摘要
目的探究miR-199-3p在糖尿病视网膜病变(DR)中的作用及其机制。方法单次注射链脲佐菌素建立大鼠糖尿病模型,RT-qPCR检测大鼠视网膜组织miR-199-3p、血管内皮生长因子A(VEGFA)表达;双荧光素实验验证miR-199-3p和VegfA的靶向关系;TUNEL染色检测视网膜细胞凋亡;Western blot检测VEGFA、转化生长因子-β(TGF-β)、肝细胞生长因子(HGF)、色素上皮衍生因子(PEDF)蛋白。结果成功建立糖尿病模型大鼠,且模型大鼠视网膜组织miR-199-3p显著下降,VEGFA显著升高(P<0.001);miR-199-3p靶向VegfA。miR-199-3p抑制显著降低视网膜神经节细胞数目,增强视网膜细胞凋亡,而抑制VegfA可显著减轻miR-199-3p抑制对视网膜细胞的损伤(P<0.01)。miR-199-3p过表达后TGF-β1、HGF显著降低,PEDF升高(P<0.001),而过表达miR-199-3p联合VEGFA可显著改变这种作用。结论miR-199-3p通过靶向VegfA抑制糖尿病大鼠视网膜病变。
Objective To study the role and mechanism of mir-199-3p in the pathogenesis of diabetic retinopathy(DR).Methods The diabetic model of rats was established by injection of streptozotocin.The expression of miR-199-3p and vascular endothelial growth factor A(VEGFA)in the retina of rats was detected by RT-qPCR.The targeting relationship between miR-199-3p and VegfA in retinal cells was studied by dual luciferase assays.Apoptosis of retinal cells was evaluated by TUNEL staining.Protein of VEGFA and transforming growth factor-β(TGF-β),Hepatocyte growth factor(HGF),pigment epithelium derived factor(PEDF)were detected by Western blot.Results The retinal staining showed that the pathological changes such as retinal capillary dilation and vascular curvature in diabetic rats.The results of reverse transcription showed that the expression of miR-199-3p decreased and VEGFA increased in the retina of diabetic rats significantly.The targeting relationship between miR-199-3p and VegfA was confirmed by dual luciferase assays.The low-expression of miR-199-3p significantly reduced the number of retinal ganglion cells and enhance the apoptosis of retinal cells,while the low expression-miR-199-3p and VEGFA significantly reversed the effects of miR-199-3p on retinal cells(P<0.01).Western blot results showed that the expression of TGF-βand HGF decreased significantly after over-expression of miR-199-3p with the increased expression of PEDF(P<0.001).However,over-expression of miR-199-3p combined with VEGFA expression significantly reversed the expression of these proteins.Conclusions miR-199-3p inhibits diabetic retinopathy by targeting at VegfA.
作者
周悦
林静娜
ZHOU Yue;LIN Jingna(Hospital,the Joint Logistic Support Force,Zhangzhou 363000,China)
出处
《基础医学与临床》
2023年第7期1084-1089,共6页
Basic and Clinical Medicine
基金
福建医学科技青年培育项目(18FBQN2016017)。
