抑制磷酸二酯酶5的短发夹RNA重组腺病毒载体可改善小鼠心肌梗死后心力衰竭
억제린산이지매5적단발협RNA중조선병독재체가개선소서심기경사후심력쇠갈
Adenoviral short hairpin RNA targeting phosphodiesterase 5 attenuates cardiac remodeling and cardiac dysfunction following myocardial infarction in mice
  • 万方数据
    中华心血管病杂志 중화심혈관병잡지
    Chinese Journal of Cardiology
    2014年 4期 321-326 ,共6页

    张健%金哲%李龙虎%刚丽%于勤%王美兰%宋爱琳%洪炳哲

    장건%금철%리룡호%강려%우근%왕미란%송애림%홍병철

    心肌梗死%RNA%磷酸二酯酶5 心肌梗死%RNA%燐痠二酯酶5
    심기경사%RNA%린산이지매5
    Myocardial infarction%RNA%Phosphodiesterase 5
    目的 探讨抑制磷酸二酯酶5(PDE5)的短发夹RNA重组腺病毒载体(PDE5 shRNA)对小鼠心肌梗死后心室重构和心力衰竭的影响.方法 雄性10周龄C57BL/6J小鼠,随机分为假手术组(仅穿线不结扎左冠状动脉前降支,n=6)、PDE5 shRNA组(结扎左冠状动脉前降支+心肌注射PDE5shRNA,n=12)、普通腺病毒组(结扎左冠状动脉前降支+心肌注射普通腺病毒,n=15)和DMEM组(结扎左冠状动脉前降支+心肌注射DMEM,n=8).4周后,统计各组小鼠存活情况,通过心脏超声检查和组织学染色的方法分别检测各组小鼠左心室射血分数(LVEF)、左心室短轴缩短率(LVFS)、左心室舒张末期内径(LVEDD)、左心室收缩末期内径(LVESD)、心肌梗死面积和心肌纤维化面积,以评价PDE5shRNA对小鼠心功能及心室重构的影响.同时采用免疫组织化学、ELISA、Western blot等方法,检测血管密度以及PDE5、环磷酸鸟苷(cGMP)和蛋白激酶G(PKG)等蛋白的表达情况.结果 (1)小鼠存活情况:4周后,假手术组小鼠全部存活(100％),DMEM组死亡3只(37％),PDE5 shRNA组小鼠死亡1只(8％)与普通腺病毒组死亡5只(33％)比较差异无统计学意义(P=0.1 1).(2)心功能和心室重构指标的检测结果:PDE5shRNA组小鼠心肌梗死面积显著小于普通腺病毒组和DMEM组[(25.4±2.9)％比(42.0±3.2)％和(43.4±2.6)％,P均＜0.05],LVFS显著高于普通腺病毒组和DMEM组[(21.1l±3.72)％比(14.22±2.91)％和(14.22±2.91)％,P均＜0.05],低于假手术组(41.3±1.8)％(P＜0.05),LVEF显著高于普通腺病毒组和DMEM组[(48.23±7.06)％比(34.59±6.23)％和(38.1±2.8)％,P均＜0.05],低于假手术组(77.7±3.1)％(P＜0.05),LVEDD显著小于普通腺病毒组和DMEM组[(4.88 ±0.36)mm比(5.72±0.62)mm和(5.49 ±0.37)mm,P均＜0.05],大于假手术组(3.26 ±0.29) mm(P ＜0.05),LVESD显著小于普通腺病毒组和DMEM组[(3.87±0.45) mm比(4.91±0.62) mm和(4.63±0.37) mm,P均＜0.05],大于假手术组(1.93 ±0.25) mm(P ＜0.05).(3)血管密度检测结果:PDE5 shRNA组小鼠血管密度显著高于普通腺病毒组,心肌梗死区为(14.3±2.0)个比(6.6±1.2)个(P＜0.05)、梗死周边为(23.6±2.1)个比(13.7±2.4)个(P＜0.05).(4) PDE5蛋白表达水平检测结果:PDE5 shRNA组显著低于普通腺病毒组,0.448 ±0.105比0.993±0.057(P ＜0.05).(5) cGMP和PKG蛋白表达的检测结果:PDE5 shRNA组两种蛋白的表达水平均显著高于对照组(P均＜0.05).结论 PDE5 shRNA可能通过上调cGMP和PKG表达减少小鼠心肌梗死面积和抑制心肌纤维化,从而抑制心室重构、改善心功能.
    목적 탐토억제린산이지매5(PDE5)적단발협RNA중조선병독재체(PDE5 shRNA)대소서심기경사후심실중구화심력쇠갈적영향.방법 웅성10주령C57BL/6J소서,수궤분위가수술조(부천선불결찰좌관상동맥전강지,n=6)、PDE5 shRNA조(결찰좌관상동맥전강지+심기주사PDE5shRNA,n=12)、보통선병독조(결찰좌관상동맥전강지+심기주사보통선병독,n=15)화DMEM조(결찰좌관상동맥전강지+심기주사DMEM,n=8).4주후,통계각조소서존활정황,통과심장초성검사화조직학염색적방법분별검측각조소서좌심실사혈분수(LVEF)、좌심실단축축단솔(LVFS)、좌심실서장말기내경(LVEDD)、좌심실수축말기내경(LVESD)、심기경사면적화심기섬유화면적,이평개PDE5shRNA대소서심공능급심실중구적영향.동시채용면역조직화학、ELISA、Western blot등방법,검측혈관밀도이급PDE5、배린산조감(cGMP)화단백격매G(PKG)등단백적표체정황.결과 (1)소서존활정황:4주후,가수술조소서전부존활(100％),DMEM조사망3지(37％),PDE5 shRNA조소서사망1지(8％)여보통선병독조사망5지(33％)비교차이무통계학의의(P=0.1 1).(2)심공능화심실중구지표적검측결과:PDE5shRNA조소서심기경사면적현저소우보통선병독조화DMEM조[(25.4±2.9)％비(42.0±3.2)％화(43.4±2.6)％,P균＜0.05],LVFS현저고우보통선병독조화DMEM조[(21.1l±3.72)％비(14.22±2.91)％화(14.22±2.91)％,P균＜0.05],저우가수술조(41.3±1.8)％(P＜0.05),LVEF현저고우보통선병독조화DMEM조[(48.23±7.06)％비(34.59±6.23)％화(38.1±2.8)％,P균＜0.05],저우가수술조(77.7±3.1)％(P＜0.05),LVEDD현저소우보통선병독조화DMEM조[(4.88 ±0.36)mm비(5.72±0.62)mm화(5.49 ±0.37)mm,P균＜0.05],대우가수술조(3.26 ±0.29) mm(P ＜0.05),LVESD현저소우보통선병독조화DMEM조[(3.87±0.45) mm비(4.91±0.62) mm화(4.63±0.37) mm,P균＜0.05],대우가수술조(1.93 ±0.25) mm(P ＜0.05).