雌激素对人牙周膜干细胞干性维持的影响  

Effects of estrogen on the stemness maintenance of human periodontal ligament stem cells

作　　者：戴旭彬 王萧萧[1] 杨凡巧 欧乾民 姚斯琦 王彦[1] 林雪峰[1] 

机构地区：[1]中山大学光华口腔医学院·附属口腔医院,广东省口腔医学重点实验室,广州510055

出　　处：《中华口腔医学研究杂志（电子版）》2016年第2期104-111,共8页Chinese Journal of Stomatological Research(Electronic Edition)

基　　金：国家自然科学基金(81171575)

摘　　要：目的研究雌激素对人牙周膜干细胞(h PDLSC)干性维持的影响。方法分离、培养原代h PDLSC并完成其干细胞鉴定;雌激素处理48 h,采用流式细胞术检测雌激素对h PDLSC细胞周期的影响;定量聚合酶链反应(q-PCR)检测人端粒酶逆转录酶(h TERT)基因、干性相关基因Oct4、Sox2、c-Myc以及衰老相关基因p16和p53的变化;雌激素长期处理后观察细胞克隆形成能力及骨向分化能力的改变。应用SPSS 17.0软件,利用配对样本t检验对组间均数进行统计学分析,P<0.05为差异具有统计学意义。结果实验获得的h PDLSC符合间充质干细胞鉴定标准。流式细胞术结果显示,雌激素处理48 h后h PDLSC增殖能力(29.730±1.845)较对照组(23.587±2.905)明显提高,差异有统计学意义(t=9.913,P=0.010)。q-PCR结果显示雌激素处理48 h后,h TERT表达量(1.958±0.338)较对照组(1.000±0.018)明显提高(t=7.00,P=0.001);Oct4表达量(2.539±0.493)较对照组(1.000±0.011)明显提高(t=6.26,P=0.001);Sox2表达量(2.234±0.255)较对照组(1.016±0.221)明显提高(t=6.26,P=0.003);c-Myc表达量(1.328±0.091)较对照组(1.003±0.088)明显提高(t=5.67,P=0.002);而衰老相关基因p16表达量(0.460±0.085)较对照组(1.009±0.163)明显降低(t=12.24,P=0.007);p53表达量(0.301±0.041)较对照组(1.004±0.115)明显降低(t=7.77,P=0.016)。雌激素长期处理后,雌激素处理组的克隆形成率(0.058±0.008)显著高于对照组(0.013±0.008),差异有统计学意义(t=5.60,P=0.028),成骨能力显著强于对照组(A_(对照组)=1.238±0.084;A_(E2)=2.460±0.182,t=16.41,P<0.001)。结论雌激素能提高h TERT和Oct4、Sox2、c-Myc的基因表达量,维持细胞增殖,抑制细胞衰老。雌激素长期处理能维持体外扩增时h PDLSC的干性。Objective To investigate the biological effect of estrogen（E2） on the stemnessmaintenance of human periodontal ligament stem cells（h PDLSCs）. Methods Primary h PDLSCs wereisolated and characterized. After treatment with estrogen for 48 h,flow cytometry was used to evaluate cellcycle. The changes in human telomerase reverse transcriptase（h TERT）genes,stemness related genesOct4,Sox2,c-Myc,aging related genes p16 and p53 were detected by q-PCR. After long-term culture withestrogen,colony formation ability and osteogenic differentiation potential were evaluated. The mean valuesbetween groups were analyzed by Paired-Samples T Test in SPSS 17.0. A significance level was set at 0.05.Results We isolated h PDLSCs and confirmed their capacity as mesenchymal stem cells. Proliferationability of h PDLSCs was increased under 48 hours estrogen treatment（t = 9.913,P = 0.010）. q-PCR resultsshowed that the expression level of h TERT（1.958 ± 0.338）was higher than the control group（1.000 ±0.018）,there was a statistically significant difference between the groups（t = 7.00,P = 0.001）;theexpression level of Oct4（2.539 ± 0.493）was higher than the control group（1.000 ± 0.011）,there was astatistically significant difference between the groups（t = 6.26,P = 0.001）;the expression level of Sox2（2.234 ± 0.255）was higher than the control group（1.016 ± 0.221）,there was a statistically significant difference between the groups（t = 6.26,P = 0.003）;the expression level of c-Myc（1.328 ± 0.091）was higher than the control group（1.003 ± 0.088）,there was a statistically significant difference between the groups（t = 5.67,P = 0.002）,while senescence related genes that the expression level of p16（0.460 ±0.085）was lower than the control group（1.009 ± 0.163）,there was a statistically significant difference between the groups（t = 12.24,P = 0.007）and the expression level of p53（0.301 ± 0.041）was lower than the control group（1.004 ± 0.115）,there was a statistically s

关 键 词：雌激素 人牙周膜干细胞 干性 人端粒酶逆转录酶 

分 类 号：R781.4[医药卫生—口腔医学]