骨髓间充质干细胞外泌体对雪旺细胞的增殖迁移、BDNF和VEGF分泌能力影响观察OACSTPCD
Effects of bone marrow mesenchymal stem cell exosomes on proliferation,migration,BDNF,and VEGF secretion of Schwann cells
目的 观察骨髓间充质干细胞(BMSCs)分泌的外泌体对雪旺细胞增殖迁移、脑源性神经营养因子(BDNF)和血管内皮生长因子(VEGF)分泌能力影响.方法 BMSCs分泌的外泌体提取自3周龄SD大鼠双侧股骨骨髓,雪旺细胞提取自3日龄SD大鼠双侧坐骨神经.将雪旺细胞分为对照组、BMSCs组,对照组采用雪旺细胞培养基培养,BMSCs组使用含有100 μg/mL BMSCs外泌体的雪旺细胞培养基培养.两组雪旺细胞继续培养24、48、72 h时,采用CCK-8法观察两组雪旺细胞增殖能力(以OD值表示).两组雪旺细胞继续培养24、48 h时,采用划痕实验观察两组雪旺细胞迁移能力(以迁移相对面积比表示).两组雪旺细胞继续培养72 h时,采用ELISA法观察两组雪旺细胞BDNF、VEGF分泌能力(以细胞上清液中BDNF、VEGF含量表示).结果 对照组雪旺细胞培养24、48、72 h时的OD值分别为1.06±0.04、1.22±0.06、1.42±0.08,BMSCs组分别为1.46±0.11、1.70±0.10、1.94±0.19,两组相比,P均<0.05.对照组雪旺细胞培养24、48 h时的迁移相对面积比分别为26.43%±4.75%、45.44%±3.41%,BMSCs组分别为46.29%±3.88%、65.04%±2.85%,两组相比,P均<0.05.对照组雪旺细胞培养72 h时的细胞上清液中BDNF含量为(72.48±7.25)pg/mL,BMSCs组雪旺细胞上清液中BDNF含量为(109.70±6.38)pg/mL,两组相比,P<0.05.对照组雪旺细胞培养72 h时细胞上清液中VEGF含量为(176.00±6.90)pg/mL,BMSCs组雪旺细胞上清液中VEGF含量为(247.10±16.37)pg/mL,两组相比,P<0.05.结论 BMSCs分泌的外泌体可以促进雪旺细胞的增殖迁移和BDNF、VEGF分泌能力,有助于神经再生.
Objective To investigate the effects of exosomes secreted by bone marrow mesenchymal stem cells(BM-SCs)on the proliferation,migration,brain-derived neurotrophic factor(BDNF)and vascular endothelial growth factor(VEGF)secretion of Schwann cells.Methods Exosomes secreted by BMSCs were extracted from bilateral femoral bone marrow of 3-week-old SD rats,and Schwann cells were extracted from bilateral sciatic nerves of 3-day-old SD rats.Schwann cells were divided into the control group and BMSCs group.Cells in the control group were cultured with Schwann cell medium,and cells in the BMSCs group were cultured with Schwann cell medium containing 100 μg/mL BMSCs exo-somes.When Schwann cells were cultured for 24,48 and 72 h,the proliferation of Schwann cells in the two groups was ob-served by CCK-8(expressed by OD value).When Schwann cells were cultured for 24 and 48 h,the migration abilities of Schwann cells in the two groups were observed by Scratch test(expressed by migration relative area ratio).When Schwann cells of the two groups were cultured for 72 h,the secretion capacities of BDNF and VEGF were observed by ELISA meth-od(represented by the content of BDNF and VEGF in the cell supernatant).Results The OD values of Schwann cells cultured for 24,48 and 72 h were 1.06±0.04,1.22±0.06 and 1.42±0.08,respectively,in the control group,versus 1.46±0.11,1.70±0.10 and 1.94±0.19,respectively,in the BMSCs group,with statistically significant difference(all P<0.05).The relative area ratios of Schwann cells in the control group were 26.43%±4.75%and 45.44%±3.41%at 24 and 48 h,respectively,and those in the BMSCs group were 46.29%±3.88%and 65.04%±2.85%,respectively,with statistically significant difference(all P<0.05).The content of BDNF in the supernatant of Schwann cells cultured for 72 h in the control group was(72.48±7.25)pg/mL,and that in the supernatant of Schwann cells in the BMSCs group was(109.70±6.38)pg/mL,with statistically significant difference(P<0.05).Conclusion Exosomes secreted by BMSCs can promote the proliferation,migration and the secretion of BDNF and VEGF of Schwann cells,thus contributing to nerve regeneration.
李慈;赵闯
山东第一医科大学附属省立医院创伤中心,济南 250021
临床医学
外泌体间充质干细胞骨髓间充质干细胞雪旺细胞周围神经损伤神经再生
exosomesmesenchymal stem cellsbone marrow mesenchymal stem cellsSchwann cellsperipher-al nerve injurynerve regeneration
《山东医药》 2024 (033)
11-14 / 4
山东省自然科学基金项目(ZR2023QH298);山东第一医科大学附属省立医院科研孵育基金立项项目(2022FY035).
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