| miR-302d-3p对高糖环境下人脐静脉内皮细胞增殖、凋亡和迁移的影响及其机制 |
| 投稿时间：2025-01-24 修订日期：2025-02-12 点此下载全文 |
| 引用本文：冯涛,李晶,张鸿源,杜曼·巴戈达提,郑殿宇,马震,潘金强.miR-302d-3p对高糖环境下人脐静脉内皮细胞增殖、凋亡和迁移的影响及其机制[J].医学研究杂志,2025,54(7):77-84 |
| DOI：
10.11969/j.issn.1673-548X.2025.07.015 |
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| 基金项目:新疆维吾尔自治区自然科学基金资助项目(2023D01C147) |
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| 中文摘要:目的 以高糖环境下人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)为研究工具,探讨miR-302d-3p对该细胞的增殖、凋亡和迁移的影响及其可能的机制。方法 将HUVEC随机分为对照组、高糖组、miR-302d-3p mimics NC组(mimics NC组)、miR-302d-3p mimics组、miR-302d-3p inhibitor NC组(inhibitor NC组)以及miR-302d-3p inhibitor组。采用细胞计数试剂盒(cell counting kit-8,CCK-8)检测细胞增殖、流式细胞凋亡检测细胞凋亡率,细胞划痕及Transwell检测细胞迁移,实时荧光定量聚合酶链式反应(polymerase chain reaction,PCR)检测miR-302d-3p和表皮生长因子受体(epidermal growth factor receptor,EGFR)的mRNA表达变化,Western blot法检测ERK磷酸化水平,双荧光素酶报告基因实验,检测miR-302d-3p与EGFR的结合。结果 与对照组比较,高糖组细胞增殖活性、划痕愈合能力、侵袭细胞数降低,细胞凋亡率升高,miR-302d-3p表达水平升高,EGFR、p-ERK表达水平降低。与高糖组比较,miR-302d-3p mimics组增殖活性、划痕愈合能力、侵袭细胞数、p-ERK表达水平降低,凋亡率升高,miR-302d-3p表达水平升高,EGFR表达水平降低,miR-302d-3p inhibitor组增殖活性、划痕愈合能力、侵袭细胞数、p-ERK表达水平升高,凋亡率降低,miR-302d-3p表达水平降低,EGFR表达水平升高。miR-302d-3p mimics+EGFR-WT组荧光强度比值较mimicsNC+EGFR-WT组降低。结论 上调miR-302d-3p表达能抑制高糖环境下HUVECs增殖、迁移及侵袭能力并诱导细胞凋亡,而抑制miR-302d-3p表达的作用则与之相反,其机制可能与miR-302d-3p表达上调可靶向降低EGFR表达水平,下调EGFR/ERK通路活性有关。 |
| 中文关键词:糖尿病足溃疡 miRNA 增殖 转移 机制 |
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| Effect of miR-302d-3p on Proliferation, Apoptosis and Migration of Human Umbilical Vein Endothelial Cells under High Glucose Environment and Its Mechanism. |
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| Abstract:Objective To investigate the effect of miR-302d-3p on the proliferation, apoptosis and migration of human umbilical vein endothelial cell (HUVEC) in high glucose environment and its possible mechanism. Methods HUVEC were randomly divided into control group, high glucose group, miR-302d-3p mimics NC group, miR-302d-3p mimics group, miR-302d-3p inhibitor NC group and miR-302d-3p inhibitor NC group. Cell counting kit-8 (CCK-8) was used to detect cell proliferation, flow cytometry was used to detect the apoptosis rate, cell scratch and Transwell were used to detect cell migration, real-time fluorescent quantitative polymerase chain reaction (PCR) was used to detect the mRNA expression of miR-302d-3p and epidermal growth factor receptor (EGFR), Western blot was used to detect the phosphorylation level of ERK, and dual luciferase reporter gene assay was used to detect the binding of miR-302d-3p to EGFR. Results Compared with the control group, the proliferation activity, wound healing ability and the number of invasive cells in the high glucose group were significantly decreased, the apoptosis rate was significantly increased, the expression level of miR-302d-3p was significantly increased, and the expression levels of EGFR and p-ERK were significantly decreased in the high glucose group. Compared with high glucose group, miR-302d-3p mimics group had lower proliferation activity, wound healing ability, invasive cell number, p-ERK expression level,higher apoptosis rate, higher miR-302d-3p expression level, lower EGFR expression level, while miR-302d-3p inhibitor group had higher proliferation activity, wound healing ability, invasive cell number, p-ERK expression level, lower apoptosis rate, lower miR-302d-3p expression level, and higher EGFR expression level.The fluorescence intensity ratio of miR-302d-3p mimics+egfr-wt group was significantly lower than that of mimicsnc+egfr-wt group, P<0.05. ConclusionUp regulation of miR-302d-3p expression can inhibit the proliferation, migration and invasion of HUVEC under high glucose environment and induce apoptosis, while the effect of inhibiting miR-302d-3p expression is opposite. The mechanism may be related to up regulation of miR-302d-3p expression, which can target to reduce the expression of EGFR and down regulate the activity of EGFR/ERK pathway. |
| keywords:Diabetic foot ulcer miRNA Proliferation Transfer Mechanism |
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