实用临床医药杂志2025，Vol.29Issue(11)：14-19,25,7.DOI:10.7619/jcmp.20246117
微小RNA-146a-5p对高糖诱导的肾小管上皮细胞损伤的影响及机制研究
Effect and mechanism of microRNA-146a-5p on high glucose-induced injury of renal tubular epithelial cells
摘要
Abstract
Objective To investigate the effect of microRNA-146a-5p(miR-146a-5p)on high glucose-induced injury of renal tubular epithelial cells by regulating high mobility group A2(HMGA2).Methods Human renal tubular epithelial cells(HK-2)were cultured in vitro and di-vided into different groups according to various treatment methods:normal control group(Con group),high glucose group(HG group),HG+miR-146a-5p group,HG+si-HMGA2 group,HG+miR-146a-5p+pcDNA-HMGA2 group,miR-NC group,miR-146a-5p group,anti-miR-NC group,and anti-miR-146a-5p group.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression levels of miR-146a-5p and HMGA2 mRNA in each group.A dual-luciferase reporter assay was employed to verify the targeting relationship between miR-146a-5p and HMGA2.Western blot was utilized to detect the expression levels of cleaved-caspase3,cleaved-caspase9,and HMGA2 protein in each group.Flow cytometry was applied to determine the apoptosis rate of cells in each group.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of inflam-matory factors[interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interleukin-8(IL-8)]in each group.The levels of oxidative stress indicators[catalase(CAT),superoxide dismutase(SOD),malondialdehyde(MDA)]in each group were also measured.Results A nucleotide se-quence complementary to miR-146a-5p was found in the 3'-untranslated region(3'-UTR)of HMGA2.The expression level of HMGA2 protein in the miR-146a-5p group was lower than that in the miR-NC group,while was higher in the anti-miR-146a-5p group than that in the anti-miR-NC group(P＜0.05).The total apoptosis rate,early apoptosis rate,late apoptosis rate,and the levels of cleaved-caspase3 protein,cleaved-caspase9 protein,HMGA2 mRNA,HMGA2 protein,TNF-α,IL-6,IL-8,and MDA in the HG group were higher than those in the Con group,whereas the levels of miR-146a-5p,CAT,and SOD were lower(P＜0.05).In the HG+miR-146a-5p group,these indicators showed opposite trends compared to the HG group(P＜0.05).Similarly,in the HG+si-HMGA2 group,the total apoptosis rate,early apoptosis rate,late apoptosis rate,and the levels of cleaved-caspase3 protein,cleaved-caspase9 protein,HMGA2 mRNA,HMGA2 protein,TNF-α,IL-6,IL-8,and MDA were lower than those in the HG group,while the levels of CAT and SOD were higher(P＜0.05).In the HG+miR-146a-5p+pcDNA-HMGA2 group,the trends of these indicators were consistent with those between the HG group and the HG+miR-146a-5p group,showing statistically significant differences when compared to the HG+miR-146a-5p group(P＜0.05).Conclusion MiR-146a-5p may play a regulatory role in high glucose-induced injury of re-nal tubular epithelial cells by targeting HMGA2.关键词
微小RNA-146a-5p/高迁移率族蛋白A2/肾小管上皮细胞/高糖/细胞凋亡/氧化应激/炎症反应/糖尿病肾病Key words
microRNA-146a-5p/high mobility group A2/renal tubular epithelial cells/high glucose/cell apoptosis/oxidative stress/inflammatory response/diabetic nephropathy分类
医药卫生引用本文复制引用
王晨鸽,杜海梅,尹东升..微小RNA-146a-5p对高糖诱导的肾小管上皮细胞损伤的影响及机制研究[J].实用临床医药杂志,2025,29(11):14-19,25,7.基金项目
陕西省自然科学基金项目(2020FR26003) （2020FR26003）
