不同途径移植人羊膜上皮细胞对心肌梗死大鼠心脏的保护作用  

Protective effects of different transplantations of human amniotic epithelial cells in rat heart of myocardial infarction

作　　者：王钰莹[1] 方宁[1] 陈代雄[1] 余丽梅[1] 章涛[1] 赵春华[2] 

机构地区：[1]遵义医学院附属医院贵州省细胞工程重点实验室,贵州遵义563003 [2]中国医学科学院基础医学研究所组织工程研究中心,北京100005

出　　处：《中国新药与临床杂志》2013年第6期476-481,共6页Chinese Journal of New Drugs and Clinical Remedies

基　　金："重大新药创制"国家科技重大专项课题(2009ZX09503-025);贵州省科技厅公关项目(黔科合SZ字[2009]3017)

摘　　要：目的比较经舌下静脉和心外膜直视注射移植人羊膜上皮细胞(hAECs)对心肌梗死大鼠的心脏保护作用。方法雄性SD大鼠48只,随机分为假手术组(n=12)、模型组(n=12)、hAECs心外膜移植组(n=12)和hAECs静脉移植组(n=12),结扎大鼠左冠状动脉前降支制备心肌梗死模型。hAECs经体外分离、纯化、培养和标记后,在心肌梗死后1周,分别经心外膜和舌下静脉注射hAECs,4周后测定心功能,免疫组化法检测Ⅷ因子相关抗原(vWF)的表达,免疫荧光观察移植细胞的定植和存活。结果免疫荧光染色可见两种途径移植组心肌组织中均有DAPI标记细胞存在;免疫组化结果显示,200倍视野下hAECs静脉移植组和hAECs心外膜移植组的vWF染色阳性微血管计数较模型组明显增高[(22.00±2.61)和(19.80±2.32)vs.(9.80±1.17),P<0.01]。与模型组相比,两移植组左心功能显著改善(P<0.05)。结论 hAECs经静脉移植和心外膜移植均可归巢至心肌梗死区,并促进新生血管形成,对梗死大鼠的心脏具有保护作用。AIM To study the protective effects of intravenous and via epicardium transplantation of human amniotic epithelial cells （hAECs） against rat myocardial infarction （MI）. METHODS Forty-eight male Sprague-Dawley rats were randomly assigned into sham group （n = 12）, model group （n = 14）, hAECs epicardial transplantation group （n = 14） and hAECs vein graft group （n -- 13）. MI was established by occluding the left anterior descending artery （LAD） in SD rats. The hAECs isolated from human amnion by trypsin digestion, were amplificated and labeled with 4, 6-diamidino-2 phenylindole （DAPI）. One week after MI, the labeled hAECs were injected into sublingual vein or epicardium of model rats. The heart functions were evaluated after four weeks of the treatment, immunohistochemistry was used to determine the expression of Ⅷ- yon Willebrand factor （Ⅷ） and immunofluorescence was employed to observe the survival and integration of the transplanted cells. RESULTS DAPI positive cells were found in MI region on two transplantation groups by immunofluorescence staining. The labeled cells were visible in the host hearts, immunochemical staining revealed that the engrafted stem cells expressed vWF. The microvessel density （MVD） in the infarction regions of both the hAECs vein graft group and the hAECs epicardial injection group were significant higher than in the model group [（22.00 ± 2.61） and （19.80 ±2.32） vs. （9.80 ± 1.17）, P 〈 0.01]. Left heart functions and infarct size in two transplantation groups were obviously improved compared with those of the model group （P 〈 0.01）. CONCLUSION Both intravenous and via epieardium transplanted hAECs are able to home into the infarcted areas, and improves the heart function by promoting the revascularization in this regions.

关 键 词：人羊膜上皮细胞 心肌梗死 细胞移植 心脏保护 

分 类 号：R394.2[医药卫生—医学遗传学]