牙髓干细胞通过调节氧化应激修复肺微血管内皮细胞低氧损伤  

Mesenchymal stem cells inhibit hypoxic damage to rat pulmonary microvascular endothelial cells by regulating oxidative stress

作　　者：毛壮 李雪 王唱垚 吕琳 曹虎 何智超 余祖胤 王华 MAO Zhuang;LI Xue;WANG Changyao;LYU Lin;CAO Hu;HE Zhichao;YU Zuyin;WANG Hua(Academy of Military Medical sciences,Beijing 100850,China;School of Life Sciences,AnHui University,Hefei 230032,China;School of Life Sciences,Hebei University,Baoding 071002,China)

机构地区：[1]军事医学研究院,北京100850 [2]安徽医科大学生命科学学院,安徽合肥230032 [3]河北大学生命科学学院,河北保定071002

出　　处：《中国药理学与毒理学杂志》2024年第7期504-510,共7页Chinese Journal of Pharmacology and Toxicology

摘　　要：目的探讨牙髓干细胞(DPSC)对肺微血管内皮细胞(PMVEC)低氧损伤的修复作用及机制。方法①建立PMVEC低氧损伤模型,用氯化钴0(细胞对照),10,25,50和100μmol·L^(-1)处理PMVEC 72 h,CCK-8法检测细胞存活率,Western印迹法检测低氧诱导因子1α(HIF-1α)、闭锁小带蛋白1(ZO-1)和闭合蛋白(OCLN)的蛋白表达水平。②设细胞对照组、模型组和模型+DPSC组,免疫荧光法检测细胞活性氧(ROS)水平;Western印迹法检测ZO-1和OCLN蛋白水平。③设模型组、模型+DPSC组和模型+DPSC敲低超氧化物歧化酶1(SOD1)组,Western印迹法检测ZO-1和OCLN蛋白水平。结果①与细胞对照组相比,氯化钴100μmol·L^(-1)处理PMVEC后细胞存活率>80%,HIF-1α蛋白水平升高(P<0.05),ZO-1和OCLN蛋白水平降低(P<0.01),PMVEC低氧损伤模型构建成功。②与细胞对照组相比,模型组ZO-1和OCLN蛋白水平降低(P<0.01),ROS水平升高(P<0.01);与模型组相比,ZO-1和OCLN蛋白水平升高(P<0.05),模型+DPSC组ROS水平降低(P<0.01);③与模型组相比,模型+DPSC组ZO-1和OCLN蛋白水平升高(P<0.01);与模型+DPSC组相比,模型+DPSC敲低SOD1组ZO-1和OCLN蛋白水平降低(P<0.01)。结论DPSC通过调节氧化应激修复PMVEC低氧损伤。OBJECTIVE To explore the role and mechanism of dental pulp stem cells(DPSCs)in repairing hypoxic injury to rats pulmonary microvascular endothelial cells(PMVECs).METHODS①PMVECs were treated with cobalt chloride at 0,10,25,50 and 100μmol·L^(-1) for 72 h.CCK-8 was used to detect the cell viability,and the protein levels of hypoxia-inducible factor 1α(HIF-1α),zona occludens small-band protein 1(ZO-1),and occludin(OCLN)were detected by Western blotting.②There was a cell control group,model group,and model+DPSCs group,and the levels of reactive oxygen species(ROS)was detected by immunofluorescence staining after at 24 and 48 h of action.The levels of ZO-1 and OCLN proteins were detected by Western blotting.③A cell control group,model group,model+DPSC group and model+DPSC cell knockdown superoxide dismutase 1(SOD1)group were set up.The mRNA level of SOD1 was detected by real-time fluorescence quantitative PCR 24 and 48 h later,while the protein levels of ZO-1 and OCLN were detected by Western blotting.RESULTS①Compared with the cell control group,72 h of cobalt chloride 100μmol·L^(-1) treatment of PMVECs resulted in a cell survival rate above 80%,a significant increase in the level of HIF-1α protein(P<0.05),a significant decrease in the levels of ZO-1 and OCLN proteins(P<0.01),and establishment of a model of hypoxic injury in PMVECs.②Compared with the cell control group,the ROS level was significantly higher in the model group(P<0.01).Compared with the model group,the ROS level was significantly lower in the model+DPSCs group(P<0.01),while the levels of ZO-1 and OCLN proteins were significantly higher in the model+DPSCs group(P<0.05).③Compared with the DPSC group,ZO-1 and OCLN expressions were significantly decreased after knockdown of SOD1 in DPSCs(P<0.05,P<0.01).CONCLUSIONS DPSCs can repair hypoxic injury to PMVECs,and the anti-oxidative stress capacity of DPSCs plays an important role in hypoxic injury repair of PMVECs.

关 键 词：牙髓干细胞 氧化应激 低氧损伤 

分 类 号：R966[医药卫生—药理学]