摘要
背景:研究发现促红细胞生成素尚具有多种非血液系统生理效应,如参与哺乳动物胚胎的正常发育、抑制炎症、促进血管生长,以及在多种组织和细胞损伤中抑制细胞凋亡、减少梗死面积、促进功能恢复等作用。目的:观察重组人促红细胞生成素与骨骼肌卫星细胞联合移植对兔急性心肌梗死是否具有积极作用?设计、时间及地点:随机对照动物实验,于2006-01/07在深圳市人民医院中心实验室完成。材料:自体移植:清洁级雄性新西兰大白兔50只,随机分为假手术组、模型对照组、细胞移植组、重组人促红细胞生成素组、联合组,10只/组。异体异性移植:同品系雄性新西兰大白兔10只,作为供体用于制备骨骼肌卫星细胞;另取雌性兔10只作为受体,分为细胞移植组、联合组,5只/组。重组人促红细胞生成素为沈阳三生制药股份有限公司产品。方法:自体移植实验:除假手术组外,其余4组建立急性心肌梗死模型。重组人促红细胞生成素组、联合组在冠脉结扎同时,采用肌注法一次性给予重组人促红细胞生成素3000U/kg;细胞移植组、联合组将含有约107个骨骼肌卫星细胞的DMEM溶液200μL多点直接注射在心脏表面梗死区。异体异性移植实验:细胞移植组、联合组均建立急性心肌梗死模型,根据分组要求注射骨骼肌卫星细胞和重组人促红细胞生成素,浓度及方法同上,5d后取材行PCR检测。主要观察指标:测定心肌梗死面积,免疫组织化学法检测心肌组织血运改善及desmin表达,荧光检测骨骼肌卫星细胞,SRY基因PCR检测结果。结果:移植后4周与模型对照组比较,细胞移植组、重组人促红细胞生成素组、联合组心肌梗死面积均明显减小(P<0.05),且联合组减小幅度强于另外两组(P<0.05)。各组梗死中心区CD34阳性细胞数量与上面结果相似。细胞移植组、联合组心肌组织desmin呈阳性表达,其余3组呈阴性。联合组移植区DAPI标记的蓝色荧光细胞数较细胞移植组明显增多。异体异性移植实验中,联合组SRY基因扩增条带的宽度和亮度均明显强于细胞移植组。结论:重组人促红细胞生成素与骨骼肌卫星细胞联合移植能够进一步缩小兔心肌梗死面积,改善梗死区血运,提高骨骼肌卫星细胞移植后的存活率,二者对治疗急性心肌梗死具有协同作用。
BACKGROUND: Erythropoietin has many physiological effect of non-hematological system, such as participating in normal development of mammal embryo, inhibiting inflammation, enhancing vessel growth, as well as inhibiting apoptosis, reducing infarcted area and accelerating functional recovery in injuries to many tissues and cells.
OBJECTIVE: To study whether recombinant human erythropoietin (rhEPO) can cooperate with skeletal muscle satellite cells to cure acute myocardial infarction rabbit.
DESIGN, TIME AND SETTING: The randomized control animal experiment was performed at the Central Laboratory of Shenzben People's Hospital from January to July 2006.
MATERIALS: Autologous transplantation: Fifty New Zealand male rabbits were randomly assigned into sham operation group, model control group, cell transplantation group, rhEPO group and combination group with 10 rabbits in each group. Allograft opposite sex transplantation: Ten New Zealand male rabbits of the same strain (donors) were used to prepare skeletal muscle satellite cells. An additional ten female rabbits (recipients) were allocated into five groups, rhEPO was purchased from Shenyang Sunshine Pharmaceutical Co., Ltd.
METHODS: Autologous transplantation: Rabbit models of acute myocardial infarction were established in the model control group, cell transplantation group, rhEPO group and combination group. 3 000 U/kg rhEPO was injected by muscle injection method when ligating coronary artery in the rhEPO group and combination group. 200 g L of DMEM supplemented with about 107 skeletal muscle satellite cells was infused into the infarcted areas in the cell transplantation group and combination group. Allograft opposite sex transplantation: Rabbit models of acute myocardial infarction were created in the cell transplantation group and combination group. Skeletal muscle satellite cells and rhEPO were infused according to the requirement of each group. The concentration and method were the same as above procedures. Polymerase chain reaction was used 5 days later.
MAIN OUTCOME MEASURES: Infarcted area was measured. Immunohistochemistry was used to identify blood transportation and desmin expression of myocardial tissues. Fluorescence was applied to examine skeletal muscle satellite cells. Polymerase chain reaction was used to proliferate the gene SRY.
RESULTS: At 4 weeks after transplantation, infarcted area significantly reduced in the cell transplantation group, rhEPO group and combination group, especially in the combination group, compared with the model control group (P 〈 0.05). The number of C34 positive cells was similar to above-mentioned results. The desmin was positively expressed in the myocardial tissues in the cell transplantation group and combination group, but negatively in the sham operation group, model control group and rhEPO group. DAPI-positive cells were more in the combination group than in the cell transplantation group. In allograft opposite sex transplantation, the width and brightness of SRY gene amplified band were significantly stronger in the combination group than in the cell transplantation group.
CONCLUSION: Combined transplantation of rhEPO and akeletal muscle satellite cells can shorten rabbit myocardial infarcted area, improve blood supply, and enhance the survival rate of skeletal muscle satellite cells after transplantation, rhEPO and akeletal muscle satellite cells have cooperation on treatment of acute myocardial infarction.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第29期5639-5644,共6页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
深圳市科技局资助项目(200602004)~~