中华心律失常学杂志
中華心律失常學雜誌
중화심률실상학잡지
CHINESE JOURNAL OF CARDIAC ARRHYTHMIAS
2013年
4期
303-308
,共6页
张玲%孙娟%马嵋%侯月梅
張玲%孫娟%馬嵋%侯月梅
장령%손연%마미%후월매
微电极阵列%心肌梗死%场电位%组织工程心肌
微電極陣列%心肌梗死%場電位%組織工程心肌
미전겁진렬%심기경사%장전위%조직공정심기
Microelectrode arrays%Myocardial infarction%Field potential%Engineering heart tissue
目的 应用微电极阵列芯片(MEA)标测技术评价液态Ⅰ型胶原与新生大鼠心肌细胞混合构建工程化心肌组织(ECT)移植于大鼠陈旧性心肌梗死(MI)区后对心肌电重构的改善作用.方法 采用液态Ⅰ型胶原与新生大鼠心肌细胞混合培养构建ECT片层.48只成年雄性Wistar大鼠,随机分为4组:对照组、MI组、MI+片层移植组(MI+graft组)、MI+假片层移植组(MI+sham-gaft组),每组12只.对照组开胸不结扎动脉,其余各组结扎左冠状动脉前降支,建立大鼠MI模型.MI模型建立2周后行ECT移植.分别于术前及移植后2周行心电图及超声检测,采用主动脉逆行插管Langendorff法灌流心脏,MEA对ECT与宿主心肌之间的电偶联进行检测并记录心肌场电位.取心将ECT横切后采用免疫荧光染色心肌肌钙蛋白T(cTnT)、Ⅷ因子、连接蛋白43(Cx43),在激光扫描共焦显微镜下观察结果,采集图像数据.结果 移植的ECT紧密贴于MI部位,存活良好,有较丰富血管化的发生,在ECT中细胞间和与宿主之间形成了较为广泛的Cx43连接.MI+graft组较MI组左心室舒张末期容积缩小,左心室射血分数增加,但仍不能恢复正常.MI组术后4周较术前QRS时限、QT间期延长(P=0.01),MI+graft组比MI组QRS时限、QT间期缩短(P=0.03),仍比对照组长.MEA记录MI组MI区场电位振幅较对照组明显降低(P=0.01),时限明显延长(P=0.01);MI+graft组场电位较MI组振幅增加(P=0.01),时限缩短(P=0.02),但较对照组仍未完全恢复.对照组场电位激动传导呈顺序梯度传导.MI组传导不均一性增加,传导梯度样改变消失,MI+graft组使MI面心室肌电活动略呈均一性传导,但较对照组仍未完全恢复.结论 移植的ECT能改善MI大鼠心室肌的收缩功能及电传导时间,可部分修复心脏功能,组织工程化心肌组织有望用于心肌组织的缺损修复.
目的 應用微電極陣列芯片(MEA)標測技術評價液態Ⅰ型膠原與新生大鼠心肌細胞混閤構建工程化心肌組織(ECT)移植于大鼠陳舊性心肌梗死(MI)區後對心肌電重構的改善作用.方法 採用液態Ⅰ型膠原與新生大鼠心肌細胞混閤培養構建ECT片層.48隻成年雄性Wistar大鼠,隨機分為4組:對照組、MI組、MI+片層移植組(MI+graft組)、MI+假片層移植組(MI+sham-gaft組),每組12隻.對照組開胸不結扎動脈,其餘各組結扎左冠狀動脈前降支,建立大鼠MI模型.MI模型建立2週後行ECT移植.分彆于術前及移植後2週行心電圖及超聲檢測,採用主動脈逆行插管Langendorff法灌流心髒,MEA對ECT與宿主心肌之間的電偶聯進行檢測併記錄心肌場電位.取心將ECT橫切後採用免疫熒光染色心肌肌鈣蛋白T(cTnT)、Ⅷ因子、連接蛋白43(Cx43),在激光掃描共焦顯微鏡下觀察結果,採集圖像數據.結果 移植的ECT緊密貼于MI部位,存活良好,有較豐富血管化的髮生,在ECT中細胞間和與宿主之間形成瞭較為廣汎的Cx43連接.MI+graft組較MI組左心室舒張末期容積縮小,左心室射血分數增加,但仍不能恢複正常.MI組術後4週較術前QRS時限、QT間期延長(P=0.01),MI+graft組比MI組QRS時限、QT間期縮短(P=0.03),仍比對照組長.MEA記錄MI組MI區場電位振幅較對照組明顯降低(P=0.01),時限明顯延長(P=0.01);MI+graft組場電位較MI組振幅增加(P=0.01),時限縮短(P=0.02),但較對照組仍未完全恢複.對照組場電位激動傳導呈順序梯度傳導.MI組傳導不均一性增加,傳導梯度樣改變消失,MI+graft組使MI麵心室肌電活動略呈均一性傳導,但較對照組仍未完全恢複.結論 移植的ECT能改善MI大鼠心室肌的收縮功能及電傳導時間,可部分脩複心髒功能,組織工程化心肌組織有望用于心肌組織的缺損脩複.
