miR-217参与高糖诱导的内皮细胞凋亡的调控 |
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引用本文: | 俞峰,苏陈,张帅,冯金忠,邱一华,周巍,孙俊波,王燕华,吴峰.miR-217参与高糖诱导的内皮细胞凋亡的调控[J].现代生物医学进展,2015,15(13):2429-2431. |
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作者姓名: | 俞峰 苏陈 张帅 冯金忠 邱一华 周巍 孙俊波 王燕华 吴峰 |
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作者单位: | 解放军第九八医院心血管内科 |
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基金项目: | 国家自然科学基金项目(81400207) |
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摘 要: | 目的:研究miR-217对高糖诱导的内皮刺激内皮细胞凋亡的作用。方法:培养人冠状动脉内皮细胞,用含D-葡萄糖(30mmol/L)的培养液刺激:(1)利用实时定量PCR检测内皮细胞相关微小RNA(miR-217、miR-137、miR-29c、miR-218、miR-451、miR-328、miR-517c和miR-216a等)的表达变化;(2)利用慢病毒感染技术干预内皮细胞miR-217水平,利用流式细胞术检测细胞凋亡水平;(3)生物信息学预测、双荧光素酶报告基因验证以及蛋白免疫印迹法(western blot)确定miR-217的靶基因。结果:(1)实时定量PCR检测发现高糖刺激内皮细胞后,miR-217、miR-137、miR-29c、miR-218等的表达上调(P0.01),miR-451、miR-328、miR-517c和miR-216a的表达下调(P0.01),其中miR-217比对照细胞升高了5.67倍;(2)慢病毒感染内皮细胞后再经高糖刺激,流式细胞术检测发现过表达miR-217的内皮细胞凋亡水平有显著提高;(3)生物信息学分析发现SIRT1基因的3'非翻译区上存在一个miR-217的结合位点,双荧光素酶报告基因和western blot结果均证明SIRT1是miR-217的靶基因。结论:miR-217可能通过抑制SIRT1的表达参与高糖诱导的内皮细胞凋亡的调控。
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关 键 词: | miR-217 内皮细胞 高糖 凋亡 |
MiR-217 may Involve in Apoptosis Regulation of Endothelial Cell Injury
Induced by High-glucose |
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Abstract: | Objective:To study the effect of miR-217 on apoptosis of endothelial cell induced by high-glucose.Methods:Human
coronary artery endothelial cells were culture by medium with 30 mmol/L D-glucose, and related microRNAs (miR-217, miR-137,
miR-29c, miR-218, miR-451, miR-328, miR-517c and miR-216a) were detected by real-time PCR. The cell apoptosis level of endothelial
cells overexpressed miR-217 by using lentivirus infection technology was investigated by flow cytometry. Then, the targeted gene of
miR-217 was identified by using bioinformatics, dual-luciferase assay and western blot.Results:Real-time PCR assay showed that
miR-217, miR-137, miR-29c and miR-218 were significantly increased (P<0.01) after high-glucose stimulating, while miR-451,
miR-328, miR-517c and miR-216a were significantly decreased (P<0.01). Especially, the miR-217 level was increased 5.67-fold than that
of control cell. After overexpressing miR-217 by using lentivirus infection, these cells apoptosis level was obviously increased. Bioinformatics
prediction found a binding-site in 3'' untranslated region of SIRT1, this site was a targeted bind site of miR-217 confirmed by dualluciferase
assay and western blot.Conclusion:MiR-217 may involve in apoptosis regulation of high-glucose induced endothelial cell injury
by inhibiting expression of SIRT1. |
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Keywords: | MiR-217 Endothelial cell injury High-glucose Apoptosis |
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