Du Lili, Lv Runxiao, Yang Xiaoyi, Cheng Shaohang, Ma Tingxian, Xu Jing
Department of Pathophysiology, College of Basic Medical Science, China Medical University, Shenyang 110122, People's Republic of China.
Department of Spinal Surgery, Changhai Hospital, The Second Military Medical University, Shanghai 200433, People's Republic of China.
Life Sci. 2016 Mar 15;149:51-7. doi: 10.1016/j.lfs.2016.02.050. Epub 2016 Feb 15.
Scar formation after wound repair affects people's daily life. Mesenchymal stem cells were reported to have a beneficial role in attenuating the scar formation. In the present study, placenta-derived mesenchymal stem cells (PMSCs) were isolated and the effects of hypoxic conditioned medium of PMSCs on scar formation were explored.
To evaluate the effect of hypoxia on PMSCs, proliferation of PMSCs was detected by trypan blue staining and the HIF-1α level was detected by western blot. Then in vivo scar formation assay was performed and the histopathologic changes were evaluated by HE staining and levels of TGF-β1 and collagen I were detected by quantitative real-time PCR. The IL-10 level was detected by ELISA and then migration of HFF-1 cells was detected by wound healing assay after treatment with IL-10 or IL-10 antibody.
Our study showed that hypoxic conditioned medium of PMSCs reduced scar formation in vivo and inhibited the proliferation and migration of skin fibroblasts in vitro. Further mechanism study showed that, the level of IL-10 was affected by hypoxia, treatment with IL-10 mimicked the function of hypoxic conditioned medium of PMSCs and inhibition of IL-10 reversed the protective role of hypoxic conditioned medium of PMSCs. Thus, hypoxic conditioned medium of PMSCs may perform the protective role against scar formation through IL-10.
Our study reveals a possible mechanism of the protective effect of PMSCs against scar formation and provides evidence for the hypothesis that PMSCs may be a promising therapy for the treatment of wounds.
伤口修复后的瘢痕形成影响人们的日常生活。据报道，间充质干细胞在减轻瘢痕形成方面具有有益作用。在本研究中，分离了胎盘来源的间充质干细胞（PMSCs），并探讨了PMSCs缺氧条件培养基对瘢痕形成的影响。
为评估缺氧对PMSCs的影响，通过台盼蓝染色检测PMSCs的增殖，并通过蛋白质印迹法检测HIF-1α水平。然后进行体内瘢痕形成试验，通过苏木精-伊红（HE）染色评估组织病理学变化，并通过定量实时聚合酶链反应检测TGF-β1和胶原蛋白I的水平。通过酶联免疫吸附测定法检测IL-10水平，在用IL-10或IL-10抗体处理后，通过伤口愈合试验检测人包皮成纤维细胞（HFF-1细胞）的迁移。
我们的研究表明，PMSCs缺氧条件培养基在体内减少了瘢痕形成，并在体外抑制了皮肤成纤维细胞的增殖和迁移。进一步的机制研究表明，缺氧影响IL-10水平，用IL-10处理模拟了PMSCs缺氧条件培养基的功能，而抑制IL-10则逆转了PMSCs缺氧条件培养基的保护作用。因此，PMSCs缺氧条件培养基可能通过IL-10发挥抗瘢痕形成的保护作用。
我们的研究揭示了PMSCs抗瘢痕形成保护作用的一种可能机制，并为PMSCs可能是一种有前景的伤口治疗方法这一假说提供了证据。
