中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
6期
1258-1261
,共4页
血管内超声%动脉粥样硬化%基因治疗%RNA干扰
血管內超聲%動脈粥樣硬化%基因治療%RNA榦擾
혈관내초성%동맥죽양경화%기인치료%RNA간우
Atherosclerosis%Gene therapy%Plaques%RNA interfering
目的 建立猪动脉粥样硬化斑块模型,构建猪靶向小干扰RNA(siRNA)慢病毒载体,定向转染后用血管内超声(IvUS)分析斑块特性变化.方法 60只猪建立动脉粥样硬化模型,间隙连接蛋白37(Cx37) siRNA慢病毒悬液、mock-siRNA、生理盐水分别注入斑块,2个月后用血管内超声检测斑块体积和组织成分,并与干预前比较.同时检测斑块Cx37 mRNA及Cx37蛋白表达,并行组间比较.结果 慢病毒转染2个月后,猪腹主动脉斑块Cx37 mRNA水平在Cx37siRNA组下降到34％,mock-siRNA组下降到61％,生理盐水组下降到94％ (P ＜0.05),其中Cx37siRNA组显著下降.Western blot分析显示Cx37蛋白表达水平在Cx37 siRNA干扰组较其他组低(0.21 ±0.07、0.65±0.06、0.54 ±0.07).IVUS检测,10个月时斑块坏死核心占比率较8个月时[降低(5.26±2.11)％比(7.83±1.03)％,P＜ 0.05].Cx37 siRNA组斑块体积较8个月时减小[(21.03±6.24) mm3比(31.23±10.23) mm3,P＜0.01].而mock-siRNA和生理盐水组,斑块体积较8个月时增大[(38.54±13.56) mm3比(32.12±11.21)mm3,(37.36±14.21) mm3比(30.21±12.02) mm3,P＜0.05].结论 Cx37siRNA安全有效作用于动脉粥样硬化斑块,有效地减少Cx37 mRNA和蛋白表达,并促进斑块稳定及缩小斑块.
目的 建立豬動脈粥樣硬化斑塊模型,構建豬靶嚮小榦擾RNA(siRNA)慢病毒載體,定嚮轉染後用血管內超聲(IvUS)分析斑塊特性變化.方法 60隻豬建立動脈粥樣硬化模型,間隙連接蛋白37(Cx37) siRNA慢病毒懸液、mock-siRNA、生理鹽水分彆註入斑塊,2箇月後用血管內超聲檢測斑塊體積和組織成分,併與榦預前比較.同時檢測斑塊Cx37 mRNA及Cx37蛋白錶達,併行組間比較.結果 慢病毒轉染2箇月後,豬腹主動脈斑塊Cx37 mRNA水平在Cx37siRNA組下降到34％,mock-siRNA組下降到61％,生理鹽水組下降到94％ (P ＜0.05),其中Cx37siRNA組顯著下降.Western blot分析顯示Cx37蛋白錶達水平在Cx37 siRNA榦擾組較其他組低(0.21 ±0.07、0.65±0.06、0.54 ±0.07).IVUS檢測,10箇月時斑塊壞死覈心佔比率較8箇月時[降低(5.26±2.11)％比(7.83±1.03)％,P＜ 0.05].Cx37 siRNA組斑塊體積較8箇月時減小[(21.03±6.24) mm3比(31.23±10.23) mm3,P＜0.01].而mock-siRNA和生理鹽水組,斑塊體積較8箇月時增大[(38.54±13.56) mm3比(32.12±11.21)mm3,(37.36±14.21) mm3比(30.21±12.02) mm3,P＜0.05].結論 Cx37siRNA安全有效作用于動脈粥樣硬化斑塊,有效地減少Cx37 mRNA和蛋白錶達,併促進斑塊穩定及縮小斑塊.
목적 건립저동맥죽양경화반괴모형,구건저파향소간우RNA(siRNA)만병독재체,정향전염후용혈관내초성(IvUS)분석반괴특성변화.방법 60지저건립동맥죽양경화모형,간극련접단백37(Cx37) siRNA만병독현액、mock-siRNA、생리염수분별주입반괴,2개월후용혈관내초성검측반괴체적화조직성분,병여간예전비교.동시검측반괴Cx37 mRNA급Cx37단백표체,병행조간비교.결과 만병독전염2개월후,저복주동맥반괴Cx37 mRNA수평재Cx37siRNA조하강도34％,mock-siRNA조하강도61％,생리염수조하강도94％ (P ＜0.05),기중Cx37siRNA조현저하강.Western blot분석현시Cx37단백표체수평재Cx37 siRNA간우조교기타조저(0.21 ±0.07、0.65±0.06、0.54 ±0.07).IVUS검측,10개월시반괴배사핵심점비솔교8개월시[강저(5.26±2.11)％비(7.83±1.03)％,P＜ 0.05].Cx37 siRNA조반괴체적교8개월시감소[(21.03±6.24) mm3비(31.23±10.23) mm3,P＜0.01].이mock-siRNA화생리염수조,반괴체적교8개월시증대[(38.54±13.56) mm3비(32.12±11.21)mm3,(37.36±14.21) mm3비(30.21±12.02) mm3,P＜0.05].결론 Cx37siRNA안전유효작용우동맥죽양경화반괴,유효지감소Cx37 mRNA화단백표체,병촉진반괴은정급축소반괴.
Objective To construct lentiviruses targeting pig connexin 37 (Cx37) small interfering RNA (siRNA) and establish pig atherosclerosis plaque model,and observe the plaque characteristics by intravenous ultrasound (IVUS) after directional transfection of the lentiviruses into into carotid lesions.Methods The plaque models of 60 pigs fed on high-fat diet were established.The pigs were randomly divided into the saline,mock-siRNA,Cx37 siRNA groups,undergoing transfection of saline,mock-siRNA and Cx37 viral suspension into the abdominal aorta plaques respectively.Two months later,IVUS was used to measure the plaque size and the composition.Simultaneously,the expression of Cx37 mRNA and protein in the plaques was detected.Results Two months after lentivirus transfection,Cx37 mRNA levels in the Cx37 siRNA group,mock-siRNA group and saline group were reduced by 34％,61％,and 94％ respectively (P ＜ 0.05).Western blotting analysis showed that Cx37 protein was lower in Cx37-RNAi group than in other groups (0.21 ±0.07 vs.0.65 ±0.06 vs.0.54 ±0.07).The plaque necrosis on 10-month percentages was decreased in Cx37 siRNA group as compared with 8-month percentages (5.26 ± 2.11 vs.7.83 ± 1.03％,P ＜0.05).In Cx37 siRNA group,plaque volume was decreased relative to the 8-month percentages (21.03 ±6.24 vs.31.23 ± 10.23,P ＜0.01).By contrast,plaque volume was increased relative to their 8-month percentages [(38.54 ± 13.56) mm3 vs.(32.12 ± 11.21) mm3,and (37.36 ± 14.21) mm3 vs.(30.21 ±12.02) mm3,P＜0.05] in mock-siRNA and saline groups.Conclusion By IVUS,Cx37 siRNA exerts an effect on atherosclerotic plaques,and interference of Cx37 reduces the Cx 37 mRNA experssion and decreases the plaques effectively.
