中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2012年
12期
1045-1050
,共6页
杜优优%姚瑞%蒲实%赵晓燕%刘光辉%赵洛沙%陈庆华%李凌
杜優優%姚瑞%蒲實%趙曉燕%劉光輝%趙洛沙%陳慶華%李凌
두우우%요서%포실%조효연%류광휘%조락사%진경화%리릉
心肌梗死%间质干细胞%心室重构%肌纤维母细胞
心肌梗死%間質榦細胞%心室重構%肌纖維母細胞
심기경사%간질간세포%심실중구%기섬유모세포
Myocardial infarction%Mesenchymal stem cells%Ventricular remodeling%Myofibroblasts
目的 探讨骨髓间充质干细胞(MSC)对梗死心肌中肌纤维母细胞聚集的调节作用.方法 采用结扎大鼠冠状动脉前降支的方法复制心肌梗死(MI)模型,实验动物应用随机数字表进行完全随机化分组,分为假手术组(仅穿线不结扎冠状动脉,n=8)、心肌梗死+磷酸盐缓冲液(MI+PBS)组(结扎冠状动脉+心肌注射PBS溶液,n=8)和MI+ MSC组(结扎冠状动脉+心肌注射MSC,n=8).通过心脏超声检查、血液动力学检查和Masson三色染色等方法分别检测左心室射血分数(LVEF)、短轴缩短率(FS)、左心室收缩末压力(LVSP)、左心室舒张末压力(LVEDP)、左心室压最大升降速率(±dp/dtmax)、MI面积、梗死区室壁厚度和心脏质量/体质量等指标,评价MSC对大鼠心脏功能及心室重构的影响.分别采用荧光免疫组化染色、实时定量逆转录PCR、Western blot等方法检测肌纤维母细胞向梗死区聚集、移植MSC存活和分化、转化生长因子β1(TGF-β1)-Smad2信号通路激活等情况.结果 (1)心室重构和心脏功能指标检测结果:术后28 d,与MI+ PBS组比较,MI+ MSC组的心脏质量/体质量[(3.04 ±0.16)mg/g比(3.34 ±0.14) mg/g,P＜0.01]及心脏梗死面积[(38.72±2.38)％比(46.36±2.81)％,P＜0.01]较低；梗死区室壁厚度较大[(0.93 ±0.17)mm比(0.65±0.16) mm,P=0.01];LVEF[(32.5±5.9)％比(26.5±4.5)％,P=0.03]及FS[(30.4±3.3)％比(24.8±4.2)％,P＜0.01]较高；改善血液动力学参数,LVSP较高(P=0.01),LVEDP较低(P＜0.01).(2)与MI+ PBS组比较,MI+ MSC组大鼠心脏梗死区肌纤维母细胞聚集数量较高[(196±20)/mm2比(89 ±25)/mm2,P＜0.01],其中部分绿色荧光蛋白(GFP)+移植细胞同时表达α-SMA+.(3)与注射PBS比较,移植MSC可以明显增加梗死区TGF-β1 mRNA和蛋白表达水平(P均＜0.01),促进Smad2的磷酸化(P＜0.01).结论 MSC可能通过直接分化和激活TGF-β1-Smad2信号通路促进成纤维母细胞向肌纤维母细胞转型增加大鼠心脏梗死区肌纤维母细胞的聚集,从而抑制心室重构,改善心脏功能.
目的 探討骨髓間充質榦細胞(MSC)對梗死心肌中肌纖維母細胞聚集的調節作用.方法 採用結扎大鼠冠狀動脈前降支的方法複製心肌梗死(MI)模型,實驗動物應用隨機數字錶進行完全隨機化分組,分為假手術組(僅穿線不結扎冠狀動脈,n=8)、心肌梗死+燐痠鹽緩遲液(MI+PBS)組(結扎冠狀動脈+心肌註射PBS溶液,n=8)和MI+ MSC組(結扎冠狀動脈+心肌註射MSC,n=8).通過心髒超聲檢查、血液動力學檢查和Masson三色染色等方法分彆檢測左心室射血分數(LVEF)、短軸縮短率(FS)、左心室收縮末壓力(LVSP)、左心室舒張末壓力(LVEDP)、左心室壓最大升降速率(±dp/dtmax)、MI麵積、梗死區室壁厚度和心髒質量/體質量等指標,評價MSC對大鼠心髒功能及心室重構的影響.分彆採用熒光免疫組化染色、實時定量逆轉錄PCR、Western blot等方法檢測肌纖維母細胞嚮梗死區聚集、移植MSC存活和分化、轉化生長因子β1(TGF-β1)-Smad2信號通路激活等情況.結果 (1)心室重構和心髒功能指標檢測結果:術後28 d,與MI+ PBS組比較,MI+ MSC組的心髒質量/體質量[(3.04 ±0.16)mg/g比(3.34 ±0.14) mg/g,P＜0.01]及心髒梗死麵積[(38.72±2.38)％比(46.36±2.81)％,P＜0.01]較低；梗死區室壁厚度較大[(0.93 ±0.17)mm比(0.65±0.16) mm,P=0.01];LVEF[(32.5±5.9)％比(26.5±4.5)％,P=0.03]及FS[(30.4±3.3)％比(24.8±4.2)％,P＜0.01]較高；改善血液動力學參數,LVSP較高(P=0.01),LVEDP較低(P＜0.01).(2)與MI+ PBS組比較,MI+ MSC組大鼠心髒梗死區肌纖維母細胞聚集數量較高[(196±20)/mm2比(89 ±25)/mm2,P＜0.01],其中部分綠色熒光蛋白(GFP)+移植細胞同時錶達α-SMA+.(3)與註射PBS比較,移植MSC可以明顯增加梗死區TGF-β1 mRNA和蛋白錶達水平(P均＜0.01),促進Smad2的燐痠化(P＜0.01).結論 MSC可能通過直接分化和激活TGF-β1-Smad2信號通路促進成纖維母細胞嚮肌纖維母細胞轉型增加大鼠心髒梗死區肌纖維母細胞的聚集,從而抑製心室重構,改善心髒功能.
