miR-28-3p通过PI3K/AKT通路对大鼠糖尿病性视网膜病变及视网膜血管生成的影响  

Effect of miR-28-3p via PI3K/AKT pathway on diabetic retinopathy and retinal angiogenesis in rats

作　　者：闫欢欢 李春花[1] 曾戎[1] 陈红 苏伟[2] Huanhuan Yan;Chunhua Li;Rong Zeng;Hong Chen;WeiSu(Department of Ophthalmology,Xi’an Fourth Hospital,Xi’an 710004,China;School of Basic Medicine,Xi’an Jiaotong University,Xi’an 710049,China)

机构地区：[1]陕西省西安市第四医院眼科,西安710004 [2]西安交通大学基础医学院,西安710049

出　　处：《广西医科大学学报》2020年第8期1389-1397,共9页Journal of Guangxi Medical University

基　　金：supported by Key Research & Development Projects in Shanxi Province (N0.2019SF-126).

摘　　要：目的:探讨miR-28-3p通过PI3K/AKT通路对糖尿病性视网膜病变(DR)大鼠及视网膜血管生成的影响。方法:SD大鼠分为假手术组(sham组)、糖尿病模型组(model组)、糖尿病+阴性对照组(miR-NC组)和糖尿病+miR-28-3p inhibitor组(miR-28-3p inhibitor组),采用腹腔注射链脲佐菌素溶液建立糖尿病大鼠模型并给与腺相关病毒(AAV-U6-inhibitor-rno-miR-28-3p-CAG-EGFP)治疗;qPCR检测各组大鼠视网膜组织中miR-28-3p的表达情况;HE染色观察各组大鼠视网膜内血管分布面积情况;免疫荧光检测CD31的表达;免疫组化检测各组大鼠视网膜组织中Bax和Bcl-2蛋白的表达情况;ELISA检测各组大鼠血清中炎性因子(TNF-α、IL-1β和IL-6)的表达情况;Western blotting检测PI3K、AKT蛋白表达情况。结果:与sham组相比,model组大鼠视网膜组织中miR-28-3p的表达上调(P<0.001),与model组相比,miR-28-3p inhibitor组大鼠视网膜组织中miR-28-3p的表达下调(P<0.001);与sham组相比,model组大鼠的视网膜内血管分布面积和CD31的表达明显增加(均P<0.001),与model组相比,miR-28-3p inhibitor组大鼠视网膜血管分布面积和CD31的表达量均降低(均P<0.01);与model组相比,miR-28-3p inhibitor组中Bax的细胞数量降低(P<0.001),Bcl-2的细胞数量增加(P<0.01);与sham组相比,model组大鼠血清中TNF-α、IL-1β和IL-6含量明显增加(均P<0.001),而miR-28-3p inhibitor组大鼠血清中TNF-α、IL-1β和IL-6含量明显降低(均P<0.01);与model组相比,miR-28-3p inhibitor组大鼠视网膜中PI3K和AKT蛋白含量明显下降(均P<0.01)。结论:miR-28-3p inhibitor能抑制视网膜中血管的生成、减轻炎性反应以及抑制凋亡因子Bax表达,促进凋亡抑制因子Bcl-2表达,并且能抑制PI3K/AKT信号通路的激活,在DR发生发展过程有重要作用。Objective: To investigate the effect of miR-28-3p on of diabetic retinopathy(DR) and retina angiogenesis in rats by PI3K/AKT pathway. Methods: SD rats were divided into sham operation group(sham group), diabetes model group(model group), diabetes + negative control group(miR-NC group) and diabetes+miR-28-3p inhibitor group(miR-28-3p inhibitor group), the diabetic rat model was established by intraperitoneal injection of streptozotocin solution and was treated with adeno-associated virus(AAV-U6-inhibitor-rno-miR-28-3p-CAG-EGFP). qPCR was used to detect the expression of miR-28-3p in retinal tissue of each group. HE staining was used to observe the distribution of blood vessels in the retina of each group. Immunofluorescence was used to detect the expression of CD31. The expression of Bax and Bcl-2 protein in retinal tissue of each group was detected by immunohistochemistry. The expression of inflammatory factors(TNF-α, IL-1β and IL-6) in serum of each group was detected by ELISA. The expression of PI3K and AKT protein was detected by Western blot. Results: Compared with sham group, the expression of miR-28-3p in the retina tissue of rats in the model group was up-regulated(P<0.001). Compared with model group, the expression of miR-28-3p in the retina tissue of rats in the miR-28-3p inhibitor group was down-regulated(P<0.001). Compared with sham group, the distribution area of blood vessels in the retina and the expression of CD31 in the model group increased significantly(both P<0.001). Compared with model group, the distribution area of blood vessels in the retina and the expression of CD31 in the miR-28-3p inhibitor group decreased(both P<0.01). Compared with the model group, the number of Bax cells in the miR-28-3p inhibitor group decreased(P<0.001), but the number of Bcl-2 cells increased(P<0.01). Compared with the sham group, the serum levels of TNF-α, IL-1β and IL-6 in the model group increased significantly(all P<0.001), while the levels of TNF-α, IL-1β and IL-6 in the serum of the miR-28-3p inhibitor

关 键 词：miR-28-3p PI3K/AKT通路 糖尿病性视网膜病变 视网膜血管生成 

分 类 号：R587.2[医药卫生—内分泌] R774.1[医药卫生—内科学]