目的:本研究旨在探讨microRNA(miR)-193a促进早期糖尿病(DM)大鼠视网膜神经细胞凋亡的作用及其机制。
方法:选择72只雄性SD级大鼠腹腔注射链脲佐菌素(STZ)建立DM模型,并随机分为对照组(空白对照组),糖尿病组(糖尿病模型组),DM+miR-NC抑制剂组(miR-193a抑制阴性对照组),DM+miR-193a抑制剂组(miR-193a抑制剂组),DM+miR-NC模拟物组(miR-193a过表达阴性对照组),DM+miR-193a模拟组(miR-193a过表达组),每组12只大鼠。
结果:miR-193a表达,凋亡率,还有Bax,Caspase3和Caspase9蛋白表达水平升高,Bcl-2蛋白在DM大鼠视网膜组织和高糖诱导的大鼠视网膜神经元细胞中的表达降低,而miR-193a抑制剂逆转了这些过程。这些双荧光素酶报告分析表明WT1CDS,和WT1Mut在miR-193a组低于miR-NC组(P<0.05);WT1蛋白在DM大鼠视网膜组织和高糖诱导的大鼠视网膜神经元细胞中表达降低,和miR-193a抑制剂增加WT1蛋白表达。与miR-193a和WT1载体共转染的细胞相比,miR-193a和WT1共转染抑制高糖诱导的视网膜神经元细胞凋亡并调节凋亡蛋白表达。miR-193a在DM大鼠视网膜组织和高糖诱导的大鼠视网膜神经元细胞中高表达,WT1低表达。
结论:miR-193a可能通过靶向和负调控WT1表达来抑制DM大鼠早期视网膜神经细胞凋亡。
OBJECTIVE: This study aimed to investigate the role and mechanism of microRNA (miR)-193a in promoting apoptosis of retinal neuronal cells in early diabetic (DM) rats.
METHODS: Seventy-two male SD-grade rats were selected to establish a DM model by intraperitoneal injection of streptozotocin (STZ), and randomly divided into a control group (blank control group), a DM group (diabetic model group), a DM+miR-NC inhibitor group (miR-193a inhibition negative control group), a DM+miR-193a inhibitor group (miR-193a inhibitor group), DM+miR-NC mimic group (miR-193a overexpression negative control group), DM+miR-193a mimic group (miR-193a overexpression group), with12 rats in each group.
RESULTS: The miR-193a expression, apoptosis rate, and Bax, Caspase3, and Caspase9 protein expression levels were elevated, and Bcl-2 protein expression was decreased in the retinal tissues of DM rats and high glucose-induced rat retinal neuronal cells, while miR-193a inhibitors reversed these processes. These dual luciferase reporter assay showed that WT1CDS, and WT1Mut were lower in the miR-193a group than in the miR-NC group (P<0.05); WT1 protein expression was reduced in the retinal tissues of DM rat and high glucose-induced rat retinal neuronal cells, and miR-193a inhibitors increased WT1 protein expression. Compared with cells co-transfected with miR-193a and WT1vector, miR-193a and WT1 cotransfection inhibited high glucose-induced apoptosis in retinal neuronal cells and regulated apoptotic protein expression. miR-193a was highly expressed and WT1 was lowly expressed in retinal tissues of DM rats and high glucose-induced rat retinal neuronal cells.
CONCLUSIONS: miR-193a could inhibit early retinal neuronal cell apoptosis in DM rats by targeting and negatively regulating WT1 expression.