中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2013年
5期
406-410
,共5页
许程洁%王京伟%宋健%武军驻%宋婷婷
許程潔%王京偉%宋健%武軍駐%宋婷婷
허정길%왕경위%송건%무군주%송정정
动脉粥样硬化%内皮细胞%α防御素%活性氧%脂蛋白类,LDL
動脈粥樣硬化%內皮細胞%α防禦素%活性氧%脂蛋白類,LDL
동맥죽양경화%내피세포%α방어소%활성양%지단백류,LDL
Atherosclerosis%Endothelial cells%Alpha-defensins%Reactive oxygen species%Lipoproteins,LDL
目的 探讨人α-防御素(human α-defensin,HNP)-1对人内皮细胞氧化低密度脂蛋白(low density lipoprotein,LDL)的影响及其可能机制.方法 实验设HNP-1转染组(HNP-1转染的内皮细胞ECV304),siRNA干扰组(siRNA干扰抑制HNP-1表达的ECV304)和HNP-1刺激组(10 μg/mlHNP-1刺激的ECV304),并分别设有正常ECV304作为对照组,均经LDL处理3h后检测脂质氧化产物丙二醛(M DA)、蛋白氧化产物PCO(protein carbonyl,PGO)的生成量.分别采用流式细胞仪、荧光显微镜检测经LDL、脂多糖(LPS)、HNP-1预处理的ECV304(分别为LDL组、LPS组、HNP-1刺激组)和过表达HNP-1的ECV304(HNP-1转染组)中氧自由基的生成量,另设正常ECV304作为对照组.结果 (1) MDA和PCO生成量检测结果:HNP-1刺激组、HNP-1转染组MDA生成量均明显高于其相应的对照组[分别为(14.49±1.10)比(9.47±1.18) nmol/mg蛋白和(4.21 ±0.03)比(3.15±0.02)nmol/mg蛋白,P均＜0.05].siRNA干扰组MDA生成量则明显低于其对照组[(3.76±0.48)比(4.54 ±0.28) nmol/mg蛋白,P＜0.05].HNP-1转染组PCO生成量明显高于对照组和siRNA干扰组(P均＜0.C5),且siRNA干扰组较对照组低,但差异无统计学意义.(2)氧自由基水平检测结果:HNP-1刺激组和HNP-1转染组ECV304内氧自由基水平均较LDL组、LPS组及对照组高(P均＜0.05).结论 HNP-1可增强人内皮细胞ECV304氧化LDL的能力,机制可能与提高氧自由基的表达水平有关.
目的 探討人α-防禦素(human α-defensin,HNP)-1對人內皮細胞氧化低密度脂蛋白(low density lipoprotein,LDL)的影響及其可能機製.方法 實驗設HNP-1轉染組(HNP-1轉染的內皮細胞ECV304),siRNA榦擾組(siRNA榦擾抑製HNP-1錶達的ECV304)和HNP-1刺激組(10 μg/mlHNP-1刺激的ECV304),併分彆設有正常ECV304作為對照組,均經LDL處理3h後檢測脂質氧化產物丙二醛(M DA)、蛋白氧化產物PCO(protein carbonyl,PGO)的生成量.分彆採用流式細胞儀、熒光顯微鏡檢測經LDL、脂多糖(LPS)、HNP-1預處理的ECV304(分彆為LDL組、LPS組、HNP-1刺激組)和過錶達HNP-1的ECV304(HNP-1轉染組)中氧自由基的生成量,另設正常ECV304作為對照組.結果 (1) MDA和PCO生成量檢測結果:HNP-1刺激組、HNP-1轉染組MDA生成量均明顯高于其相應的對照組[分彆為(14.49±1.10)比(9.47±1.18) nmol/mg蛋白和(4.21 ±0.03)比(3.15±0.02)nmol/mg蛋白,P均＜0.05].siRNA榦擾組MDA生成量則明顯低于其對照組[(3.76±0.48)比(4.54 ±0.28) nmol/mg蛋白,P＜0.05].HNP-1轉染組PCO生成量明顯高于對照組和siRNA榦擾組(P均＜0.C5),且siRNA榦擾組較對照組低,但差異無統計學意義.(2)氧自由基水平檢測結果:HNP-1刺激組和HNP-1轉染組ECV304內氧自由基水平均較LDL組、LPS組及對照組高(P均＜0.05).結論 HNP-1可增彊人內皮細胞ECV304氧化LDL的能力,機製可能與提高氧自由基的錶達水平有關.
목적 탐토인α-방어소(human α-defensin,HNP)-1대인내피세포양화저밀도지단백(low density lipoprotein,LDL)적영향급기가능궤제.방법 실험설HNP-1전염조(HNP-1전염적내피세포ECV304),siRNA간우조(siRNA간우억제HNP-1표체적ECV304)화HNP-1자격조(10 μg/mlHNP-1자격적ECV304),병분별설유정상ECV304작위대조조,균경LDL처리3h후검측지질양화산물병이철(M DA)、단백양화산물PCO(protein carbonyl,PGO)적생성량.분별채용류식세포의、형광현미경검측경LDL、지다당(LPS)、HNP-1예처리적ECV304(분별위LDL조、LPS조、HNP-1자격조)화과표체HNP-1적ECV304(HNP-1전염조)중양자유기적생성량,령설정상ECV304작위대조조.결과 (1) MDA화PCO생성량검측결과:HNP-1자격조、HNP-1전염조MDA생성량균명현고우기상응적대조조[분별위(14.49±1.10)비(9.47±1.18) nmol/mg단백화(4.21 ±0.03)비(3.15±0.02)nmol/mg단백,P균＜0.05].siRNA간우조MDA생성량칙명현저우기대조조[(3.76±0.48)비(4.54 ±0.28) nmol/mg단백,P＜0.05].HNP-1전염조PCO생성량명현고우대조조화siRNA간우조(P균＜0.C5),차siRNA간우조교대조조저,단차이무통계학의의.(2)양자유기수평검측결과:HNP-1자격조화HNP-1전염조ECV304내양자유기수평균교LDL조、LPS조급대조조고(P균＜0.05).결론 HNP-1가증강인내피세포ECV304양화LDL적능력,궤제가능여제고양자유기적표체수평유관.
Objective To explore the role and potential mechanism of human α-defensin 1 (HNP-1) on low-density lipoprotein (LDL) oxidation ability of human endothenial cells (EVC304).Methods Post incubation with LDL for 3 h,the malondiadehyde (MDA) and protein carbonyl (PCO) were detected in untreated ECV304 (control) and in HNP-1 transfected ECV304 in the presence and absence of siRNA against HNP-1.Flow cytometry and fluorescence microscopy were used to detect the generation of oxygen free radical in the ECV304 which have been pretreated by LDL,LPS and HNP-1,respectively.Result Compared with control group,MDA level was significantly increased in HNP-1 transfected[(4.21 ± 0.03) vs.(3.15 ± 0.02) nmol/mg · pro] or in HNP-1stimulated ECV304 cells [(14.49 ± 1.10) vs.(9.47 ± 1.18) nmol/mg · pro],which could be significantly downregulated by siRNA [(3.76 ± 0.48) vs.(4.54 ± 0.28) nmol/mg · pro,all P ＜ 0.05].PCO was also significantly increased in HNP-1 transfected ECV304 cells.The levels of free radical were significantly increased in HNP-1 transfected or HNP-1 stimulated ECV304 cells.Conclusion HNP-1 can enhance the LDL oxidation ability of human endothelial cells via promoting the generation of free radicals.
