摘要
目的:研究对比脂肪干细胞(Adipose-derived stem cells,ADSCs)、血管内皮生长因子(Vascular endothelial growth factor,VEGF)和粒细胞-巨噬细胞集落刺激因子(Granulocyte-macrophage colony stimulating factor,GM-CSF)对放射后成纤维细胞(Fibroblasts,Fb)修复的影响机制。方法:使用剂量为8 Gy的X线放射源,对大鼠的成纤维细胞进行单次X线照射,建立ADSCs与放射后成纤维细胞的共培养模型。分别设置单纯放射后成纤维细胞为Fb2组,ADSCs与放射后成纤维细胞共培养为Fb3组,VEGF+放射后成纤维细胞为VEGF组,GM-CSF+放射后成纤维细胞为GM-CSF组。通过VEGF、GM-CSF、ADSCs分别干预放射损伤的成纤维细胞,观察细胞凋亡、增殖、细胞周期及Western-Blot检测信号通路激活情况验证结果。结果:VEGF、GM-CSF、ADSCs分别作用于放射损伤的成纤维细胞,可以促进细胞增殖、抑制细胞凋亡,加快细胞周期由G2/M向G1/S进展,与照射组相比差异有统计学意义(P<0.05),其程度强弱依次为ADSCs、GM-CSF、VEGF。VEGF组和GM-CSF组p-Akt蛋白和p-Erk蛋白表达较照射组上调,差异有显著统计学意义(P<0.01);相同浓度ADSCs、GM-CSF和VEGF促进p-Akt蛋白和p-Erk蛋白表达,作用程度强弱依次为ADSCs、GM-CSF、VEGF,差异有显著统计学意义(P<0.01)。磷脂酰肌醇-3-激酶/蛋白激酶B(Phosphatidylinositol-3-kinase/protein kinase B,PI3K-Akt)通路和丝裂原激活蛋白激酶(Mitogen activated protein kinase,MAPK)通路可能发挥着重要作用,比如在VEGF和GM-CSF作用于ADSCs治疗放射损伤后成纤维细胞过程中。ADSCs修复放射损伤后成纤维细胞的效果明显好于VEGF和GM-CSF单独修复放射损伤后成纤维细胞,表明可能还有其他的潜在作用因子。结论:ADSCs、VEGF和GM-CSF干预的放射损伤后成纤维细胞主要通过增强细胞增殖、抑制细胞凋亡及加快细胞周期由G2/M向G1/S发展。
Objective To discuss the effect mechanism of adipose-derived stem cells(ADSCs)on irradiation fibroblasts(Fb)through vascular endothelial growth factor(VEGF)and granulocyte-macrophage colony stimulating factor(GM-CSF).Methods Rats Fb were irradiated with a dose of 8 Gy X-ray source,and the co-culture of ADSCs and post-irradiated Fb was established.ADSCs were co-cultured with irradiation Fb as ADSC group,VEGF+irradiation Fb was set as VEGF group,and GM-CSF+after-radiation Fb was set as GM-CSF group.VEGF,GM-CSF and ADSCs were used to intervene in radiation-injured fibroblasts,respectively,and cell apoptosis,proliferation,cell cycle,and activation of signaling pathways by Western-Blot were analyzed to verify the results.Results VEGF,GM-CSF and ADSCs intervened in radiation-injured fibroblasts,respectively,and the degrees of promoting cell proliferation,anti-apoptosis and accelerating cell cycle.Compared with the irradiation group,the differences were statistically significant(P<0.05),and its degree was ADSCs,GM-CSF,and VEGF in order.The expressions of p-Akt protein and p-Erk protein in the VEGF group and GM-CSF group were up-regulated compared with the irradiation group,and the difference was statistically significant(P<0.01).The expression of p-Erk protein and the degree of action were ADSCs,GM-CSF,and VEGF in order,and the difference was statistically significant(P<0.01).The PI3K-Akt(phosphatidylinositol-3-kinase/protein kinase B)pathway and the MAPK(mitogen-activated protein kinase)pathway may play an important role,such as the role of VEGF and GM-CSF in the treatment of ADSCs on irradiation fibroblasts.The effect of VEGF and GM-CSF is much weaker than that of ADSCs,which implies that they may only be part of the cytokines during the progress of radiation-induced skin repairment.Conclusion The radiation-injured fibroblasts treated with VEGF and GM-CSF demonstrated enhanced proliferation activity,reduced apoptosis,and accelerated cell cycle progressed from G2/M to G1/S.
作者
尹小芳
秦岭
杨超
阎海萍
栗颖利
李敏
YIN Xiaofang;QIN Ling;YANG Chao;YAN Haiping;LI Yingli;LI Min(Department of Nuclear Medicine of the 960th Hospital of the PLA,Jinan 250031,Shandong,China;Department of Burn and Plastic Surgery,960th Hospital of the People's Liberation Army,Jinan 250031,Shandong,China;People's Liberation Army Navy Special Medical Center Plastic Surgery,Shanghai 200000,China;Jinan Military Region 11th Retired Cadre Rest Center,Jinan 250000,Shandong,China)
出处
《中国美容医学》
CAS
2023年第8期29-34,共6页
Chinese Journal of Aesthetic Medicine
基金
济南市临床医学科技创新计划(编号:202019015)
国家自然科学基金项目(编号:81301644)。
关键词
脂肪干细胞
成纤维细胞
辐射
粒细胞-巨噬细胞集落刺激因子
血管内皮生长因子
adipose-derived stem cells
fibroblasts
radiation
Granulocyte-macrophage colony-stimulating factor
Vascular endothelial growth factor
