目的 探讨MRI在研究停跳与不停跳两种心脏状态下,经猪心肌梗死模型心肌注射骨髓间充质干细胞后全身各主要脏器(心、肝、脾、肾)干细胞的早期再分布及疗效评价中的应用价值.方法 干细胞提取自雄性猪并用超顺磁性氧化铁标记.雌性猪制成急性心肌梗死模型7 d后数字表随机方法分为4组.第1组为心脏停跳细胞注射组(6只),体外循环后心脏停跳,超顺磁性氧化铁标记的干细胞(1×108)经心肌注入心肌梗死周边区.第2组为不停跳心脏细胞注射组(6只),相同的细胞在心脏跳动下经心肌注入心肌梗死周边区.第3组停跳心脏对照组(6只)和第4组不停跳心脏对照组(6只)中,相同剂量的生理盐水分别在停跳和不停跳状态下经心肌注入心肌梗死周边区.3 d后,行MR T2*WI示踪和心功能检查.并取动物心、肝、脾、肺、肾的标本,用实时定量聚合酶链反应(qRT-PCR)检测雄性细胞特异性的性别决定区域(SRY)基因定量检查.统计学分析采用方差分析和t检验.结果 第1～4组术前左心室舒张末期容积(LVEDV)分别为(56.8±5.3)、(54.8±6.8)、(57.4±4.3)和(56.8±2.8) ml,术后LVEDV分别为(65.2±5.2)、(63.2±3.7)、(60.2±4.7)和(62.2±4.4) ml,术前左心室收缩末期容积(LVESV)分别为(33.5±7.6)、(32.3±5.3)、(33.5±3.6)和(32.7±4.6) ml,术后LVESV分别为(37.3±5.6)、(36.3±6.9)、(34.3±5.4)和(36.3±8.1) ml;术前左心室射血分数值(LVEF)分别为(42.3±7.2)%、(41.7±6.8)%、(41.8±8.6)%和(42.7±7.7)%,术后LVEF分别为(44.5±8.7)%、(43.1±7.4)%、(42.8±5.6)%和(43.3±8.4)%;术前心肌梗死面积(MI)分别为(6.5±2.1)、(6.4±1.9)、(6.5±2.5)和(6.4±2.6) cm2,术后MI分别为(6.4±2.3)、(6.2±2.6)、(6.3±2.5)和(6.4±2.8) cm2,差异均无统计学意义(术前P值均＞0.05,F值分别为0.277、0.066、0.066、0.003;术后P值均＞0.05,F值分别为1.137、0.182、0.021及0.008).心脏中第1组较第2组T2*降低显著[(-22.3±2.2)和(-17.0±0.8) ms,t=-5.489,P＜0.01],而脾脏中第2组较第1组T2*值降低显著[(-7.7±0.7)和(-13.3±1.1) ms,t=9.055,P＜0.01],在肝脏及肾脏2组差异无统计学意义(肝脏t=-0.532,P>0.05,肾脏t=-0.113,P>0.05).SRY基因和qRT-PCR结果,第1组及第2组心脏[(150±62)和(72±4) U/L]、脾脏[(131±1)和(233±17) U/L]及肝脏[(17±1)和(9±5) U/L]差异有统计学意义(P值均＜0.05,t值分别为3.109、-13.286及3.492),病理学检查可以见移植细胞呈普鲁士蓝染色阳性,与MRI有很好的一致性.结论 干细胞氧化铁颗粒标记后,MRI可以作为方便而有效的手段在移植早期活体示踪干细胞,评价其在体内的分布情况.在心脏停跳状态下经心肌注射骨髓间充质干细胞将更有利于细胞在心脏的滞留.
