中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2015年
6期
516-520
,共5页
动脉粥样硬化%微RNAs%α核胞浆转运蛋白类%细胞增殖%细胞运动
動脈粥樣硬化%微RNAs%α覈胞漿轉運蛋白類%細胞增殖%細胞運動
동맥죽양경화%미RNAs%α핵포장전운단백류%세포증식%세포운동
Atherosclerosis%MicroRNAs%alpha Karyopherins%Cell proliferation%Cell movement
目的 观察动脉粥样硬化患者血清中微小RNA-181b(miR-181b)的表达情况,以及miR-181b对人血管内皮细胞核内转移蛋白-α3(Importin-α3)表达和血管平滑肌细胞增殖、迁移的影响,探讨miR-181b在动脉粥样硬化发生发展过程中的作用.方法 选取2012年11月至2013年6月在上海交通大学医学院附属第三人民医院就诊的经外周动脉超声检查诊断为动脉粥样斑块形成的50例患者为动脉粥样硬化组,平均年龄(78.1±8.9)岁;同时入选50名健康成人为对照组,平均年龄(72.5±10.7)岁.采集患者外周血,采用带茎环结构的实时定量PCR方法检测其血清中miR-181b 的表达水平;经Targetscan和Pitar靶基因预测数据库预测核质转运受体Importin-α3为miR-181b的靶基因,体外实验采用Western blot检测miR-181b对人血管内皮细胞Importin-α3表达的影响;采用双荧光素酶报告基因检测证实Importin-α3为miR-181b的直接靶基因;CCK8法和Transwell小室试验检测miR-181b对人血管平滑肌细胞增殖、迁移能力的影响.结果 与正常组比较,动脉粥样硬化组血清中miR-181b的表达水平较低(31.69±0.96比82.28±5.95,P＜0.05);Western-blot和双荧光素酶报告实验证实Importin-α3为miR-181b的直接靶基因;增高miR-181b的表达使得人血管平滑肌细胞的增殖(1.57±0.18比2.66±0.16,P＜0.05)和迁移能力(8.7±1.1比21.4±2.3,P＜0.05)明显降低;抑制miR-181b的表达则获得相反的效果(增殖:2.88±0.09比2.04±0.11,P ＜0.05;迁移:15.2±1.5比8.4±1.3,P＜0.05).结论 动脉粥样硬化患者血清中miR-181b表达水平低于正常人,miR-181b可能通过阻断血管内皮细胞中核因子-κB信号途径和通过抑制血管平滑肌细胞的增殖和迁移起到抗动脉粥样硬化的作用.
目的 觀察動脈粥樣硬化患者血清中微小RNA-181b(miR-181b)的錶達情況,以及miR-181b對人血管內皮細胞覈內轉移蛋白-α3(Importin-α3)錶達和血管平滑肌細胞增殖、遷移的影響,探討miR-181b在動脈粥樣硬化髮生髮展過程中的作用.方法 選取2012年11月至2013年6月在上海交通大學醫學院附屬第三人民醫院就診的經外週動脈超聲檢查診斷為動脈粥樣斑塊形成的50例患者為動脈粥樣硬化組,平均年齡(78.1±8.9)歲;同時入選50名健康成人為對照組,平均年齡(72.5±10.7)歲.採集患者外週血,採用帶莖環結構的實時定量PCR方法檢測其血清中miR-181b 的錶達水平;經Targetscan和Pitar靶基因預測數據庫預測覈質轉運受體Importin-α3為miR-181b的靶基因,體外實驗採用Western blot檢測miR-181b對人血管內皮細胞Importin-α3錶達的影響;採用雙熒光素酶報告基因檢測證實Importin-α3為miR-181b的直接靶基因;CCK8法和Transwell小室試驗檢測miR-181b對人血管平滑肌細胞增殖、遷移能力的影響.結果 與正常組比較,動脈粥樣硬化組血清中miR-181b的錶達水平較低(31.69±0.96比82.28±5.95,P＜0.05);Western-blot和雙熒光素酶報告實驗證實Importin-α3為miR-181b的直接靶基因;增高miR-181b的錶達使得人血管平滑肌細胞的增殖(1.57±0.18比2.66±0.16,P＜0.05)和遷移能力(8.7±1.1比21.4±2.3,P＜0.05)明顯降低;抑製miR-181b的錶達則穫得相反的效果(增殖:2.88±0.09比2.04±0.11,P ＜0.05;遷移:15.2±1.5比8.4±1.3,P＜0.05).結論 動脈粥樣硬化患者血清中miR-181b錶達水平低于正常人,miR-181b可能通過阻斷血管內皮細胞中覈因子-κB信號途徑和通過抑製血管平滑肌細胞的增殖和遷移起到抗動脈粥樣硬化的作用.
