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作 者:陶庭亮[1] 邓超[2] 柳海[2] 周嵩琳[2] 徐清[2] 王云[2]
机构地区:[1]合肥市口腔医院,安徽合肥230001 [2]皖南医学院口腔学院
出 处:《口腔医学研究》2015年第5期500-502,505,共4页Journal of Oral Science Research
基 金:皖南医学院重点科研项目培育基金(编号:WK2013Z10)
摘 要:目的:探讨糖基化终末产物(AGEs)对人牙周膜干细胞(HPDLSC)增殖能力以及增殖相关基因HSG、cyclinD1的影响。方法:体外组织块法和有限稀释法克隆化培养牙周膜干细胞;成骨、成脂诱导牙周膜细胞,对其进行干细胞鉴定;将培养出的牙周膜干细胞与不同浓度的AGEs共培养,MTT检测不同浓度下牙周膜干细胞增殖的改变;实时定量聚合酶链反应(real time PCR)检测AGEs刺激后HSG、cyclin D1表达的改变。结果:牙周膜细胞成骨诱导21d后茜素红染色出现钙化结节;成脂诱导21d后油红O染色出现脂滴;MTT显示不同浓度的AGEs对HPDLSCs增殖能力的影响有所差别,高浓度(100mg/L,200mg/L)明显抑制HPDLSCs的增殖,差异有统计学意义(P<0.05),低浓度(1mg/L,10mg/L)对HPDLSCs的增殖能力影响不大,差异无统计学意义;Real time PCR结果显示:AGEs(100mg/L)刺激3d后HSG mRNA表达水平较对照组升高、cyclinD1mRNA的表达水平较对照组降低,差异有统计学意义(P<0.05)。结论:高浓度的AGEs能抑制人牙周膜干细胞的增殖并能改变HSG、cyclinD1mRNA的表达。Objective:To investigate the effect of advanced glycation end products(AGEs)on the proliferation of human periodontal ligament stem cells(HPDLSCs).Methods:HPDLSCs were isolated by limited dilution of culture cells for single cell clone.The osteogenic differentiation capacity of HPDLSCs was evaluated by Alizarin red staining.The adipogenic differentiation capacity of HPDLSCs was evaluated by oil red staining.HPDLSCs were induced with different concentrations of AGEs,The proliferation of HPDLSCs was assayed by MTT,Real time quantitative reverse transcription polymerase chain reaction(real time PCR)was performed to detect the differences of gene expression between the control group and experimental group.Results:After 21 days induction,Alizarin red staining showed mineralization nodules were formed,oil red staining showed lipid droplets were formed.Different concentrations of AGEs had different effects on the PDLSCs proliferative capacity.High concentrations(100mg/L,200mg/L)significantly inhibited the proliferation of PDLSCs.Low concentration(1mg/L,10mg/L)had little effect on the proliferative capacity of PDLSCs.After 3days,the expressions of cell cycle gene(cyclinD1)in the experimental group were lower than those in the control group,the expressions of HSG in the experimental group were higher than those in the control group(P〈0.05).Conclusion:High concentrations of AGEs reduced the proliferation capacity of HPDLSCs,and changed the expressions of HSG and cyclinD1 mRNA levels.
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