中华老年医学杂志
中華老年醫學雜誌
중화노년의학잡지
Chinese Journal of Geriatrics
2012年
1期
39-43
,共5页
蒋卫东%曾季平%刘玉胜%王欣%秦爱琼%鹿庆华%葛志明
蔣衛東%曾季平%劉玉勝%王訢%秦愛瓊%鹿慶華%葛誌明
장위동%증계평%류옥성%왕흔%진애경%록경화%갈지명
RNA,小分子干扰%动脉粥样硬化%血管生成诱导剂
RNA,小分子榦擾%動脈粥樣硬化%血管生成誘導劑
RNA,소분자간우%동맥죽양경화%혈관생성유도제
RNA,small interfering%Atherosclerosis%Angiogenesis inducing
目的 研究类表皮生长因子域7( Egfl7)在动脉粥样硬化斑块内的表达水平和小分子干扰RNA(siRNA)对内皮细胞株血管生长因子表达的影响. 方法 利用免疫组化和免疫荧光染色检测人髂动脉和小鼠无名动脉粥样硬化斑块内Egfl7的表达水平；并利用以Egfl7为靶标的siRNA,通过脂质体将siRNA转入人脐静脉内皮细胞株(HUVEC),以未转染siRNA细胞和转染无关siRNA细胞为对照,在干扰后0h,12 h,24 h,48 h,反转录聚合酶链反应(PT-PCR)检测Egfl7、血管内皮生长因子(VEGF)、血小板衍生生长因子A、B(PDGF-A、PDGF-B) mRNA水平的改变,免疫印迹法检测Egfl7、VEGF、PDGF、血管细胞黏附分子(VCAM)及细胞间黏附分子(ICAM)蛋白表达水平的变化.结果 人髂动脉和小鼠无名动脉粥样硬化斑块内Egfl7表达水平上调；HUVEC内Egfl7在siRNA干扰后0h、12 h、24 h、48 h时mRNA表达分别为(0.31±0.05)、(0.14±0.02)、(0.09±0.01)、(0.02±0.01),蛋白表达为(0.93±0.08)、(0.71±0.11)、(0.39±0.09)、(0.07±0.01).Egfl7、VEGF、PDGF-A、PDGF-B mRNA及其蛋白表达水平随干扰时间延长均下降或抑制,与siRNA干扰0h比较差异有统计学意义(均P＜0.05). 结论 动脉粥样硬化斑块内Egfl7表达水平提高；siRNA抑制HUVEC Egfl7表达水平的同时,其他血管生长因子和黏附分子表达水平亦下降.
目的 研究類錶皮生長因子域7( Egfl7)在動脈粥樣硬化斑塊內的錶達水平和小分子榦擾RNA(siRNA)對內皮細胞株血管生長因子錶達的影響. 方法 利用免疫組化和免疫熒光染色檢測人髂動脈和小鼠無名動脈粥樣硬化斑塊內Egfl7的錶達水平；併利用以Egfl7為靶標的siRNA,通過脂質體將siRNA轉入人臍靜脈內皮細胞株(HUVEC),以未轉染siRNA細胞和轉染無關siRNA細胞為對照,在榦擾後0h,12 h,24 h,48 h,反轉錄聚閤酶鏈反應(PT-PCR)檢測Egfl7、血管內皮生長因子(VEGF)、血小闆衍生生長因子A、B(PDGF-A、PDGF-B) mRNA水平的改變,免疫印跡法檢測Egfl7、VEGF、PDGF、血管細胞黏附分子(VCAM)及細胞間黏附分子(ICAM)蛋白錶達水平的變化.結果 人髂動脈和小鼠無名動脈粥樣硬化斑塊內Egfl7錶達水平上調；HUVEC內Egfl7在siRNA榦擾後0h、12 h、24 h、48 h時mRNA錶達分彆為(0.31±0.05)、(0.14±0.02)、(0.09±0.01)、(0.02±0.01),蛋白錶達為(0.93±0.08)、(0.71±0.11)、(0.39±0.09)、(0.07±0.01).Egfl7、VEGF、PDGF-A、PDGF-B mRNA及其蛋白錶達水平隨榦擾時間延長均下降或抑製,與siRNA榦擾0h比較差異有統計學意義(均P＜0.05). 結論 動脈粥樣硬化斑塊內Egfl7錶達水平提高；siRNA抑製HUVEC Egfl7錶達水平的同時,其他血管生長因子和黏附分子錶達水平亦下降.