목적 관찰동맥죽양경화환자혈청중미소RNA-181b(miR-181b)적표체정황,이급miR-181b대인혈관내피세포핵내전이단백-α3(Importin-α3)표체화혈관평활기세포증식、천이적영향,탐토miR-181b재동맥죽양경화발생발전과정중적작용.방법 선취2012년11월지2013년6월재상해교통대학의학원부속제삼인민의원취진적경외주동맥초성검사진단위동맥죽양반괴형성적50례환자위동맥죽양경화조,평균년령(78.1±8.9)세;동시입선50명건강성인위대조조,평균년령(72.5±10.7)세.채집환자외주혈,채용대경배결구적실시정량PCR방법검측기혈청중miR-181b 적표체수평;경Targetscan화Pitar파기인예측수거고예측핵질전운수체Importin-α3위miR-181b적파기인,체외실험채용Western blot검측miR-181b대인혈관내피세포Importin-α3표체적영향;채용쌍형광소매보고기인검측증실Importin-α3위miR-181b적직접파기인;CCK8법화Transwell소실시험검측miR-181b대인혈관평활기세포증식、천이능력적영향.결과 여정상조비교,동맥죽양경화조혈청중miR-181b적표체수평교저(31.69±0.96비82.28±5.95,P＜0.05);Western-blot화쌍형광소매보고실험증실Importin-α3위miR-181b적직접파기인;증고miR-181b적표체사득인혈관평활기세포적증식(1.57±0.18비2.66±0.16,P＜0.05)화천이능력(8.7±1.1비21.4±2.3,P＜0.05)명현강저;억제miR-181b적표체칙획득상반적효과(증식:2.88±0.09비2.04±0.11,P ＜0.05;천이:15.2±1.5비8.4±1.3,P＜0.05).결론 동맥죽양경화환자혈청중miR-181b표체수평저우정상인,miR-181b가능통과조단혈관내피세포중핵인자-κB신호도경화통과억제혈관평활기세포적증식화천이기도항동맥죽양경화적작용.
Objective To observe the serum expression of miR-181b in atherosclerotic patients and the in vitro effects of miR-181b on vascular smooth muscle cell growth and migration.Methods Fifty patients (mean age:(78.1 ± 8.9) years old) with carotid ultrasound examination evidenced atherosclerotic plaque were enrolled as the atherosclerosis group and 50 healthy (mean age:(72.5 ± 10.7) years old) subjects serve as control group.Stem-loop real time RT-PCR was used to detect the serum expression of miR-181b.Importin-α3 was predicted to be a direct target of miR-181b by Targetscan and Pictar.Western-blot was employed to detect the in vitro effects of miR-181b on the expression of Importin-α3 in endothelial cells.Luciferase reporter assay was employed to testify the prediction.The effects of miR-181b on vascular smooth muscle cell growth,migration abilities were respectively examined by CCK8 assay and Matrigel migration assay.Results Compared with healthy controls,serum expression of miR-181b was significantly downregulated in patients with atherosclerosis (31.69 ± 0.96 vs.82.28 ± 5.95,P ＜ 0.05);Importin-α3 was predicted and proved to be a direct target of miR-181b by Western-blot and luciferase reporter assay.The proliferation and migration of vascular smooth muscle cell were significantly downregulated by forced expression of miR-181 b (1.57 ± 0.18 vs.2.66 ± 0.16,P ＜ 0.05;8.7 ± 1.1 vs.21.4 ± 2.3,P ＜ 0.05),while these effects could be abolished by inhibition of miR-181b(2.88 ±0.09 vs.2.04 ±0.11,P ＜0.05;15.2 ± 1.5 vs.8.4 ± 1.3,P ＜ 0.05).Conclusion The serum miR-181 b level was significantly reduced in patients with atherosclerosis.miR-181b may function as an atherosclerosis suppressor by interupting the NF-κB pathway in endothelial cells and inhibiting the proliferation and migration of vascular smooth muscle cells.
