肌内注射人脐带间充质干细胞对扩张型心肌病大鼠心脏功能及心肌微血管再生的影响  

Effect of human umbilical cord mesenchymal stem cells by intramuscular injection on cardiac function and microvascular regeneration in rats with dilated cardiomyopathy

作　　者：王哲哲 胡小龙 毛成刚[3] 李荣[4] 李自普 Wang Zhezhe;Hu Xiaolong;Mao Chenggang;Li Rong;Li Zipu(Qingdao University School of Medicine,Qingdao 266071,Shandong Province,China;Department of Emergency,Jiangxi Provincial Children′s Hospital,Nanchang 330006,China;Department of Cardiovascular and Renal Immunology,Affiliated Hospital of Qingdao University,Qingdao 266003,Shandong Province,China;Department of Echocardiography,Affiliated Hospital of Qingdao University,Qingdao 266003,Shandong Province,China;Heart Center,Qingdao Women and Children′s Hospital,Qingdao 266034,Shandong Province,China)

机构地区：[1]青岛大学医学部,266071 [2]江西省儿童医院急诊科,南昌330006 [3]青岛大学附属医院心肾免疫科,266003 [4]青岛大学附属医院心脏超声科,266003 [5]青岛市妇女儿童医院心脏中心,266034

出　　处：《中华实用儿科临床杂志》2018年第15期1170-1175,共6页Chinese Journal of Applied Clinical Pediatrics

基　　金：青岛市科技支撑计划（14-2-3-16-nsh）

摘　　要：目的探讨肌内注射人脐带间充质干细胞（hUCMSCs）对多柔比星诱导扩张型心肌病（DCM）大鼠心脏功能与心肌微血管再生的影响。方法将100只SD大鼠按随机数字表法分为正常组（15只）与DCM组（85只），DCM组大鼠每周腹腔注射多柔比星（2.0 mg/kg）1次，连续8周。建模成功后的大鼠随机分为DMEM细胞培养基组（DMEM组，20只）、低剂量组（20只）和高剂量组（20只），分别经四肢肌肉注射给予2 mL细胞培养基、2 mL hUCMSCs细胞悬液（1×10^6个细胞）和2 mL hUCMSCs细胞悬液（10×10^6个细胞）。应用超声心动图和血浆B型尿钠肽（BNP）水平评估正常组和DCM组大鼠心脏功能，hUCMSCs干预4周后超声检测正常组和各剂量组大鼠心功能变化；腹腔注射停止1周后，取正常组与DCM组大鼠心室肌进行HE及Masson染色，观察各组大鼠心肌病理学改变；干细胞干预停止后1周，取大鼠心肌组织，Western blot检测各组大鼠心肌血管内皮生长因子（VEGF）表达情况；免疫组织化学标记CD34，计算微血管密度；透射电镜检测心肌微血管结构改变。结果DCM组左心室射血分数（LVEF）、左心室短轴缩短率（LVFS）[（66.17±3.54）%和（31.33±3.20）%]显著低于正常组[（77.25±3.40）%和（41.00±2.94）%]，差异均有统计学意义（t＝10.620、10.328，均P〈0.05）；HE染色显示造模后的大鼠心肌细胞水肿，细胞排列紊乱，有炎症病变；Masson染色可观察到DCM组大鼠心肌纤维沉积增多。hUCMSCs治疗4周后，低剂量组和高剂量组大鼠LVEF[（72.27±2.44）%和（70.92±2.68）%]明显高于DMEM组（62.89±2.54）%，差异均有统计学意义（t＝2.145、2.131，均P〈0.05）；LVFS[（34.96±2.08）%和（33.49±2.19）%]亦明显高于DMEM组（30.98±2.22）%，差异均有统计学意义（t＝2.491、4.086，均P〈0.05）；低剂量组和高剂量组血浆BNP水平[（202.68±20.38） ng/L和（193.62±15.41） ng/ObjectiveTo evaluate the effect of human umbilical cord mesenchymal stem cells （hUCMSCs） treatment through intramuscular administration on the heart function and angiogenesis of the myocardium in dilated car-diomyopathy （DCM） rats induced by Adriamycin（ADR）.MethodsOne hundred male SD rats were randomly divided into the normal group and the DCM group.Rats in the DCM group were treated with ADR by intraperitoneal injection of 2.0 mg/kg dose per week for 8 weeks in order to induce DCM.Sixty modeled surviving rats with DCM were randomly divided equally into 3 groups, and they were treated with hUCMSCs or DMEM by intramuscular injection.Rats in the DMEM group （20 cases） received intramuscular infusion 2 mL DMEM alone; rats in the low-dose group （20 cases） underwent intramuscular infusion of 1×10^6 hUCMSCs/2 mL in DMEM; rats in the high dose group （20 cases） underwent intramuscular infusion of 10×10^6 hUCMSCs/2 mL in DMEM.Echocardiography and plasma brain natriuretic peptide（BNP） were used to assess cardiac function in modeled rats.The morphological changes in myocardial cells were observed by using HE and Masson staining after ADR injection stopped for one week.Four weeks after administration of hUCMSCs, echocardiography was performed to evaluate the cardiac function, and plasma BNP level was detected by enzyme immunoassay kit.Western blot was used to analyze the expression of vascular endothelial growth factor（VEGF） in myocardium of rats in each group.Myocardial microvessel density was detected by using anti-CD34 monoclonal antibody and transmission electron microscopy （TEM） were performed to observe the ultrastructure of microvessel.ResultsLeft ventricular ejection （LVEF） and left ventricular fractional shortening （LVFS） in the DCM groups[（ 66.17±3.54）%, （31.33±3.20）%] were significantly decreased compared to those in the normal group[（77.25±3.40）%, （41.00±2.94）%], and the differences were statistically significant（t=10.620, 10.328, all P〈0.05）;

关 键 词：扩张型心肌病 心功能 微血管再生 干细胞治疗 肌肉注射 大鼠 

分 类 号：R725.4[医药卫生—儿科]