AIM:To evaluate the effect of α2-macroglobulin(α2M) against X-ray induced obstacle on osteo-genic differentiation of human bone marrow mesenchymal stem cells(hBMMSCs). METHODS:hBMMSCs were cultured in vitro. The 4th generation of hBMMSCs was irradiated with 8 Gy X-ray,then induced osteogenic differentiation and trea-ted with different concentrations of α2M(0.5 and 1.0 g/L). The alkaline phosphatase(ALP) activity and the mRNA ex-pression of runt-related transcription factor-2 (RUNX2) were detected on day 7 after osteogenic induction. The protein ex-pression of osteoglycin (OGN) was evaluated by Western blot on day 14 after osteogenic induction. The formation of calci-um nodules was detected by alizarin red staining on day 21 after osteogenic induction. The activity of superoxide dismutase (SOD) and the protein expression of MnSOD of irradiated hBMMSCs with 8 Gy X-ray were determined at 24 h after α2M treatment. RESULTS:Compared with 8 Gy X-ray group,the activity of ALP,the mRNA expression of RUNX2,the pro-tein expression of OGN and MnSOD,as well as SOD activity were higher than those in the hBMMSCs treated with α2M at 0.5 and 1.0 g/L after 8 Gy X-ray irradiation,and the calcium nodules were also increased. CONCLUSION:α2M signifi-cantly improves the osteogenic differentiation ability,the SOD activity and MnSOD protein expression of hBMMSCs after ra-diation injury.%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(hBMMSCs)成骨分化障碍的作用.方法:体外培养hBMMSCs,取第4代细胞经8GyX射线照射后,进行成骨诱导并以0.5和1.0g/L α2M分别作用于照射后的hBMMSCs.成骨诱导的第7天检测碱性磷酸酶(ALP)活性,RT-qPCR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hBMMSCs于8 Gy X射线照射后加入α2M作用24h,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(MnSOD)的蛋白表达.结果:hBMMSCs经8 Gy X射线照射后给予0.5和1.0g/L α2M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MnSOD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hBMMSCs的成骨分化能力,上调SOD活力和MnSOD的蛋白表达水平.
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