凋亡抑制因子5促进干细胞修复早期股骨头坏死的研究

Baculoviral inhibitor of apoptosis repeat-containing protein 5 promotes repair of early osteonecrosis of the femoral head by bone marrow mesenchymal stem cells

目的探讨凋亡抑制因子5(BIRC5)能否促进骨髓间充质干细胞(BMSCs)对早期激素性股骨头坏死(SONFH)的修复。方法采用SD大鼠分离培养BMSCs,BIRC5慢病毒转染BMSCs,完全随机分3组:空白对照、阴性病毒、BIRC5转染;采用BMSCs与异种脱抗原松质骨(XACB)构建组织工程骨移植治疗早期SONFH,实验完全随机分五组:单纯钻孔减压、XACB、XACB/BMSCs、XACB/BMSCs/绿色荧光蛋白(Lv-GFP)、XACB/BMSCs/Lv-BIRC5/GFP。小动物成像仪检测BMSCs存活情况;术后4、8、12周,微型CT(Micro-CT)评估骨坏死修复效果;采用单因素方差分析(ANOVA)分析数据。结果BIRC5转染组的BIRC5 mRNA相对表达量高于阴性病毒组及空白对照组(7.623±0.127比3.367±0.135比3.227±0.156,F=957.483,P<0.05),BIRC5转染组的BIRC5蛋白相对表达量高于阴性病毒组及空白对照组(1.263±0.038比0.393±0.015比0.417±0.031,F=850.551,P<0.05)。XACB具有良好的生物相容性,BMSCs可在XACB表面正常生长。术后第3天,XACB/BMSCs/Lv-BIRC5/GFP组荧光强度值高于XACB/BMSCs/Lv-GFP组及XACB/BMSCs组(0.874±0.066比0.536±0.054比0.557±0.081,F=23.234,P<0.05)。在术后12周时,XACB/BMSCs/Lv-BIRC5/GFP组骨小梁数目高于XACB/BMSCs/Lv-GFP组及XACB/BMSCs组[(3.873±0.176)/mm比(2.763±0.140)/mm比(2.775±0.184)/mm,F=294.717,P<0.05],XACB/BMSCs/Lv-BIRC5/GFP组骨小梁结构完整,可见成熟骨组织散在分布,骨坏死区完全修复。结论BIRC5可促进BMSCs在坏死区的存活,增强BMSCs对早期SONFH的修复。

Objective To investigate whether baculoviral inhibitor of apoptosis repeat-containing protein 5(BIRC5)can promote the repair of early steroid-induced femoral head osteonecrosis(SONFH)by bone marrow mesenchymal stem cells(BMSCs).Methods BMSCs were isolated and cultured using SD rats;BMSCs were transfected with BIRC5 lentivirus and completely randomized into three groups:blank control group,negative virus group,and BIRC5 transfection group;BMSCs and allogeneic deantigenized cancellous bone(XACB)were used to construct tissue-engineered bone grafts for early SONFH,and the experiment was completely randomized into five groups:simple drill decompressiongroup,XACBgroup,XACB/BMSCsgroup,XACB/BMSCs/green fluorescent protein(Lv-GFP)group,XACB/BMSCs/Lv-BIRC5/GFPgroup;Detection of BMSCs survival using small animal imager;Evaluation of osteonecrosis repair by micro-CT(Micro-CT)at 4,8 and 12 weeks after surgery;One-way analysis of variance(ANOVA)was used to analyze the experimental data.Results The relative expression of BIRC5 mRNA in the BIRC5 transfected group was higher than that in the negative virus group and the blank control group(7.623±0.127 vs.3.367±0.135 vs.3.227±0.156,F=957.483,P<0.05),The relative expression of BIRC5 protein in the BIRC5 transfected group was higher than that in the negative virus group and the blank control group(1.263±0.038 vs.0.393±0.015 vs.0.417±0.031,F=850.551,P<0.05).XACB has good biocompatibility and BMSCs can grow normally on the surface of XACB.The fluorescence intensity of XACB/BMSCs/Lv-BIRC5/GFP group was higher than that of XACB/BMSCs/Lv-GFP group and XACB/BMSCs group on the 3rd postoperative day(0.874±0.066 vs.0.536±0.054 vs.0.557±0.081,F=23.234,P<0.05).At 12 weeks postoperatively,the number of bone trabeculae in XACB/BMSCs/Lv-BIRC5/GFP group was higher than that in XACB/BMSCs/Lv-GFP group and XACB/BMSCs group[(3.873±0.176)/mm vs.(2.763±0.140)/mm vs.(2.775±0.184)/mm,F=294.717,P<0.05],In the XACB/BMSCs/Lv-BIRC5/GFP group,the bone trabecular structure was intact,the mature bone tissue was scattered,and the osteonecrosis area was completely repaired.Conclusion BIRC5 can promote the survival of BMSCs in osteonecrosis area and enhances the repair of early SONFH by BMSCs.