目的 探討MRI在研究停跳與不停跳兩種心髒狀態下,經豬心肌梗死模型心肌註射骨髓間充質榦細胞後全身各主要髒器(心、肝、脾、腎)榦細胞的早期再分佈及療效評價中的應用價值.方法 榦細胞提取自雄性豬併用超順磁性氧化鐵標記.雌性豬製成急性心肌梗死模型7 d後數字錶隨機方法分為4組.第1組為心髒停跳細胞註射組(6隻),體外循環後心髒停跳,超順磁性氧化鐵標記的榦細胞(1×108)經心肌註入心肌梗死週邊區.第2組為不停跳心髒細胞註射組(6隻),相同的細胞在心髒跳動下經心肌註入心肌梗死週邊區.第3組停跳心髒對照組(6隻)和第4組不停跳心髒對照組(6隻)中,相同劑量的生理鹽水分彆在停跳和不停跳狀態下經心肌註入心肌梗死週邊區.3 d後,行MR T2*WI示蹤和心功能檢查.併取動物心、肝、脾、肺、腎的標本,用實時定量聚閤酶鏈反應(qRT-PCR)檢測雄性細胞特異性的性彆決定區域(SRY)基因定量檢查.統計學分析採用方差分析和t檢驗.結果 第1～4組術前左心室舒張末期容積(LVEDV)分彆為(56.8±5.3)、(54.8±6.8)、(57.4±4.3)和(56.8±2.8) ml,術後LVEDV分彆為(65.2±5.2)、(63.2±3.7)、(60.2±4.7)和(62.2±4.4) ml,術前左心室收縮末期容積(LVESV)分彆為(33.5±7.6)、(32.3±5.3)、(33.5±3.6)和(32.7±4.6) ml,術後LVESV分彆為(37.3±5.6)、(36.3±6.9)、(34.3±5.4)和(36.3±8.1) ml;術前左心室射血分數值(LVEF)分彆為(42.3±7.2)%、(41.7±6.8)%、(41.8±8.6)%和(42.7±7.7)%,術後LVEF分彆為(44.5±8.7)%、(43.1±7.4)%、(42.8±5.6)%和(43.3±8.4)%;術前心肌梗死麵積(MI)分彆為(6.5±2.1)、(6.4±1.9)、(6.5±2.5)和(6.4±2.6) cm2,術後MI分彆為(6.4±2.3)、(6.2±2.6)、(6.3±2.5)和(6.4±2.8) cm2,差異均無統計學意義(術前P值均＞0.05,F值分彆為0.277、0.066、0.066、0.003;術後P值均＞0.05,F值分彆為1.137、0.182、0.021及0.008).心髒中第1組較第2組T2*降低顯著[(-22.3±2.2)和(-17.0±0.8) ms,t=-5.489,P＜0.01],而脾髒中第2組較第1組T2*值降低顯著[(-7.7±0.7)和(-13.3±1.1) ms,t=9.055,P＜0.01],在肝髒及腎髒2組差異無統計學意義(肝髒t=-0.532,P>0.05,腎髒t=-0.113,P>0.05).SRY基因和qRT-PCR結果,第1組及第2組心髒[(150±62)和(72±4) U/L]、脾髒[(131±1)和(233±17) U/L]及肝髒[(17±1)和(9±5) U/L]差異有統計學意義(P值均＜0.05,t值分彆為3.109、-13.286及3.492),病理學檢查可以見移植細胞呈普魯士藍染色暘性,與MRI有很好的一緻性.結論 榦細胞氧化鐵顆粒標記後,MRI可以作為方便而有效的手段在移植早期活體示蹤榦細胞,評價其在體內的分佈情況.在心髒停跳狀態下經心肌註射骨髓間充質榦細胞將更有利于細胞在心髒的滯留.
목적 탐토MRI재연구정도여불정도량충심장상태하,경저심기경사모형심기주사골수간충질간세포후전신각주요장기(심、간、비、신)간세포적조기재분포급료효평개중적응용개치.방법 간세포제취자웅성저병용초순자성양화철표기.자성저제성급성심기경사모형7 d후수자표수궤방법분위4조.제1조위심장정도세포주사조(6지),체외순배후심장정도,초순자성양화철표기적간세포(1×108)경심기주입심기경사주변구.제2조위불정도심장세포주사조(6지),상동적세포재심장도동하경심기주입심기경사주변구.제3조정도심장대조조(6지)화제4조불정도심장대조조(6지)중,상동제량적생리염수분별재정도화불정도상태하경심기주입심기경사주변구.3 d후,행MR T2*WI시종화심공능검사.병취동물심、간、비、폐、신적표본,용실시정량취합매련반응(qRT-PCR)검측웅성세포특이성적성별결정구역(SRY)기인정량검사.통계학분석채용방차분석화t검험.결과 제1～4조술전좌심실서장말기용적(LVEDV)분별위(56.8±5.3)、(54.8±6.8)、(57.4±4.3)화(56.8±2.8) ml,술후LVEDV분별위(65.2±5.2)、(63.2±3.7)、(60.2±4.7)화(62.2±4.4) ml,술전좌심실수축말기용적(LVESV)분별위(33.5±7.6)、(32.3±5.3)、(33.5±3.6)화(32.7±4.6) ml,술후LVESV분별위(37.3±5.6)、(36.3±6.9)、(34.3±5.4)화(36.3±8.1) ml;술전좌심실사혈분수치(LVEF)분별위(42.3±7.2)%、(41.7±6.8)%、(41.8±8.6)%화(42.7±7.7)%,술후LVEF분별위(44.5±8.7)%、(43.1±7.4)%、(42.8±5.6)%화(43.3±8.4)%;술전심기경사면적(MI)분별위(6.5±2.1)、(6.4±1.9)、(6.5±2.5)화(6.4±2.6) cm2,술후MI분별위(6.4±2.3)、(6.2±2.6)、(6.3±2.5)화(6.4±2.8) cm2,차이균무통계학의의(술전P치균＞0.05,F치분별위0.277、0.066、0.066、0.003;술후P치균＞0.05,F치분별위1.137、0.182、0.021급0.008).심장중제1조교제2조T2*강저현저[(-22.3±2.2)화(-17.0±0.8) ms,t=-5.489,P＜0.01],이비장중제2조교제1조T2*치강저현저[(-7.7±0.7)화(-13.3±1.1) ms,t=9.055,P＜0.01],재간장급신장2조차이무통계학의의(간장t=-0.532,P>0.05,신장t=-0.113,P>0.05).SRY기인화qRT-PCR결과,제1조급제2조심장[(150±62)화(72±4) U/L]、비장[(131±1)화(233±17) U/L]급간장[(17±1)화(9±5) U/L]차이유통계학의의(P치균＜0.05,t치분별위3.109、-13.286급3.492),병이학검사가이견이식세포정보로사람염색양성,여MRI유흔호적일치성.결론 간세포양화철과립표기후,MRI가이작위방편이유효적수단재이식조기활체시종간세포,평개기재체내적분포정황.재심장정도상태하경심기주사골수간충질간세포장경유리우세포재심장적체류.
