高糖作用下人脐静脉内皮细胞microrna差异表达谱分析-内科学(内分泌与代谢)专业论文.docxVIP

PAGE PAGE 4 高糖作用下人脐静脉内皮细胞 microRNA 差异表达谱分析 中文摘要 目的: 研究高糖作用下的人脐静脉内皮细胞和正常培养的人脐静脉内皮细 胞 microRNA 的差异表达谱。 方法: 利用 microRNA 芯片技术（此芯片包含人类所有成熟 microRNA 及 microRNA 前体序列）筛选出高糖培养基（正常培养基添加葡萄糖至最终糖浓度 为 30.5 mmol/L）培养的人脐静脉内皮细胞相对于正常培养基（M199 添加 10%胎 牛血清, 糖浓度为 5.5 mmol/L）培养的人脐静脉内皮细胞差异表达的 microRNA。 然后采用实时荧光定量 PCR 技术验证 microRNA 芯片结果的可靠性。 结果: 通过 microRNA 芯片筛选发现有 95 个 microRNAs 在高糖组人脐静脉内 皮细胞和正常组人脐静脉内皮细胞之间差异表达，其中高糖组相对于正常组有 61 个 microRNAs 高表达，34 个 microRNAs 低表达。利用实时荧光定量 PCR 验证 结果证明 microRNA 芯片结果是可靠的。 结论: 我们研究发现在高糖作用下人脐静脉内皮细胞中 microRNA 的表达谱 会发生改变，为深入研究糖尿病及其血管并发症的发生机制提供了有意义的线索 同时也为预防、诊断、治疗糖尿病及其血管并发症提供了有用的数据。然而，要 全面了解这些差异表达的 microRNAs 在糖尿病及其血管并发症发生机制中的具 体功能作用还需要我们做进一步的研究。 关键词: microRNA 糖尿病 糖尿病血管并发症 高糖 人脐静脉内皮细胞 Differential expression profiles of microRNA in human umbilical vein endothelial cells induced by high glucose Abstract Objective: We aim to investigate the differential expression of miRNAs in human umbilical vein endothelial cells (HUVECs) cultured with high glucose (HG) medium compared with normal medium. Methods: We analyzed the miRNA expression pro?les in HUVECs incubated under HG condition [normal glucose (NG) medium supplemented with glucose to a ?nal concentration of 30.5 mmol/L] and normal condition (medium 199 plus 10% FBS), using a mammalian miRNA microarray containing whole human mature and precursor miRNA sequences. The array results were confirmed by quantitative real-time PCR (qRT-PCR). Results: A total of 95 miRNAs were detected differentially expressed between the HG group and the normal group by the miRNA microarray chip, while 61 miRNAs up-regulated and 34 miRNAs down-regulated in high glucose cultured cells. The qRT-PCR data were consistent with miRNA microarray results. Conclusion: In the present study, we found that many miRNA expressions were altered in HUVECs under high glucose condition. Our results may provide a valuable clue for the pathogenesis of diabetes and its related vascular complications and lead to finding novel methods to prevent, diagnose, and tr