摘要
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SPSS15．0统计软件统计分析（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／mm^2）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。
Objective To investigate the cell survival of the combination of fibrin glue and adipose- derived stem cells （ADSCs） in rats when implanted into ischemic myocardinm 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 myocardinm 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 heartfunctions 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 VIH 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/dt 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） /mm^2, 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.
出处
《中华急诊医学杂志》
CAS
CSCD
北大核心
2012年第12期1322-1326,共5页
Chinese Journal of Emergency Medicine
基金
新疆维吾尔自治区科技支疆项目（200891127）
关键词
脂肪干细胞
纤维蛋白胶
细胞移植
急性心肌梗死
大鼠
Adipose tissue-derived stem cells
Fibrin
Cell transplantation
Acute myocardial infarction
Rat
