To investigate the protective effect of inhibiting miR-181a on diabetic corneal nerve in mice, we chose male C57BL/6 mice with streptozotocin (STZ) -induced diabetes as animal models. The expression of miR-181a in trigeminal ganglion tissue (TG) of diabetic mice was detected by real-time PCR. In vitro, we cultured mouse trigeminal ganglion neurons and measured the neuronal axon growth when treated under miR-181a antagomir and negative conditions (NTC). Immunofluorescence showed a significant increase in neuronal axon length in trigeminal ganglion cells treated with miR-181a antagomir. In animal models, we performed epithelial scraping and subconjunctival injection of the miR-181a antagomir and miRNA antagomir NTC to observe the corneal nerve repair by corneal nerve staining. miR-181a antagomir subconjunctival injection significantly increased the corneal epithelium healing of diabetic mice compared with that of the NTC group. Meanwhile, corneal nerve staining showed that the repair of corneal nerve endings was significantly promoted. As the targets of the 181a, ATG5 and BCL-2 were previously identified. The results of Western blot showed that the expression of autophagy associated protein ATG5 and LC3B-II and the expression of anti-apoptotic protein Bcl-2 were decreased in the high-glucose cell culture environment and the diabetic TG tissue. The expression of ATG5, LC3B-II and Bcl-2 were significantly increased after miR-181a antagomir treatment compared with negative control group. This study showed that inhibition of miR-181a expression in diabetic mice could increase ATG5-mediated autophagic activation, BCL-2-mediated inhibition of apoptosis, and promote the growth of trigeminal sensory neurons and the regeneration of corneal nerve fibers. It has a protective effect on diabetic corneal neuropathy.

中文翻译：

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在小鼠模型中，保护作用抑制了miR-181a在糖尿病角膜神经上的表达。

为了研究抑制miR-181a对小鼠糖尿病性角膜神经的保护作用，我们选择了链脲佐菌素（STZ）诱导的糖尿病的雄性C57BL / 6小鼠作为动物模型。通过实时PCR检测miR-181a在糖尿病小鼠三叉神经节组织（TG）中的表达。在体外，我们培养了小鼠三叉神经节神经元，并在miR-181a antagomir和阴性条件（NTC）处理下测量了神经元轴突的生长。免疫荧光显示，用miR-181a antagomir处理的三叉神经节细胞中神经元轴突长度显着增加。在动物模型中，我们进行了miR-181a antagomir和miRNA antagomir NTC的上皮刮擦和结膜下注射，以通过角膜神经染色观察角膜神经修复。与NTC组相比，miR-181a antagomir结膜下注射显着增加了糖尿病小鼠的角膜上皮愈合。同时，角膜神经染色显示角膜神经末梢的修复得到显着促进。作为181a的靶标，以前已经确定了ATG5和BCL-2。Western blot结果显示，在高糖细胞培养环境和糖尿病TG组织中，自噬相关蛋白ATG5和LC3B-II的表达和抗凋亡蛋白Bcl-2的表达均降低。与阴性对照组相比，miR-181a antagomir处理后ATG5，LC3B-II和Bcl-2的表达明显增加。这项研究表明，抑制糖尿病小鼠中miR-181a的表达可以增加ATG5介导的自噬激活，BCL-2介导的凋亡抑制，并促进三叉神经感觉神经元的生长和角膜神经纤维的再生。它对糖尿病性角膜神经病具有保护作用。