雌激素受体β对牙周膜细胞骨向分化能力影响的研究  

Effects of estrogen receptor beta on osteoblastic differentiation function of human periodontal ligament cells

作　　者：梁莉[1] 周威[1] 余继锋[2] 温莉莎[1] 丁寅 

机构地区：[1]解放军309医院口腔科,北京100091 [2]第二炮兵总医院眼科,北京100088 [3]第四军医大学口腔医院正畸科,西安710032

出　　处：《口腔医学》2010年第9期513-516,共4页Stomatology

基　　金：国家自然科学基金(30572069)

摘　　要：目的研究小干扰RNA(small interfering RNA,siRNA)抑制人牙周膜成纤维细胞(human periodontal ligament fibroblast cell,HPLF)中雌激素受体β(estrogen receptor β,ERβ)后,对17β-雌二醇(E2)诱导的成骨能力的影响。方法设计并合成针对ERβ基因siRNA的寡核苷酸序列,插入载体形成重组载体。重组载体经测序鉴定后,以脂质体法转染至HPLF细胞中,蛋白质印迹法及RT-PCR法检测转染前后ERβ的表达情况。鉴定后用1×10-7mol/L的17β-E2干预经ERβsiRNA转染的HPLF细胞和未经ERβsiRNA转染的HPLF细胞,测定细胞的碱性磷酸酶(alkaline phosphatase,ALP)活性和骨钙素(os-teocalcin,OCN)含量。结果测序鉴定重组质粒后,蛋白质印迹法及RT-PCR结果显示ERβsiRNA载体可特异地抑制HPLF细胞中ERβ的表达。经雌激素干预后,HPLF细胞ALP活性和OCN含量高于对照组(P<0.01),但ERβsiRNA载体稳定转染的HPLF细胞ALP活性和OCN含量与对照组相比没有显著性差异。结论雌激素可能通过ERβ亚基促进人牙周膜成纤维细胞的骨向分化能力。Objective To investigate the effects of ERβ on osteoblastic differentiation function of HPLF cells by measuring the alkaline phosphatase（ALP） activity and the production of osteocalcin（OCN） in vitro.Methods We employed a short interfering RNA（siRNA） technique to inhibit ERβ expression in HPLF cells.The cells were cultured with a saturating concentration of 17β-estradiol（10-7 mol/L）.ALP activity was analysed by colorimetric assay using ALP kit and the amount of OCN was assessed by osteocalcin ELISA kit.Results The results of the RT-PCR and Western blot analysis showed that ERβ mRNA and protein expression were inhibited and stabilized transfected cells were constructed successfully.A clear increase of ALP activity and the levels of OCN were observed after estradiol incubation in HPLF cells（P0.05）.However,no evident changes of ALP activity and the levels of OCN could be observed in HPLF-siERβ cells after estradiol treatment.Conclusions These results indicate that estrogen may influence the bone formation capacity of HPLF cells mainly via ERβ.

关 键 词：牙周膜细胞 雌激素受体Β 小干扰RNA 碱性磷酸酶 骨钙素 

分 类 号：R347.931[医药卫生—基础医学]