摘要
目的探讨低氧化应激下肿瘤坏死因子α(TNF-α)过表达对牙髓干细胞(DPSCs)生物学性能的影响。方法实验分组为常氧DPSCs组、常氧空载转染组、常氧TNF-α过表达组、DPSCs 3%O_(2)组、空载DPSCs 3%O_(2)组、TNF-α过表达3%O_(2)组。牙髓干细胞复苏并培养,然后将慢病毒TNF-α转染细胞置于37℃、CO_(2)培养箱培养,ELISA法检测各组细胞TNF-α的表达水平,RT-PCR检测TNF-α过表达氧化应激作用下的表达,Real-time PCR、Western blotting检测氧化应激作用下人DPSCs碱性磷酸酶(ALP)、牙本质涎磷蛋白(DSPP)表达和MTT检测各组细胞TNF-α转染后细胞增殖情况。结果Lentivirus-TNF-α转染细胞10 h后,即可在倒置荧光显微镜下观察到绿色荧光信号,TNF-α过表达可以使低氧环境中牙髓干细胞增殖能力增强,且低氧的微环境可以增加TNF-α的表达,使DPSCs可以进一步的表达TNF-α,而且TNF-α过表达3%O_(2)时,Western blotting和Real-time PCR检测都表明DPSCs中ALP、DSPP表达差异具有显著性(P<0.05)。结论低氧微环境中DPSCs过表达TNF-α可以同时提高增殖效率和较高的分化效果。
Objective To investigate the effect of tumor necrosis factor-α(TNF-α)on biological properties of dental pulp stem cells(DPSCs)in low-oxidation stress.Methods The experimental groups were as follows:normox,normox control plasmid,normox-TNF-α,3%O_(2),control plasmid-3%O_(2)、TNF-α-3%O_(2).DPSCs were resuscitated and cultured firstly,then lentivirus TNF-αtransfected cells were cultured in a CO_(2) incubator at 37℃.The expression of TNF-αwas detected by ELISA in each group,and the expression of TNF-αunder oxidative stress was detected by RT-PCR,the expressions of alkaline phosphatase(ALP)and dentin sialophosphoprotein(DSPP)were detected by Real-time PCR and Western blotting,the proliferation of DPSCs cells transfected with TNF-αwas detected by flow cytometry and MTT.Results After lentivirus-TNF-αtransfected cells for 10 hours,the green fluorescence signal could be observed under inverted fluorescence microscope,TNF-αoverexpression could enhance the proliferation of DSPP in hypoxic environment,and hypoxia microenvironment can greatly increase the expression of TNF-α,Western blotting and Real-time PCR showed that there were significant differences in the expression of ALP and DSPP in DPSCs(P<0.05).Conclusion TNF-αoverexpression can improve the proliferation efficiency and differentiation efficiency of DPSCs in hypoxic microenvironment.
作者
李伟
杨赣军
彭建安
胡红梅
周星辰
LI Wei;YANG Gan-jun;PENG Jian-an;HU Hong-mei;ZHOU Xing-chen(Oral Department of Medical College,Jinggangshan University;Department of Histology and Anatomy of Medical college,Jinggangshan University,Jiangxi Ji'an 343000,China)
出处
《解剖学报》
CAS
CSCD
北大核心
2022年第1期66-74,共9页
Acta Anatomica Sinica
基金
国家自然科学基金(81660188)
江西省自然科学基金(20202BAB206037)。
关键词
低氧化
肿瘤坏死因子-Α
牙髓干细胞
免疫印迹法
人
Micro oxidation
Tumor necrosis factor-α
Dental pulp stem cells
Western blotting
Human
