摘要
目的探讨Notfh信号通路对大鼠牙囊细胞(rDFCs)增殖和成骨分化的调控作用t方法体外培养rDFCs,经细胞形态、免疫组化鉴定rDFCs后,使用lOng/L大鼠重组Jagged1蛋白、10mol/LDAPT溶液、对照组培养液作用于细胞,CCK-8检测增殖能力;利用碱性磷酸酶、茜素红染色分析rDFCs成骨分化能力;Western Biol检测关键蛋白Notch受体胞内段(NICD)、碱性磷酸酶(ALP)、骨形成蛋白2(BMP2)。结果rDFCs呈成纤维细胞形态;rDFCs中波形丝蛋白明显表达,角蛋白未表达;诱导组ALP染色、茜素红染色均强于对照组;Jagg«ll第4天后能明显促进出只^细胞增殖,DAPT第4天后能显著抑制rDFCs(P<0.05)。Jagged1组ALP、BMP2的表达趋势下降,DAPT组ALP、BMP2的表达趋势升高(P<0.05)。结论激活Notch信号通路可促进rDFCs的增殖,抑制Notch信号通路可促进rDFCs的成骨分化。
Objective To investigate the regulatory effect of Notch signal pathway on the proliferation and osteogenic differentiation of rat dental follicle cells(rDFCs). Methods RDFCs were cultured and the cells were identified by cell morphology and immunohistochemistry. The cells were treated with 10 ng/L rat recombinant Jagged1 protein, 10 μmol/LDAPT solution and control group medium.CCK-8 was used to detect cell proliferation. The osteogenic differentiation ability of rDFCs was analyzed by alkaline phosphatase and alizarin red staining, and the key protein Notch receptor(NiCd),alkaline phosphatase(ALP) and bone morphogenetic protein 2(BMP 2) were detected by Western blot. Results rDFC showed fibroblast morphology.The expression of filament protein in rDFC was obvious, but the keratin was not expressed.ALP staining and alizarin red staining in the induction group were stronger than those in the control group.Jagged1 could significantly promote the proliferation of rDFC cells after 4 days,while DAPT could significantly inhibit the proliferation of rDFC cells after 4 days(P<0.05).The expression of ALP and BMP2 decreased in Jagged group,but increased in DAPT group(P<0.05). Conclusions Activation of Notch signaling pathway can promote the proliferation of rDFCcs, and inhibition of Notch signaling pathway can promote the osteogenic differentiation of rDFCs.
作者
赵刚
段序
王丹
丁元圣
ZHAO Gang;DU AN Xu;WANG Dan;DING Yuan-sheng(School of Stomatology,Jiamusi University,Jiamusi154000,China)
出处
《北京口腔医学》
CAS
2021年第5期271-275,共5页
Beijing Journal of Stomatology
基金
黑龙江省卫生健康委科研课题(2019-320)。
关键词
NOTCH信号通路
牙囊细胞
细胞增殖
成骨分化
Notch signaling pathway
Dental follicle cells
Cell proliferation
Osteogenic differentiation
