低氧与DMOG对BMSCs成脂分化影响的对比研究
The comparative research between hypoxia and dimethyloxalylblycine on the adipogenic differentiation of BMSCs in mouse
张磊 1邓雪春 2鲜明3
作者信息
- 1. 611135 成都,成都中医药大学附属四川省康复医院/成都中医药大学附属四川省八一康复中心脊髓损伤康复科
- 2. 610065 成都,西部战区空军医院中医科
- 3. 610041 成都,成都体育学院附属体育医院正骨科/小儿骨科
- 折叠
摘要
目的 对比低氧培养与常氧下添加二甲基乙二酰基甘氨酸(DMOG)对小鼠骨髓间充质干细胞(BMSCs)成脂分化的影响.方法 流式细胞术及细胞的成骨、成脂、成软骨三系分化对骨髓BMSCs进行鉴定;ELISA法观察低氧诱导因子-1α(HIF-1α)在对照组(21％O2)、低氧组(1％低氧培养组)及DMOG干预组(常氧+DMOG培养组)下表达情况;油红O染色法观察BMSCs内脂肪滴形成情况;分别采用Real-time PCR法和Western blot法检测各组成脂相关基因过氧化物酶增殖物激活受体γ(PPAR-γ)和CCAAT/增强子结合蛋白α(C/EBPα)的mRNA和蛋白水平.结果 流式细胞术显示骨髓单个核细胞阳性表达CD90、CD44、CD105,阴性表达CD34,且能够向成骨、成软骨、成脂三系分化;与对照组HIF-1α蛋白表达(85.00±1.87)相比,低氧组及DMOG干预组(480.40±6.91、331±7.78)明显上调(P＜0.01),其中低氧组HIF-1α蛋白较DMOG干预组HIF-1α蛋白明显上调(P＜0.01);与对照组阳性细胞数(51.80±4.15)相比,低氧组和DMOG干预组(27.80±3.11、31.20±2.59)明显减少(P＜0.05),低氧组与DMOG干预组阳性细胞数差异无统计学意义(P＞0.05).与对照组相比,低氧组与DMOG干预组PPARγ与C/EBPα的mRNA明显下调,差异具有统计学意义(P＜0.01).与低氧组相比,DMOG干预组PPAR-γ及C/EBPα的mRNA表达明显下调(P＜0.05).与对照组相比,低氧组及DMOG干预组PPAR-γ蛋白表达分别下调了 28.3％及39.8％(P＜0.01),C/EBPα的蛋白表达分别下调了 36.7％及42.5％(P＜0.01);与低氧组相比,DMOG干预组PPAR-γ及C/EBPα蛋白表达分别下降了 16％及9.1％,差异均有统计学意义(P＜0.05).结论 1％物理性低氧和常氧条件下0.5 mmol/L DMOG均能抑制BMSCs成脂分化,1％物理性低氧具有更强的抑制作用.
Abstract
Objective To investigate the effects of BMSCs on adipogenesis between that in hypoxia environment and that in normal oxygen environment with hypoxia mimetic agents dimeth-yloxalylglycine(DMOG).Methods BMSCs from the bone marrow mononuclear cells were identified by flow cytometer and osteogenic,adipogenic,chondrogenic differentiation.ELISA was employed to determine the expression of HIF-1α protein in control group(under 21％normal oxygen environment),hypoxia group(under 1％oxygen environment)and DMOG group(under 21％normal oxygen environmentplus 0.5 mmol/LDMOG).Red oil O staining was employed to observe the formation of lipid droplets in BMSCs in each group.The mRNA and protein level of PPAR-γ and C/EBPα were detected using qPCR and Western-blotting in each group.Results Flow cytometry assay showed the bone marrow mononuclear cells were positively expressed with CD90,CD44,CD 105 and negatively expressed with CD34.The cells un-derwent osteogenesis,adipogenesis,chondrogenesis.Compared with the protein expression of the HIF-1α(85.00±1.87)in control group,the protein expression of HIF-1α in hypoxia group and DMOG group(480.4±6.91,331±7.78)were significantly upregulated(P＜0.01).Compared with that in DMOG group,the protein expression of HIF-1α in hypoxia group was significantly upregulated(P＜0.01).To compare with the positive cells(51.80±4.15)in control group,the formation of lipid droplets in hypoxia group and DMOG group(27.80±3.11,31.20±2.59)were significantly decreased(P＜0.05).No significant difference was observed on the forma-tion of lipid droplets between that in hypoxia group and that in DMOG group(P＞0.05).Compared with control group,the mRNA expres-sion of PPAR-y and C/EBPα in hypoxia group and DMOG group were significantly decreased(P＜0.01).Compared with hypoxia group,the mRNA expression of PPAR-γ and C/EBPα in DMOG group were significantly decreased(P＜0.05).Compared with control group,the protein expression of PPAR-γ in hypoxia group and DMOG group were decreased 28.3％and 39.8％(P＜0.01).The protein expres-sion of C/EBPα in hypoxia group and DMOG group were decreased 36.7％and 42.5％(P＜0.01).Compared with hypoxia group,the protein expression of PPAR-γ and C/EBPα in DMOG group were decreased 16％and 9.1％(P＜0.05).Conclusion 1％physical hy-poxia and hypoxia mimetic agents DMOG can both inhibit the BMSCs adipogenesis,whereas 1％physical hypoxia had more powerful inhibitory effect than the DMOG.
关键词
低氧/二甲基乙二酰基甘氨酸/成脂肪分化Key words
hypoxia/dimethyloxalylglycine/adipogenesis引用本文复制引用
基金项目
国家自然科学基金(81760392)
云南省科技计划(2018FE001-108)
出版年
2024
NETL
NSTL
万方数据