CAJ | 학술논문

目的探讨纤维蛋白胶联合大鼠脂肪干细胞(ADSCs)移植于梗死心肌后的细胞存活及对心功能的影响.方法分离培养大鼠脂肪干细胞(ADSCs),流式细胞术鉴定其表型.将雌性SD大鼠左冠状动脉前降支结扎建立心肌梗死模型,模型建立1周后,存活大鼠随机(随机数字法)分成4组,即对照组(n=10)、纤维蛋白胶组(n=10)、细胞组(n=10)及联合组(n=10).于梗死边缘区分别移植100μl PBS、纤维蛋白胶和4,6二乙酰基-2-苯基吲哚(4,6-diamidino-2-phenylindole DAPI)标记的100μl ADSCs,联合组用纤维蛋白胶加ADSCs注射.移植后4周,用血流动力学评估心功能,后处死动物取心肌标本,测量心肌梗死面积；用DAPI、肌节辅肌动蛋白免疫组化双染法确定细胞的存活；Ⅷ因子染色法检测血管新生.数据用SPSS 15.0统计软件统汁分析(多组间比较采用单因素方差分析,两组间比较采用成组t检验).以P＜ 0.05为差异具有统计学意义.结果细胞移植后4周,联合组的LVSP,+dp/dtmax均高于其他三组(P＜0.05).联合组心肌梗死面积(28.5±3.6)％明显低于细胞组(33.33±2.3)％和纤维蛋白胶组(35.96±2.11)％,(均P ＜0.05).联合组梗死区的毛细血管密度(108.7±11.38/mm2)显著高于细胞组、纤维蛋白胶组和对照组(均P＜0.01).用DAPI、肌节辅肌动蛋白免疫组化双染法示联合组心肌梗死区的存活细胞数量多于单纯细胞移植组.结论纤维蛋白胶联合ADSCs移植可以提高细胞的存活并显著改善大鼠心肌梗死后的心功能.
목적 탐토섬유단백효연합대서지방간세포(ADSCs)이식우경사심기후적세포존활급대심공능적영향.방법 분리배양대서지방간세포(ADSCs),류식세포술감정기표형.장자성SD대서좌관상동맥전강지결찰건립심기경사모형,모형건립1주후,존활대서수궤(수궤수자법)분성4조,즉대조조(n=10)、섬유단백효조(n=10)、세포조(n=10)급연합조(n=10).우경사변연구분별이식100μl PBS、섬유단백효화4,6이을선기-2-분기신타(4,6-diamidino-2-phenylindole DAPI)표기적100μl ADSCs,연합조용섬유단백효가ADSCs주사.이식후4주,용혈류동역학평고심공능,후처사동물취심기표본,측량심기경사면적；용DAPI、기절보기동단백면역조화쌍염법학정세포적존활；Ⅷ인자염색법검측혈관신생.수거용SPSS 15.0통계연건통즙분석(다조간비교채용단인소방차분석,량조간비교채용성조t검험).이P＜ 0.05위차이구유통계학의의.결과 세포이식후4주,연합조적LVSP,+dp/dtmax균고우기타삼조(P＜0.05).연합조심기경사면적(28.5±3.6)％명현저우세포조(33.33±2.3)％화섬유단백효조(35.96±2.11)％,(균P ＜0.05).연합조경사구적모세혈관밀도(108.7±11.38/mm2)현저고우세포조、섬유단백효조화대조조(균P＜0.01).용DAPI、기절보기동단백면역조화쌍염법시연합조심기경사구적존활세포수량다우단순세포이식조.결론 섬유단백효연합ADSCs이식가이제고세포적존활병현저개선대서심기경사후적심공능.
Objective To investigate the cell survival of the combination of fibrin glue and adiposederived stem cells (ADSCs) in rats when implanted into ischemic myocardium and the improvement of heart function.Methods The rat ADSCs were isolated from the subcutaneous adipose tissues.The surface phenotype of these cells was analyzed by flow cytometry.Myocardial infarction was induced in female rats using coronary artery ligation.One week after MI,surviving rats were randomized (random nuber) into 4 groups,control group (n =10),fibrin group (n =10),cell group (n =10) and combination group (n =10).100 μl of PBS was injected into the ischemic myocardium in control group.100 μl of Fibrin glue were injected into ischemic myocardium in fibrin group.100 μl of ADSCs labeled with DAPI were injected into the infract along the border zone in cell group.ADSCs in 100 μl of fibrin glue were injected into the infract in combination group.Four weeks after the injection the surviving rats underwent examination of heart functions by the Hemodynamics.The rats were killed and their hearts were taken out to undergo immunohistochemistry with 4,6-diamidino-2-phenylindole (DAPI) and actin and factor Ⅶ to measure the area of cardiac infarction and the capillary density.The heart infarcted size was calculated by masson trichrome staining.All data was analyzed by software SPSS 15.0,ANOVA comparison tests and the student t test were used,and P ＜ 0.05 was considered as statistically significant.Results Four weeks after the cells were transplanted,LVSP and + dp/dtmax of combination group were highest among all groups.The heart infarcted size of the combination group was (28.5 ± 3.6) ％,significantly less than those of the cell group (33.33 ± 2.3) ％ and fibrin group (35.96 ± 2.11) ％,both P ＜ 0.05.The capillary density of the combination group was (108.7 ± 11.38) /mm2,significantly greater than those of the cell group and that of the fibrin group,and greater than that of the control group.DAPI and actin double staining detected a varied increase in the number of surviving cardiomyoctyes at the heart infarcted area.Conclusions Transplantation of ADSCs with fibrin glue brings better improvement in cell survival and in restoration of heart function than either cellular or fibrin therapy alone.