목적 응용미전겁진렬심편(MEA)표측기술평개액태Ⅰ형효원여신생대서심기세포혼합구건공정화심기조직(ECT)이식우대서진구성심기경사(MI)구후대심기전중구적개선작용.방법 채용액태Ⅰ형효원여신생대서심기세포혼합배양구건ECT편층.48지성년웅성Wistar대서,수궤분위4조:대조조、MI조、MI+편층이식조(MI+graft조)、MI+가편층이식조(MI+sham-gaft조),매조12지.대조조개흉불결찰동맥,기여각조결찰좌관상동맥전강지,건립대서MI모형.MI모형건립2주후행ECT이식.분별우술전급이식후2주행심전도급초성검측,채용주동맥역행삽관Langendorff법관류심장,MEA대ECT여숙주심기지간적전우련진행검측병기록심기장전위.취심장ECT횡절후채용면역형광염색심기기개단백T(cTnT)、Ⅷ인자、련접단백43(Cx43),재격광소묘공초현미경하관찰결과,채집도상수거.결과 이식적ECT긴밀첩우MI부위,존활량호,유교봉부혈관화적발생,재ECT중세포간화여숙주지간형성료교위엄범적Cx43련접.MI+graft조교MI조좌심실서장말기용적축소,좌심실사혈분수증가,단잉불능회복정상.MI조술후4주교술전QRS시한、QT간기연장(P=0.01),MI+graft조비MI조QRS시한、QT간기축단(P=0.03),잉비대조조장.MEA기록MI조MI구장전위진폭교대조조명현강저(P=0.01),시한명현연장(P=0.01);MI+graft조장전위교MI조진폭증가(P=0.01),시한축단(P=0.02),단교대조조잉미완전회복.대조조장전위격동전도정순서제도전도.MI조전도불균일성증가,전도제도양개변소실,MI+graft조사MI면심실기전활동략정균일성전도,단교대조조잉미완전회복.결론 이식적ECT능개선MI대서심실기적수축공능급전전도시간,가부분수복심장공능,조직공정화심기조직유망용우심기조직적결손수복.
Objective To investigate improvement of the engineered cardiac tissue (ECT)constructed by neonatal rats and liquid collagen type I implantation on the myocardial electrophysiological characteristics after myocardial infarction(MI)in the rats by microelectrode array chip.Methods Engineered cardiac tissue was constructed by mixing cardiomyocytes isolated from the neonatal rats and liquid collagen type I as scaffold.Left anterior descending(LAD)coronary arteries of rats were ligatured to establish MI models.In control group thoracotomy and cardiac exposure was conducted but without LAD ligation.Forty-eight adult male Wistar rats were selected and divided randomly into four groups:control group (n =12),MI group(n =12) and MI+transplantation group(n=12),MI+sham-transplantation group(n =12).After four weeks,the ECG were recorded,and all hearts were intubated through aorta and perfused by the Langendorff apparatus with Tyrodes solution,and we recorded the field potential and electrical coupling between ECT and host with microeletrode arrays technique.cTnT,Ⅷ factor,connexin43 were measured by immunofluorescence staining,the results and collect images were observed under lasers scanning confocal microscopy.Results Transplanted ECT was covering the infarcted myocardium tightly,and had better biological activity.There were plenty of vascularization,and connexin43 was widely formed between ECT and the host.Echocardiography evaluation 4 weeks after infarction showed enlargement of left ventricular end-diastolic volume,decreased left ventricular ejection fraction.ECT reduced the enlargement of the volume LVED and increased the EF which were not restored to normal.Compared with the control group,ECG showed the expected QRS and QT interval prolonged in MI group(P=0.01).In MI+graft group QRS and QT interval shortened(P=0.03)but didn't restored to normal.We assessed electrical coupling of grafts to the host myocardium 4 weeks after engraftment by MEA.MI group showed the obvious delay of impulse propagation and decrease of amplitude in infarcted anterior and lateral segment (P=0.01).In contrast,epicardial activation in graft rats showed an increase in amplitude (P=0.01)and a shorten in field potential duration(P=0.02).Electricity impulse represented anisotropic alteration in MI group,and there was improvement in MI+graft group.Conclusion Engineered heart tissue grafts can improve electrical conduction and cardiac function as well as mechanical remodeling in infarcted rat hearts.Cardiac tissue engineering is a promising material for the repair of heart function and electrical injury in vivo.