목적 탐토골수간충질간세포(MSC)대경사심기중기섬유모세포취집적조절작용.방법 채용결찰대서관상동맥전강지적방법복제심기경사(MI)모형,실험동물응용수궤수자표진행완전수궤화분조,분위가수술조(부천선불결찰관상동맥,n=8)、심기경사+린산염완충액(MI+PBS)조(결찰관상동맥+심기주사PBS용액,n=8)화MI+ MSC조(결찰관상동맥+심기주사MSC,n=8).통과심장초성검사、혈액동역학검사화Masson삼색염색등방법분별검측좌심실사혈분수(LVEF)、단축축단솔(FS)、좌심실수축말압력(LVSP)、좌심실서장말압력(LVEDP)、좌심실압최대승강속솔(±dp/dtmax)、MI면적、경사구실벽후도화심장질량/체질량등지표,평개MSC대대서심장공능급심실중구적영향.분별채용형광면역조화염색、실시정량역전록PCR、Western blot등방법검측기섬유모세포향경사구취집、이식MSC존활화분화、전화생장인자β1(TGF-β1)-Smad2신호통로격활등정황.결과 (1)심실중구화심장공능지표검측결과:술후28 d,여MI+ PBS조비교,MI+ MSC조적심장질량/체질량[(3.04 ±0.16)mg/g비(3.34 ±0.14) mg/g,P＜0.01]급심장경사면적[(38.72±2.38)％비(46.36±2.81)％,P＜0.01]교저；경사구실벽후도교대[(0.93 ±0.17)mm비(0.65±0.16) mm,P=0.01];LVEF[(32.5±5.9)％비(26.5±4.5)％,P=0.03]급FS[(30.4±3.3)％비(24.8±4.2)％,P＜0.01]교고；개선혈액동역학삼수,LVSP교고(P=0.01),LVEDP교저(P＜0.01).(2)여MI+ PBS조비교,MI+ MSC조대서심장경사구기섬유모세포취집수량교고[(196±20)/mm2비(89 ±25)/mm2,P＜0.01],기중부분록색형광단백(GFP)+이식세포동시표체α-SMA+.(3)여주사PBS비교,이식MSC가이명현증가경사구TGF-β1 mRNA화단백표체수평(P균＜0.01),촉진Smad2적린산화(P＜0.01).결론 MSC가능통과직접분화화격활TGF-β1-Smad2신호통로촉진성섬유모세포향기섬유모세포전형증가대서심장경사구기섬유모세포적취집,종이억제심실중구,개선심장공능.
Objective To investigate the modulation effects of mesenchymal stem cells (MSC) implantation on the myofibroblasts congregating in the infarct region after myocardial infarction (MI).Methods MI was induced in SD rats by left anterior descending coronary artery ligation,and the experimental animals were assigned randomly into the sham group,MI + PBS group and MI + MSC group (myocardial injection of 0.1 ml 2 × 107/ml in four locations in the infarct region).Echocardiography,hemodynamic examinations and Masson trichrome staining were performed.Implanted MSC differentiation and myofibroblasts congregating in infarct region were investigated by immunofluorescence staining.TGF-β1-Smad2 signaling pathway was examined by real-time RT-PCR and Western blot.Results (1) Four weeks late,heart-weight/body-weight ratio [(3.04 ± 0.16) mg/g vs.(3.34 ± 0.14) mg/g,P ＜ 0.01] and myocardial infarction size [(38.72 ± 2.38) ％ vs.(46.36 ± 2.81) ％,P ＜ 0.01] were significantly reduced in MI + MSC group than in MI + PBS group,while scar thickness of infarct region was thicker [(0.93 ±0.17) mm vs.(0.65 ±0.16) mm,P=0.01],and LVEF was higher [LVEF:(32.5 ±5.9)％ vs.(26.5 ±4.5) ％,P =0.03] in MI + MSC group than in MI + PBS group.(2) Myofibroblasts congregating in the infarct region was significantly enhanced in MI + MSC group compared with MI + PBS group [(196 ± 20) cells/mm2 vs.(89 ± 25) cells/mm2,P ＜ 0.01],and part of implanted MSC expressed α-SMA +.(3) TGF-β1 expression and the phosphorylating of Smad2 in the infarct region were significantly upregulated in MI + MSC group compared with MI + PBS group (all P ＜ 0.05).Conclusions MSC could improve myocardial function and promote myofibroblasts congregating in the infarct region via activating the TGF-β1-Smad2 signaling pathway in this model.