목적 연구류표피생장인자역7( Egfl7)재동맥죽양경화반괴내적표체수평화소분자간우RNA(siRNA)대내피세포주혈관생장인자표체적영향. 방법 이용면역조화화면역형광염색검측인가동맥화소서무명동맥죽양경화반괴내Egfl7적표체수평；병이용이Egfl7위파표적siRNA,통과지질체장siRNA전입인제정맥내피세포주(HUVEC),이미전염siRNA세포화전염무관siRNA세포위대조,재간우후0h,12 h,24 h,48 h,반전록취합매련반응(PT-PCR)검측Egfl7、혈관내피생장인자(VEGF)、혈소판연생생장인자A、B(PDGF-A、PDGF-B) mRNA수평적개변,면역인적법검측Egfl7、VEGF、PDGF、혈관세포점부분자(VCAM)급세포간점부분자(ICAM)단백표체수평적변화.결과 인가동맥화소서무명동맥죽양경화반괴내Egfl7표체수평상조；HUVEC내Egfl7재siRNA간우후0h、12 h、24 h、48 h시mRNA표체분별위(0.31±0.05)、(0.14±0.02)、(0.09±0.01)、(0.02±0.01),단백표체위(0.93±0.08)、(0.71±0.11)、(0.39±0.09)、(0.07±0.01).Egfl7、VEGF、PDGF-A、PDGF-B mRNA급기단백표체수평수간우시간연장균하강혹억제,여siRNA간우0h비교차이유통계학의의(균P＜0.05). 결론 동맥죽양경화반괴내Egfl7표체수평제고；siRNA억제HUVEC Egfl7표체수평적동시,기타혈관생장인자화점부분자표체수평역하강.
Objective To evaluate the expression of epidermal growth factor-like domain 7 (Egfl7) in atherosclerotic plaques and effects of its small interference RNA (siRNA) on angiogenesis gene expression in human endothelial cell line (HUVEC). Methods Egfl7 expression in atherosclerotic plaques were detected in human iliac artery and mouse arteriae using immunohistochemistry and immunofluorescence stainings.The siRNA targeting Egfl7 was transfected into HUVEC by lipofectamine with non-transfected cells and unconcerned siRNA as controls.At 0 h,12 h,24 h and 48 h after intervention,the levels of mRNA and protein of Egf17,vascular endothelial growth factor(VEGF),platelet derived growth factor-A (PDGF-A),platelet derived growth factor-B (PDGF-B),vascular cell adhesion molecule(VCAM) and intercellular adhesion molecule (ICAM)were measured by RT-PCR and Western blotting,respectively. Results The expressions of Egfl7 in human iliac artery and mouse arteriae were increased.The expressions of Egfl7 in HUVEC at the levels of mRNA were[(0.14±0.02),(0.09±0.01),(0.02±0.01)]and the levels of protein[(0.71±0.11),(0.39±0.09),(0.07±0.01)]at 12 h,24 h and 48 h after siRNA intervention,respectively,which were decreased as compared with 0 h intervention [(0.31 ±0.05) and (0.93±0.08) ].Other genes such as VEGF,PDGF-A and PDGF-B were reduced or silenced at the levels of protein and mRNA in HUVEC with siRNA longer interventions(all P＜0.05). Conclusions The expression of Egfl7 in atherosclerotic plaques is increased.The siRNA inhibiting Egfl7 gene expression results in silence of other factors involved in angiogenesis.
