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].现代生物医学进展英文版,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 |
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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) |
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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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