低氧与过氧化氢预处理调控间充质干细胞归巢能力的比较研究OACSTPCD
A comparative study of regulation of the homing ability of mesenchymal stem cells by hypoxia and hydrogen peroxide pretreatment
目的 为提升间充质干细胞(MSC)归巢能力,通过比较不同氧浓度及过氧化氢(H2O2)环境下MSC中趋化因子(C-X-C基序)受体4(CXCR4)表达量,以确定促进CXCR4上调的最优条件.方法 分别在0.1%、1%、3%O2及H2O2(50 μmol/L)条件下培养MSC,并于不同时间点(3、6、12、24 h)收集细胞,通过Western印迹、实时定量PCR(qPCR)、免疫荧光染色方法测定MSC中CXCR4的表达量;用CCK-8实验、划痕实验、Transwell迁移实验检测MSC经不同处理后细胞活力及趋化能力的变化.结果 经低氧及H2O2处理后,发现MSC中CXCR4均有一定程度的上调,结合蛋白和mRNA的表达情况,在3%O2条件下处理时间为6~12 h时有相对较高的表达水平,H2O2则在培养6 h之后CXCR4表达有显著上调;在低氧与H2O2处理后的细胞活力变化中发现,与对照组相比,3%O2处理后MSC细胞活力显著上调;在迁移能力检测中,3%O2与H2O2预处理均可以提升MSC的趋化迁移能力.结论 3%O2、H2O2预处理均可以上调MSC中CXCR4表达及提高MSC的迁移能力,且3%O2处理效果更佳,因此3%O2处理条件为上调MSC趋化能力的最佳选择.
Objective To determine the optimal conditions for CXCR4 upregulation by comparing the expression levels of chemokine(C-X-C motif)receptor 4(CXCR4)in MSCs cultured with varying concentrations of oxygen and hydrogen peroxide(H2O2).Methods MSCs were cultured with 0.1%,1%,or 3%O2 and 50 μmol/L H2O2 for different lengths of time(3,6,12,and 24 h).The mRNA and protein expressions of CXCR4 in MSCs were measured by real-time quantitative PCR(qPCR),Western blotting,and immunofluorescence staining.The viability and chemotactic ability of MSCs were measured using CCK-8,wound-healing and Transwell migration assays.Results Both hypoxia and H2O2 treatment were found to upregulate MSC expressions of CXCR4 to some extent.The mRNA and protein levels of CXCR4 were higher after 6-12 h of culture of MSCs with 3%O2,and significantly higher when treated with H2O2 for 6 h.Cell viability was significantly increased after culture with 3%O2 compared with the control group and both 3%O2 and H2O2 pretreatment could enhance chemotactic migration in MSCs.Conclusion Culture with 3%O2 and H2O2 pretreatment can upregulate CXCR4 expressions in MSCs and enhance migration in cells,with superior effects observed with 3%O2.Therefore,treatment with 3%O2 represents the best choice for upregulating the chemotactic ability of MSCs.
黄一琛;姜明霞;马文静;陈巧汨;白睿;修冰水;宋梦文;牛洁;刘志强
南华大学衡阳医学院,军事科学院军事医学研究院研究生协作培养基地,湖南衡阳 421001潍坊医学院附属医院,烟台市莱阳中心医院,山东烟台 265299安徽医科大学基础医学院,合肥 230032解放军总医院第六医学中心心血管病医学部,北京 100853军事科学院军事医学研究院,北京 100850首都医科大学石景山教学医院,北京石景山医院,北京 100040南华大学衡阳医学院,军事科学院军事医学研究院研究生协作培养基地,湖南衡阳 421001||军事科学院军事医学研究院,北京 100850
基础医学
间充质干细胞战创伤趋化因子(C-X-C基序)受体4归巢低氧H2O2
mesenchymal stem cellswar woundCXCR4hominghypoxiaH2O2
《军事医学》 2024 (009)
656-663 / 8
国家自然科学基金项目(82200295,81971168)
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