| Purposes:1.To investigate the effects of ROS level in local tissues during the occurrence and development of radioactive stomatitis of mice.2.To study the effects of GMSCs improve the ROS level in tissues of radioactive stomatitis of mice in vivo.Methods:1.Isolation and culture of GMSCs of C57/BL mice.2.Select C57/BL mice of 6-8 weeks and randomly divided into 6 groups: Normal group (no radiation); Treatment experimental group (25Gy radiation dose was given to each mouse by electron linear accelerator.); Treatment control group (injection of 106 GMSCs by tail veins 7 days after radiation) Prevention experimental group (injection of cell-free DMEM medium by tail veins 3 days after radiation) Prevention control group (injection of 106 GMSCs by tail veins 30 min before radiation) Control group (injection of cell-free DMEM medium by tail veins 30 min before radiation)3.The establish animal models of C57/BL mice of radioactive stomatitis:using electron linear accelerator to build radiation mice model, in the dose of 25Gy each time, and then observe histological sections with optical microscope.4.Detection of SOD and MDA level in lips and tongues of mice:4.1 Cutting perioral tissues of Normal group and Control groups 3 days,7 days and 10 days after radiation, and determine SOD and MDA level by colorimetric analysis.4.2 Cutting perioral tissues of Treatment experimental group, Treatment control group, Prevention experimental group and Prevention control group respectively 3 days,7 days and 10 days after injection, and determine SOD activity and MDA content by colorimetric analysis.Results:1.Separation and culture of GMSCs of C57/BL mice2.Successfully established mice model of radioactive stomatitis.:The histopathological changes were observed under microscope after radiation, sections showed that epithelial continuity interruption, inflammatory cell infiltration in mucosa and telangiectasia, and these changes deteriorated over time3.SOD and MDA level of tissues:1) SOD activity of lip tissue:Radiation group is increased significantly than Normal group (p<0.05), no significant difference between Prevention experimental group and Prevention control group (p>0.05), Treatment experimental group is increased significantly compared with Treatment control group (p<0.05) after 3 days; Radiation group is decreased significantly than Normal group (p<0.05), Prevention experimental group and Treatment experimental group were increased significantly respectively compared with their control group after 7 days and 10 days.(p<0.05).2) MDA content of lip tissue:no significant difference of MDA content between Radiation group and Normal group (p>0.05), and no significant difference between Prevention experimental group and Prevention control group (p>0.05), Treatment experimental group is decreased significantly compared with Treatment control group (p<0.05) after 3 days; Radiation group is increased significantly than Normal group (p<0.05), Prevention experimental group and Treatment experimental group were decreased significantly respectively compared with their control group after 7 days and 10 days3) SOD activity of tongue tissue:Radiation group is increased significantly than Normal group (p<0.05), no significant difference between Prevention experimental group and Prevention control group (p>0.05), Treatment experimental group is increased significantly compared with Treatment control group (p<0.05) after 3 days; Radiation group is decreased significantly than Normal group (p<0.05), Prevention experimental group and Treatment experimental group were increased significantly respectively compared with their control group after 7 days and 10 days. (p<0.05)4) MDA content of tongue tissue:no significant difference between Radiation group and Normal group (p>0.05), and no significant difference between Prevention experimental group and Prevention control group respectively compared with their control group (p>0.05) after 3 days, Radiation group is increased significantly than Normal group (p<0.05), Prevention experimental group and Treatment experimental group were decreased significantly respectively compared with their control group after 7 days and 10 days (p<0.05)Conclusion:1. The histopathological changes were observed under microscope after radiation, sections showed that epithelial continuity interruption, inflammatory cell infiltration in mucosa and telangiectasia, and these changes deteriorated over time2. The comparison of SOD activity and MDA content in tissues indicate that in experimental mice models, SOD activity in perioral tissue declined while the content of MDA increased during the radioactive oral mucositis process, and the weakening of activity of SOD in tissues are involved in the occurrence and development of radioactive stomatitis.3. SOD activity in tissue increased and MDA content decreased when using GMSCs as prevention or treatment methods, indicates that GMSCs play a role in scavenging oxygen free radicals, improve oxygen free radical levels in local tissue, relieve inflammatory state of oral mucosa, and have effects on prevention and treatment of radioactive stomatitis. |
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