文章摘要
廖清池1 胡艳丽2 周胜华1.MicroRNA-21 介导非对称性二甲基精氨酸诱导的内皮细胞衰老[J].,2011,11(13):2405-2408
MicroRNA-21 介导非对称性二甲基精氨酸诱导的内皮细胞衰老
Effects of Asymmetric Dimethylarginine on Expression of microRNA-21in Endothelial Cells
  
DOI:
中文关键词: MicroRNA-21  ADMA  内皮细胞衰老
英文关键词: MicroRNA-21  Asymmetric dimethylarginine  Endothelial cell senescence
基金项目:国家自然科学基金,资助项目(30871053)
作者单位
廖清池1 胡艳丽2 周胜华1 中南大学湘雅二医院心血管内科 
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中文摘要:
      目的:观察非对称性二甲基精氨酸(ADMA)对内皮细胞中microRNA-21(miR-21)表达的影响,探讨microRNA-21 在ADMA 诱导的内皮细胞衰老中的作用。方法: 人脐静脉内皮细胞(HUVEC)与10 uM 的ADMA 孵育48 小时后收集细胞提取总RNA 及 蛋白,荧光定量实时RT-PCR 检测miR-21 表达,Western blot 检测超氧化物歧化酶2 (SOD2) 表达,衰老相关半乳糖苷酶 (SA-β-gal)染色鉴定衰老的内皮细胞;然后HUVEC 与miR-21 抑制剂转染6 小时后继续与10 uM 的ADMA 孵育48 小时留取细 胞按上述方法检测相关指标。结果:HUVEC 与ADMA 孵育后miR-21 表达量明显增加(P<0.01),同时衰老的内皮细胞数量增多 (P<0.05),而SOD2 表达减少(P<0.01);MiR-21 抑制剂转染HUVEC 后ADMA 诱导的miR-21 表达明显减少,同时衰老的内皮 细胞减少,而SOD2 表达明显增加(所有P<0.01)。结论:ADMA 诱导了HUVEC 中miR-21 表达及细胞衰老,miR-21 介导了 ADMA 诱导的内皮细胞衰老作用,其机制可能与其抑制SOD2 表达有关。
英文摘要:
      Objective: To investigate the effects of asymmetric dimethylarginine (ADMA) on the expression of microRNA-21 (miR-21) in endothelial cells in order to explore the role of miR-21 in ADMA-induced endothelial cell senescence. Methods: After human umbilical vein endothelial cells (HUVECs) were incubated with 10 uM ADMA for 48 hours, the cells were collected for extraction of total RNA and protein. Fluorescence quantitative real-time PCR was applied to determine the expression of miR-21 in HUVECs. Western Blot was used to detect the expression of superoxide dismutase 2 (SOD2) in HUVECs, and senescence associated galactosidase (SA-β-gal) staining was employed to identify the senescent endothelial cells. Then after the transfection of miR-21 inhibitors for 6 hours, HUVEC were incubated with 10 uM ADMA for 48 hours, and then the cells were collected for the determination of miR-21 and SOD2 expression and senescent endothelial cells according to the above-mentioned methods. Results: After HUVECs were incubated with ADMA, the expression of miR-21 was increased significantly (P<0.01) and meanwhile the number of SA-β-gal positive endothelial cells were elevated (P<0.05) , while the SOD2 expression decreased (P<0.01); After the transfection of miR-21 inhibitors the ADMA-induced miR-21 expression in HUVECs was significantly reduced and meanwhile the number of SA-β-gal positive endothelial cells decreased while the SOD2 expression was significantly increased (all P<0.01). Conclusions: ADMA induced the expression of miR-21 in HUVECs and endothelial cell senescence, further miR-21 mediated the endothelial cell senescence induced by ADMA. This mechanism might be related to the inhibition of miR-21 on SOD2 expression.
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