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背景:杜氏型肌营养不良的肌细胞损伤并非单一细胞受损,而是多因素缺陷导致肌细胞进行性萎缩的严重后果,患儿一经确诊即被告之无法医治。近年来国内外学者把目光转向具有修复和再生功能的干细胞,研究发现干细胞技术可以改善肌营养不良的运动功能。目的:首次采用骨髓和脐血间充质干细胞联合移植方法,首次采用静脉注射和局部2种治疗途径,观察治疗杜氏型肌营养不良患者的安全性和有效性。设计:自身前后对照。对象:2007-06/2008-09解放军第四六三医院细胞治疗中心应用骨髓和脐血间充质干细胞联合移植方案治疗杜氏型肌营养不良患者31例,男29例,女2例(双胞胎),平均年龄9.7岁,平均病程5.1年,阳性家族史11例,均经临床症状、酶学、肌电图、基因分析、肌肉活检、肌肉核磁共振检查确诊。方法:经双侧髂后上棘抽取患者自体骨髓80～150mL,采用Ficoll密度梯度离心法分离单个核细胞,并诱导分化为骨髓间充质干细胞,调整细胞悬液浓度为1×1010L-1,采用多点注射法移植于患者四肢肌肉内,移植细胞数为(1.5～6.0)×108个。选择健康足月产妇脐带血80～145mL,同法制备脐血间充质干细胞,经静脉植入(1.0～4.8)×107个细胞,1次/周,共3次。主要观察指标:干细胞移植前后患者日常生活活动能力评价,临床分级评价,以及酶学、肌电图、肌肉MRI变化。结果:31例患者均完成15个月随访,中途无脱落。与移植前比较,干细胞移植后3,6,9,12,15个月,26例(84.7%)动作灵活度增强,运动功能改善,日常生活活动能力评分升高(P<0.05);干细胞移植后6个月,血清肌酸肌酶、乳酸脱氢酶水平降低11例,升高8例;5例复查肌电图,波幅改善2例;5例复查MRI,肌肉轮廓较前清晰2例。临床分级评定显效12例(38.7%),有效14例(45.2%),无效5例(16.1%),总有效率为83.9%,未发生不良反应及并发症。结论:骨髓和脐血间充质干细胞联合移植治疗杜氏型肌营养不良可使83.9%患者获得临床症状改善,运动功能及肌力提高,6个月后部分患者血清酶学、肌电图及肌肉MRI轻度改善,安全性好,无排斥反应,是一种治疗杜氏型肌营养不良的新手段。
BACKGROUND: Muscular dystrophy of Duchenne muscular dystrophy is not a single cell injury, but a serious consequence of multifactorial defects leading to progressive atrophy of muscle cells. Once a child is diagnosed, the accused can not heal. In recent years, scholars at home and abroad have turned their attention to stem cells that have the functions of repair and regeneration. Studies have found that stem cell technology can improve the motor function of muscular dystrophy. OBJECTIVE: To study the safety and efficacy of the combined treatment of bone marrow and umbilical cord blood mesenchymal stem cells for the first time by intravenous injection and two local routes of treatment. Design: self-control. PARTICIPANTS: 2007-06 / 2008-09 PLA Cell Center, 463 Hospital, China 31 cases of Duchenne muscular dystrophy were treated with bone marrow and umbilical cord blood mesenchymal stem cells transplantation in combination with 29 patients and 2 females (twins) , Mean age 9.7 years, average duration of disease 5.1 years, positive family history of 11 cases were diagnosed by clinical symptoms, enzymology, EMG, gene analysis, muscle biopsy, muscle MRI. Methods: The autologous bone marrow of 80-150 mL was extracted from bilateral posterior superior iliac spine. Mononuclear cells were isolated by Ficoll density gradient centrifugation and induced to differentiate into bone marrow mesenchymal stem cells. The concentration of cell suspension was adjusted to 1 × 1010 L-1, Using multi-point injection in the limbs of patients with muscle transplantation, the number of transplanted cells (1.5 ~ 6.0) × 108. Select healthy full-term maternal umbilical cord blood 80 ~ 145mL, the same method of preparation of umbilical cord blood mesenchymal stem cells, intravenous (1.0 ~ 4.8) × 107 cells, 1 / week, a total of 3 times. MAIN OUTCOME MEASURES: Evaluation of patients’ daily living activities before and after stem cell transplantation, clinical grade evaluation, and changes of enzyme, EMG and muscle MRI. Results: All 31 patients completed 15 months follow-up without any drop off. Compared with those before transplantation, 26 cases (84.7%) had more mobility, improved motor function and higher activities of daily living (P <0.05) at 3, 6, 9, 12 and 15 months after stem cell transplantation; Six months later, serum creatine kinase and lactate dehydrogenase levels were reduced in 11 cases and in 8 cases, while in 5 cases, EMG was improved and amplitude was improved in 2 cases. MRI was reviewed in 5 cases and clear in 2 cases. There were 12 cases (38.7%), 14 (45.2%) effective and 5 (16.1%) ineffective in clinical classification. The total effective rate was 83.9%. There were no adverse reactions and complications. Conclusion: The combination of bone marrow and umbilical cord blood mesenchymal stem cells in the treatment of Duchenne muscular dystrophy can improve clinical symptoms, improve motor function and muscle strength in 83.9% of patients, serum enzymes, electromyography and muscle in some patients after 6 months Mild MRI improvement, safety, and rejection are a new treatment for Duchenne muscular dystrophy.