Objective To evaluate the efficacy of MRI for assessment of re-distribution of bone marrow mesenchymal stem cells injected intramyocardially in main organs (heart, liver, spleen and kidney) under different heart status (beating or arresting) in a porcine model. Methods Bone marrow-derived mesenchymal stem cells were obtained from the male swine and labeled with iron oxide during culture. Acute myocardial infarction was created in female swine, one week later, the survivors were randomly divided into 4 groups. Cardiopulmonary bypass was set up to arrest the heart, and then labeled cells (1×108) were intramyocardially injected into the border of the infracted myocardium in group 1 (n=6). The same volume of cells was grafted into the beating heart in group 2 (n=6). In group 3 and 4, saline was injected into either the arresting or beating myocardium. Three days later, re-distribution of stem cells and cardiac function were assessed by T2*WI and cine MRI, respectively. All animals were sacrificed for histology and real-time quantitative polymerase chain reaction (RT-PCR) of sex-determining region on Y-chromosome (SRY) investigation.The ANOVA and t test was used for statistics. Results The left ventricular end-diastolic volume (LVEDV) before transplantation for group 1-4 were: (56.8±5.3),(54.8±6.8),(57.4±4.3)and(56.8±2.8) ml, and after transplantation for group 1-4 were: (65.2±5.2),(63.2±3.7),(60.2±4.7)and(62.2±4.4) ml. The left ventricular end-systolic volume (LVESV) before transplantation for group 1-4 were: (33.5±7.6),(32.3±5.3),(33.5±3.6)and(32.7±4.6) ml,and after transplantation for group 1-4 were: (37.3±5.6),(36.3±6.9),(34.3±5.4)and(36.3±8.1) ml. The left ventricular EF values (LVEF) before transplantation for group 1-4 were: (42.3±7.2)%,(41.7±6.8)%,(41.8±8.6)% and(42.7±7.7)%,and after transplantation for group 1-4 were: (44.5±8.7)%,(43.1±7.4)%,(42.8±5.6)% and(43.3±8.4)%. The myocardial infarction area (MI) before transplantation for group 1-4 were: (6.5±2.1),(6.4±1.9),(6.5±2.5)and(6.4±2.6) cm2,and after transplantation for group 1-4 were: (6.4±2.3),(6.2±2.6),(6.3±2.5)and(6.4±2.8) cm2 . There were no statistical differences before and after transplantation in these 4 groups[P values of before and after transplantation for LVEDV, LVESV, LVEF,MI were >0.05 (F= 0.277, 0.066,0.066, 0.003); and >0.05 (F= 1.137,0.182,0.021,0.008),respectively]. The T2 value of the infracted myocardium in group 1 decreased more obviously than that in group 2[(-22.3 ± 2.2) vs (-17.0 ± 0.8) ms, t=-5.489, P＜0.01], while the T2 value of the spleen decreased more significantly in group 2 than that in group 1[(-7.7 ± 0.7) vs (-13.3 ± 1.1) ms,t=9.055, P＜0.01]. The T2 values of the liver and kidney were no significant differences in group 1 and 2 (liver, t=-0.532,P>0.05 and kidney, t=-0.113,P>0.05). The results of RT-PCR in group 1 and 2 showed significant differences in heart[(150±62) vs (72±4) U/L ,P<0.05, t=3.109], spleen[(131±1) vs (233±17) U/L, P＜0.01, t=- 13.286]and liver[(17±1) vs (9±5) U/L ,P<0.01,t= 3.492]. Pathological examination demonstrated that the transplanted stem cells were positive for Prussian blue staining, which had a good correlation with MRI results. Conclusion MRI can serve as a convenient and efficient imaging method to track the migration of stem cells with SPIO labeled in early stage and evaluate its early re-distribution in vivo. Injection of bone marrow mesenchymal stem cells in the arresting heart could favor retaining more cells in the myocardium.
