摘要
目的探究雌激素对人牙周膜成纤维细胞(human periodontal ligament fibroblasts,hPDLF)骨改建的影响及机制。方法取健康正畸患者的前磨牙,提取并鉴定hPDLF细胞;分别用0 nmol/L、0.001 nmol/L、0.01 nmol/L、0.1 nmol/L 17β-雌二醇处理hPDLF细胞24 h后,CCK8法检测细胞增殖能力;qRT-PCR检测骨保护素(OPG)和核因子κB受体活化子配体(RANKL)表达;Western blot检测ERβ表达和ERK1/2磷酸化水平;将ERβ的siRNA转染至hPDLF细胞,然后用0.1 nmol/L17β-雌二醇处理24 h,CCK8法检测细胞增殖能力,qRT-PCR检测OPG和RANKL,Western blot检测ERβ表达和ERK1/2磷酸化水平。结果相比于0 nmol/L雌二醇组,0.001 nmol/L、0.01 nmol/L、0.1 nmol/L17β-雌二醇处理hPDLF细胞后,细胞增殖能力显著增加(P<0.01),OPG表达显著上调(P<0.001),RANKL表达显著下调(P<0.001),ERβ表达和ERK1/2磷酸化水平均显著上调(P<0.001);而转染siRNA抑制ERβ表达后,相比于0.1 nmol/L17β-雌二醇+siRNA组,细胞增殖能力显著下降(P<0.01);OPG表达显著下调(P<0.001),RANKL表达显著上调(P<0.001),ERβ表达和ERK1/2磷酸化水平均显著下调(P<0.001)。结论雌激素通过促进ERβ-ERK1/2信号通路激活而促进人牙周膜成纤维细胞骨改建。
Objective To investigate the effects of estrogen regulation of ERβ-ERK1/2 pathway on reconstruction of human periodontal ligament fibroblast(hPDLF)and its action mechanism.Methods The hPDLF cells were extracted from the premolars of healthy orthodontic patients and identified and then treated with 0 nmol/L,0.001nmol/L,0.01nmol/L and 0.1nmol/L 17β-estradiol,respectively,for 24 hours.CCK-8 assay was used to detect cell proliferation ability.qRT-PCR was used to detect osteoprotectin(OPG)and the expression of nuclear factorκB receptor activator ligand(RANKL).Western blot was used to detect the expression of ERβand the level of ERK1/2 phosphorylation.Moreover,siRNA of ERβwas transfected into hPDLF cells.Then the cells were treated with 0.1nmol/L 17β-estradiol for 24 hours.CCK-8 assay was used to detect cell proliferation ability.qRT-PCR was used to detect osteoprotectin(OPG)and RANKL.Western Blot was used to detect the expression of ERβand the level of ERK1/2 phosphorylation.Results Compared with that in 0nmol/L estradiol group,the cell proliferation ability in the other three groups was significantly increased(P<0.01).And the expression levels of OPG and RANKL were significant up-regulated and down-regulated,respectively(P<0.01).The expression levels of ERβand ERK1/2 phosphorylation were significant up-regulated(P<0.001).After transfection,the cell proliferation ability,compared to that in 0.1nmol/L17β-estradiol+siRNA group,ell proliferation ability was significantly decreased(P<0.01).And the expression levels of OPG were down-regulated significantly(P<0.01),however,the expression levels of RANKL were significantly up-regulated(P<0.01).Moreover the expression levels of ERβand ERK1/2 phosphorylation were down-regulated significantly(P<0.01).Conclusion Estrogen can promote bone reconstruction of human periodontal ligament fibroblasts by improving activation of ERβERK1/2 signaling pathway.
作者
赵玺
李向宇
丁锐
赵浩然
米丛波
吴晓琴
ZHAO Xi;LI Xiangyu;DING Rui(Department of Orthodontics,Stomatological Hospital of Urumqi City,Xinjiang,Urumqi 830000,China)
出处
《河北医药》
CAS
2021年第23期3525-3529,共5页
Hebei Medical Journal
基金
新疆维吾尔自治区自然科学基金(编号:2017D01A18)。
