自体骨髓间充质干细胞移植治疗杜氏型肌营养不良血清酶学变化对照研究

Curative effect analysis of autologous bone marrow mesenchymal stem cells transplantation in Duchenne muscular dystrophy

目的观察骨髓间充质干细胞（BMSC）移植治疗杜氏型肌营养不良（DMD）治疗前后血清酶学变化及临床效果。方法选择2007年6月至2009年6月解放军第四六三医院515例DMD患者,其中男性502例,女性13例;年龄1～20岁,平均年龄13.3岁。阳性家族史121例。经解放军第四六三医院伦理委员会同意,并征求患者本人及家人签字同意后,进行自体BMSC移植治疗,设自身治疗前后血清酶学变化对照方法进行疗效观察,入组患者均皮下注射粒细胞集落刺激因子,动员4d后采集骨髓,Percoll梯度离心,提取的单个核细胞总量为2.0×109,培养骨髓单个核细胞7-10d,最终得到BMSC总数4.0×108-1.5×109/mL。应用四肢肌内注射的方式进行BMSC移植。结果 BMSC移植治疗12个月肌力增加者占患者总数的79.2%。临床升级人数占27.5%。血清肌酸激酶（CK）降低率81.0%,血清乳酸脱氢酶（LDH）降低率52.8%。结论 BMSC移植治疗DMD,对肌肉组织有一定的修复作用,使DMD患者的运动功能得到改善,肌力有所提高,血清酶学较移植前下降。BMSC移植无不良反应,患者依从性好,是治疗DMD的新手段。

Objective To discuss the clinical effect of autologous bone marrow mesenchymal stem cells（BMSC） transplantation in patients with Duchenne muscular dystrophy（DMD）.Methods A total of 515 DMD patients from June 2007 to June 2009 were enrolled,male 502,female 13,aged 1-20 years old,mean age 13.3 years old,of 121 patients with positive family history.All of them performed a whole year treatment with autologous BMSC transplantation from March 2008 to March 2009.The parameters before and after treatment were compared,the granulocyte-colony stimulating factor（G-CSF） was infused into all patients,and bone marrow was collected after 4 days of mobilization.Bone marrow mononuclear cells（MNC） were isolated by Percoll gradient centrifugation.The MNC were cultured for 7-10 days and obtained the amount of BMSC 4.0 × 108 ~ 1.5 × 109.The BMSC were transplanted into all the limbs by intramuscular injection.Results One year after transplantation,increased rate of patient muscle force was 79.2 %,and 27.5 % of the patients were upgraded by the transplantation.The decreased rates of serum creatine kinase（CK） and lactic acid dehydrogenase（LDH） were respectively 81.0 % and 52.8 %.Conclusion It is demonstrated that transplantation of autologous BMSC could improve motor function and increase muscle force of DMD patient.It could decrease the level of CK and LDH in blood serum,so transplantation of autologous BMSC is a new way to treat with DMD for safety and without adverse reactions.