些作用可能由 miR-98 调节。因此，靶向 TRB2 和 miR-98 可以被认为是糖尿病 2 型早期大血管缺陷的新治疗策略。