(3)혈관밀도검측결과:PDE5 shRNA조소서혈관밀도현저고우보통선병독조,심기경사구위(14.3±2.0)개비(6.6±1.2)개(P＜0.05)、경사주변위(23.6±2.1)개비(13.7±2.4)개(P＜0.05).(4) PDE5단백표체수평검측결과:PDE5 shRNA조현저저우보통선병독조,0.448 ±0.105비0.993±0.057(P ＜0.05).(5) cGMP화PKG단백표체적검측결과:PDE5 shRNA조량충단백적표체수평균현저고우대조조(P균＜0.05).결론 PDE5 shRNA가능통과상조cGMP화PKG표체감소소서심기경사면적화억제심기섬유화,종이억제심실중구、개선심공능.
    Objective To observe the impact of PDE5shRNA on cardiac remodeling and heart function following myocardial infarction in mice.Methods Myocardial infarction (MI) was induced in mice by left coronary artery ligation.Mice were randomly assigned to sham group (n =6),PDE5shRNA group (n =12),common adenovirus group (n =15) and DMEM group (n =8).Four weeks post-MI,the survival rate was evaluated.Cardiac function was examined by echocardiography.HE staining and Masson staining were used to evaluate the myocardial infarction size and fibrosis.The number of blood vessels was evaluated by immunohistochemistry,PDE5 protein expression in the left ventricular was detected using Western blot,level of cGMP or PKG activity in the left ventricle was evaluated with ELISA.Results Four weeks post-MI,all mice survived in the sham group,3(37％) mice died in the DMEM group,1 (8％) died in the PDE5shRNA group and 5 died in the common adenovirus group (33％).Infarct size was significantly reduced in PDE5shRNA group compared with the common adenovirus group and DMEM group [(25.4 ± 2.9) ％ vs.(42.0 ± 3.2) ％ and (43.4 ± 2.6) ％,P ＜ 0.05].Cardiac function was significantly improved in PDE5shRNA group compared to common adenovirus group and DMEM group[LVFS:(21.1 ± 3.7)％ vs.(14.2±2.9)％ and (14.22±2.91)％,allP＜0.05; LVEF:(48.2±7.1)％ vs.(34.6± 6.2)％ and (38.1 ±2.8)％,all P＜0.05; LVESD:(3.87 ±0.45) mm vs.(4.91 ±0.62) mm and (4.63 ± 0.37) mm,all P ＜ 0.05].The blood vessel density was also higher in PDE5shRNA group compared with common adenovirus group (infarct area:14.3 ± 2.0 vs.6.6 ± 1.2,P ＜ 0.05 ; periinfarct area:23.6 ±2.1 vs.13.7 ±2.4,P ＜0.05).Compared with common adenovirus group,level of PDE5 was significantly downregulated and level of cGMP or PKG was significantly upregulated in PDE5shRNA group (all P ＜ 0.05).Conclusions Present study suggests PDE5shRNA improves cardiac function and attenuates cardiac remodeling through reducing infarction size and cardiac fibrosis and these beneficial effects are possibly mediated by activating cGMP/PKG signaling pathway.
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