摘要
目的:探究白血病抑制因子(leukemia inhibitory factor,LIF)对人牙周膜细胞(human periodontal ligament cells,HPDLCs)成骨分化能力的影响。方法:分离培养HPDLCs,分别为空白对照组、标准骨诱导组和加入重组人白血病抑制因子(recombinant human leukemia inhibitory factor,rh LIF)的骨诱导组。7 d后碱性磷酸酶(alkalinephosphatase,ALP)染色及活性检测,21 d后矿化结节染色和定量分析,实时荧光定量PCR(qRT-PCR)检测第5 d HPDLCs中成骨相关基因ALP、骨涎蛋白(bone sialoprotein,BSP)和骨钙蛋白(osteocalcin,OCN)的mRNA表达量。结果:7 d后,标准骨诱导组ALP染色呈深蓝色,LIF刺激后,染色明显变浅,活性显著下降(P<0. 01)。21 d后,标准组HPDLC有大量矿化结节形成,而LIF组明显减少(P<0. 05),qRT-PCR结果显示标准组ALP、BSP和OCN的表达显著提高(P<0. 01),而LIF刺激后,表达明显降低(P<0. 01)。结论:LIF可抑制HPDLC的成骨向分化。
Objective:To probe the effect of leukemia inhibitory factor(LIF)on osteogenic differentiation of human periodontal ligament cells(HPDLCs).Methods:HPDLCs were harvested from orthodontic human and immunohistochemistry staining was employed for morphological observation.Three groups were set in the study including spontaneous differentiation group,bone induction group and LIF supplied bone induction group.After 7 days,alkalinephosphatase(ALP)staining and ALP activity were detected.The expression of ALP,BSP and OCN mRNA was measured by qRT-PCR.After 21 days,mineralized nodules were detected by alizarin red staining.Results:ALP staining appeared a dark blue in the osteogenic induced group,while much weaker in the group treated with LIF.The activity of ALP was significantly higher in the osteogenic induced group than that in the group stimulated by LIF(P<0.01).Alizarin red staining showed lots of mineralized nodules in the osteogenic induced group,while the much less in the group treated with LIF(P<0.05).The mRNA expression of ALP,BSP and OCN was significantly higher in the osteogenic induced group compared to that in the group treated with LIF(P<0.01).Conclusion:LIF inhibited osteoblastic differentiation of HPDLCs in this study.
作者
梁又德
傅润英
刘根
游新
宋大为
Liang You-de;FURun-ying;LIU Gen;YOU Xin;SONG Da-wei(Department of Dentistry,Seventh People’s Hospital of Shenzhen City,Guangdong Shenzhen 518052,China)
出处
《临床口腔医学杂志》
2019年第1期12-15,共4页
Journal of Clinical Stomatology
基金
深圳市盐田区资助项目(20180326)
关键词
白血病抑制因子
牙周膜细胞
成骨
Leukemia inhibitory factor(LIF)
Human periodontal ligament cells(HPDLCs)
Osteogenesis
作者简介
通信作者:梁又德,E-mail:liangyoude1229@sina.com。
