α2-巨球蛋白减轻X射线所致人骨髓间充质干细胞成骨分化障碍  

α2-macroglobulin alleviates X-ray induced obstacle on osteogenic differentiation of human bone marrow mesenchymal stem cells

作　　者：刘仰 孔祥波[2] 李洁 陈雪英[1] 温创宇[3] 房思炼[1] 

机构地区：[1]中山大学附属第六医院口腔颌面外科,广东广州510655 [2]中山大学孙逸仙纪念医院口腔科,广东广州510120 [3]中山大学附属第六医院广东省胃肠病学研究所,广东广州510655

出　　处：《中国病理生理杂志》2017年第11期2032-2037,共6页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目(No.2015A030313064);广东省科技计划(No.2014A020212127)

摘　　要：目的:初步探讨α2-巨球蛋白(α2M)对X射线导致的人骨髓间充质干细胞(h BMMSCs)成骨分化障碍的作用。方法:体外培养h BMMSCs,取第4代细胞经8 Gy X射线照射后,进行成骨诱导并以0.5和1.0 g/Lα2M分别作用于照射后的h BMMSCs。成骨诱导的第7天检测碱性磷酸酶(ALP)活性,RT-q PCR法检测Runt相关转录因子2(RUNX2)的mRNA表达;成骨诱导的第14天采用Western blot检测骨甘氨酸(OGN)的蛋白表达;成骨诱导的第21天采用茜素红染色法检测钙结节的形成情况。此外,正常培养的h BMMSCs于8 Gy X射线照射后加入α2M作用24 h,提取细胞检测超氧化物歧化酶(SOD)活力,Western blot检测含锰超氧化物歧化酶(Mn SOD)的蛋白表达。结果:h BMMSCs经8 Gy X射线照射后给予0.5和1.0 g/Lα2M处理,与未加入α2M的单独照射组比较,ALP活性、RUNX2的mRNA表达、OGN和Mn SOD的蛋白表达及SOD活力均升高,钙结节形成增多。结论:α2M可明显提高放射损伤后h BMMSCs的成骨分化能力,上调SOD活力和Mn SOD的蛋白表达水平。AIM：To evaluate the effect of α2-macroglobulin（α2 M） against X-ray induced obstacle on osteogenic differentiation of human bone marrow mesenchymal stem cells（h BMMSCs）.METHODS：h BMMSCs were cultured in vitro.The 4 th generation of h BMMSCs was irradiated with 8 Gy X-ray,then induced osteogenic differentiation and treated with different concentrations of α2 M（0.5 and 1.0 g/L）.The alkaline phosphatase（ALP） activity and the mRNA expression of runt-related transcription factor-2（RUNX2） were detected on day 7 after osteogenic induction.The protein expression of osteoglycin（OGN） was evaluated by Western blot on day 14 after osteogenic induction.The formation of calcium nodules was detected by alizarin red staining on day 21 after osteogenic induction.The activity of superoxide dismutase（SOD） and the protein expression of Mn SOD of irradiated h BMMSCs with 8 Gy X-ray were determined at 24 h after α2 M treatment.RESULTS：Compared with 8 Gy X-ray group,the activity of ALP,the mRNA expression of RUNX2,the protein expression of OGN and Mn SOD,as well as SOD activity were higher than those in the h BMMSCs treated with α2 M at0.5 and 1.0 g/L after 8 Gy X-ray irradiation,and the calcium nodules were also increased.CONCLUSION：α2 M significantly improves the osteogenic differentiation ability,the SOD activity and Mn SOD protein expression of h BMMSCs after radiation injury.

关 键 词：放射损伤 Α2-巨球蛋白 人骨髓间充质干细胞 成骨分化 

分 类 号：R818[医药卫生—放射医学] R363[医药卫生—临床医学]