中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2014年
8期
1415-1420
,共6页
张文宇%王辉%李玉媚%陈伟%胡楠%欧和生
張文宇%王輝%李玉媚%陳偉%鬍楠%歐和生
장문우%왕휘%리옥미%진위%호남%구화생
miRNA-24%细胞增殖%一氧化氮合酶%一氧化氮%Sp1转录因子
miRNA-24%細胞增殖%一氧化氮閤酶%一氧化氮%Sp1轉錄因子
miRNA-24%세포증식%일양화담합매%일양화담%Sp1전록인자
miRNA-24%Cell proliferation%Nitric oxide synthase%Nitric oxide%Sp1 transcription factor
目的:为进一步探讨miRNA-24是否参与内皮型一氧化氮合酶(eNOS)表达的调控及其对血管内皮细胞增殖的影响,本研究构建miRNA-24高表达质粒,并用脂质体将其导入人脐静脉内皮细胞( HUVECs ),观察miRNA-24对eNOS表达及HUVECs增殖的影响。方法:用四甲基偶氮唑蓝( MTT)法检测细胞增殖情况,RT-PCR、免疫组织化学和Western blotting检测eNOS与Sp1转录因子mRNA和蛋白表达水平。结果:与对照组比较, miR-NA-24高表达组细胞增殖减慢41.97%(0.47±0.04 vs 0.81±0.03, P<0.01),eNOS mRNA降低44.8%(0.48±0.01 vs 0.87±0.03,P<0.05),蛋白表达减少71.92%(0.16±0.06 vs 0.57±0.08,P<0.05);同时Sp1 mRNA降低53.00%(0.45±0.02 vs 0.93±0.01,P<0.05),其蛋白质表达量也相应减少62.31%(0.13±0.07 vs 0.31±0.09, P<0.05)。 miRNA-24抑制组中,上述指标比对照组降低,但比miRNA-24高表达组明显升高。结论:miRNA-24高表达明显抑制HUVECs的增殖及eNOS的表达;Sp1可能是参与miRNA-24调控eNOS表达的重要因素之一。
目的:為進一步探討miRNA-24是否參與內皮型一氧化氮閤酶(eNOS)錶達的調控及其對血管內皮細胞增殖的影響,本研究構建miRNA-24高錶達質粒,併用脂質體將其導入人臍靜脈內皮細胞( HUVECs ),觀察miRNA-24對eNOS錶達及HUVECs增殖的影響。方法:用四甲基偶氮唑藍( MTT)法檢測細胞增殖情況,RT-PCR、免疫組織化學和Western blotting檢測eNOS與Sp1轉錄因子mRNA和蛋白錶達水平。結果:與對照組比較, miR-NA-24高錶達組細胞增殖減慢41.97%(0.47±0.04 vs 0.81±0.03, P<0.01),eNOS mRNA降低44.8%(0.48±0.01 vs 0.87±0.03,P<0.05),蛋白錶達減少71.92%(0.16±0.06 vs 0.57±0.08,P<0.05);同時Sp1 mRNA降低53.00%(0.45±0.02 vs 0.93±0.01,P<0.05),其蛋白質錶達量也相應減少62.31%(0.13±0.07 vs 0.31±0.09, P<0.05)。 miRNA-24抑製組中,上述指標比對照組降低,但比miRNA-24高錶達組明顯升高。結論:miRNA-24高錶達明顯抑製HUVECs的增殖及eNOS的錶達;Sp1可能是參與miRNA-24調控eNOS錶達的重要因素之一。
목적:위진일보탐토miRNA-24시부삼여내피형일양화담합매(eNOS)표체적조공급기대혈관내피세포증식적영향,본연구구건miRNA-24고표체질립,병용지질체장기도입인제정맥내피세포( HUVECs ),관찰miRNA-24대eNOS표체급HUVECs증식적영향。방법:용사갑기우담서람( MTT)법검측세포증식정황,RT-PCR、면역조직화학화Western blotting검측eNOS여Sp1전록인자mRNA화단백표체수평。결과:여대조조비교, miR-NA-24고표체조세포증식감만41.97%(0.47±0.04 vs 0.81±0.03, P<0.01),eNOS mRNA강저44.8%(0.48±0.01 vs 0.87±0.03,P<0.05),단백표체감소71.92%(0.16±0.06 vs 0.57±0.08,P<0.05);동시Sp1 mRNA강저53.00%(0.45±0.02 vs 0.93±0.01,P<0.05),기단백질표체량야상응감소62.31%(0.13±0.07 vs 0.31±0.09, P<0.05)。 miRNA-24억제조중,상술지표비대조조강저,단비miRNA-24고표체조명현승고。결론:miRNA-24고표체명현억제HUVECs적증식급eNOS적표체;Sp1가능시삼여miRNA-24조공eNOS표체적중요인소지일。
[ABSTRACT]AIM:ToinvestigatewhethermiRNA-24isinvolvedintheregulationofendothelialnitricoxide synthase ( eNOS ) expression and vascular endothelial cell proliferation .METHODS: A plasmid that highly expressed miRNA-24 was constructed, and was transfected into the human umbilical vein endothelial cells (HUVECs) by liposome. The cell proliferation was detected by MTT assay .The expression of eNOS and Sp 1 at mRNA and protein levels was exa-mined by real-time PCR, immunohistochemistry and Western blotting .RESULTS:Compared with control group , the pro-liferation of endothelial cells in miRNA-24 group was significantly decreased by 41.97 % (0.47 ±0.04 vs 0.81 ±0.03, P<0.01), and the expression of eNOS at mRNA and protein levels was decreased by 44.8% (0.48 ±0.01 vs 0.87 ± 0.03, P<0.05) and 71.92%(0.16 ±0.06 vs 0.57 ±0.08, P<0.05), respectively.Meanwhile, the mRNA and pro-tein levels of Sp1 were significantly decreased by 53.00% (0.45 ±0.02 vs 0.93 ±0.01, P<0.05) and by 62.31%(0.13 ±0.07 vs 0.31 ±0.09, P<0.05), respectively.In miRNA-24 inhibitor group, the above indexes were decreased compared with control group , but significantly increased compared with miRNA-24 group.CONCLUSION: miRNA-24 significantly inhibits the proliferation of HUVECs and the eNOS expression .Sp1 possibly acts as one of the important factors in the regulation of eNOS expression by miRNA-24.