双特异抗体增效骨髓间充质干细胞干预心肌纤维化

Effect of bone marrow mesenchymal stem cells transplantation on fibroid myocardium with the aid of bispecific antibody

目的:探讨在双特异抗体(Bispecificantibody,BiAb)的辅助下,骨髓间充质干细胞(Bonemarrowmesenchymalstemcells,BMSCs)移植干预心肌纤维化的效果。方法:以anti-CD29(能识别间充质干细胞)和抗肌凝蛋白轻链抗体(Anti-myosin light chainantibody,AMLCA)(能特异性结合损伤心肌)制备BiAb(CD29×AMLCA)。将此BiAb与雄性小鼠BMSCs经尾静脉输入异丙肾性心肌纤维化雌性小鼠(BMSCs+BiAb组)。另设单纯BMSCs组、单纯BiAb组、正常对照组、未治疗组。5周后处死小鼠,荧光定量PCR检测心肌y染色体鉴别基因(Sex-determiningregion ofY-chromosome,SRY)、基质金属蛋白酶-9(Matrixmetalloproteinas-es-9,MMP-9)、基质金属蛋白酶组织抑制剂-1(Tissue inhibitor of metalloproteinase-1,TIMP-1)的表达。天狼猩红染色比较心脏胶原含量。结果:BMSCs+BiAb组的干细胞归巢数比BMSCs组多(P<0.05)。与未治疗组相比,BMSCs+BiAb组和单纯BMCSs组的心肌MMP-9和TIMP-1表达下调,心脏胶原沉积降低(P<0.05)。与BMSCs组相比,BMSCs+BiAb组的心肌纤维化改善更为明显(P<0.05)。结论:CD29×AMLCA双特异抗体可促进小鼠骨髓间充质干细胞的归巢。BMSCs归巢后能改善缺血心肌MMP-TIMP表达,干预心肌纤维化。

Objective： To investigate the effect of bone marrow mesenchymal stem cells（BMSCs） transplantation on fibroid myocardium with the aid of bispecific antibody（BiAb）.Methods： BiAb [anti-CD29×anti-myosin light chain antibody（AMLCA）] was prepared and combined with isolated BMSCs from male mice and transfused into female mice with isoproterenol-induced myocardial fibrosis via tail vein.This study included five groups： BMSCs＋BiAb,BMSCs,BiAb,untreated,and control groups.Five weeks after treatment,expression levels of sex-determining region of Y-chromosome（SRY）,matrix metalloproteinases-9（MMP-9）,tissue inhibitor of metalloproteinase-1（TIMP-1） in myocardium were detected by fluorescent qRT-PCR.Collagen distribution was observed using sirius red staining.Results： Higher homing number of BMSCs was shown in the BMSCs＋BiAb group than that of the BMSCs group（P0.05）.Compared with the untreated group,BMSCs＋BiAb and BMSCs groups had decreased levels of MMP-9,TIMP-1 and collagen deposition（P0.05）.Decreased level of collagen content was significantly different in BMSCs＋BiAb as compared to BMSCs group（P0.05）.Conclusion： Homing rate and repairing efficacy of BMSCs improve via treatment with combination of BiAb.BMSCs can improve MMP-TIMP expression in injured myocardium and interfere with myocardial fibrosis after homing.