He Jianhong, Wei Shuzhen, Ye Lingzhi, Liao Rifang, Zeng Zhuanping
Department of Epidemiology and Health Statistics, School of Public Health, Guangdong Pharmaceutical University, Guangzhou 510006, China.
Department of Pharmacy, Sun Yat-Sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China.
Gene. 2025 Sep 5;963:149599. doi: 10.1016/j.gene.2025.149599. Epub 2025 May 30.
Whether miR-29a-5p is associated with Diabetic Retinopathy (DR) is still a subject of ongoing discussion. The current research examines the involvement of miR-29a-5p in regulating apoptosis, oxidative stress, and inflammation of retinal ganglion cells (RGCs) generated by High Glucose (HG). Additionally, we are interested in analyzing the contribution of miR-29a-5p in DRdeveloping.
We obtained peripheral blood samples from 7 people with DR and 14 individuals without DR. Subsequently, we conducted biochemical indices analyses and qRT-PCR.We randomly divided RGCs into low glucose groups, HG groups, HG + inhibitor negative control groups, and HG + miR-29a-5p inhibitor groups. SIRT3 siRNA was transfected into RGCs through lipofectamine 3000 reagent.Cell vitality was detected by MTT; qRT-PCR was applied to identify miR-29a-5p expression; Detectionof ROS, SOD, and MDAlevels was quantified using the DCFH-DA, WST-1, and colorimetric methods, respectively; IL-6 and TNF-αcontents were analyzed utilizing ELISA; Dual-luciferase gene reporter experiment was used to examine if SIRT3 is the specific target gene of miR-29a-5p; Flow cytometryevaluatedapoptosisin RGCs; The technique of Western blotting identified the presence of caspase-3 proteins.
The expression of miR-29a-5p was markedly elevated in individuals with DR, and it had positive correlations with levels of Total Cholesterol (TC) and Fasting Blood Glucose (FBG).High glucose significantly induced RGC apoptosis and upregulated the miR-29a-5p gene. Transfection of miR-29a-5p inhibitor protected RGCs against HG-induced oxidative injury, inflammation, and apoptosis. Furthermore, the dual-luciferase reporter experiment provided confirmation that SIRT3 was a target gene of miR-29a-5p, as it negatively regulated SIRT3 expression. Notably, SIRT3 knockdown abolished the protection of miR-29a-5p inhibition on RGCs.
Suppression of the miR-29a-5p gene safeguards RGCs against harm caused by HG via boosting SIRT3 signaling, which might provide a new prevention and treatment strategy for DR.
miR-29a-5p是否与糖尿病视网膜病变（DR）相关仍是一个正在讨论的话题。当前研究探讨了miR-29a-5p在调节高糖（HG）诱导的视网膜神经节细胞（RGCs）凋亡、氧化应激和炎症中的作用。此外，我们还对分析miR-29a-5p在DR发生发展中的作用感兴趣。
我们从7例DR患者和14例非DR个体中获取外周血样本。随后，我们进行了生化指标分析和qRT-PCR。我们将RGCs随机分为低糖组、HG组、HG + 抑制剂阴性对照组和HG + miR-29a-5p抑制剂组。通过脂质体3000试剂将SIRT3 siRNA转染到RGCs中。用MTT检测细胞活力；应用qRT-PCR鉴定miR-29a-5p表达；分别使用DCFH-DA、WST-1和比色法对ROS、SOD和MDA水平进行定量检测；利用ELISA分析IL-6和TNF-α含量；采用双荧光素酶基因报告实验检测SIRT3是否为miR-29a-5p的特异性靶基因；流式细胞术评估RGCs中的凋亡情况；蛋白质印迹技术鉴定caspase-3蛋白的存在。
DR患者中miR-29a-5p的表达显著升高，且与总胆固醇（TC）和空腹血糖（FBG）水平呈正相关。高糖显著诱导RGCs凋亡并上调miR-29a-5p基因。转染miR-29a-5p抑制剂可保护RGCs免受HG诱导的氧化损伤和炎症及凋亡。此外，双荧光素酶报告实验证实SIRT3是miR-29a-5p的靶基因，因为它负向调节SIRT3表达。值得注意的是，敲低SIRT3消除了miR-29a-5p抑制对RGCs的保护作用。
抑制miR-29a-5p基因可通过增强SIRT3信号通路保护RGCs免受HG的损害，这可能为DR提供一种新的防治策略。
