碱性成纤维细胞生长因子2对大鼠脂肪干细胞向成牙本质样细胞分化的影响及其机制研究  

Effect of Basic Fibroblast Growth Factor 2 on Differentiation of Rat Adipose Derived Stem Cells into Odontoblast Like Cells and its Mechanism

作　　者：贺会平 姬小婷 HE Hui-ping;JI Xiao-ting(Department of Stomatology,Ninth Hospital of Xi'an,Xi'an Shaanxi 710054)

机构地区：[1]西安市第九医院口腔科,陕西西安710054 [2]陕西中医药大学附属医院口腔科,陕西咸阳712000

出　　处：《医学临床研究》2022年第6期827-830,834,共5页Journal of Clinical Research

基　　金：陕西省提升公众科学素质计划项目(2021PSL135)。

摘　　要：【目的】探讨碱性成纤维细胞生长因子2(FGF-2)对大鼠脂肪干细胞(ADSCs)向成牙本质样细胞分化的影响及其机制。【方法】取大鼠双侧腹股沟处脂肪组织,分离、鉴定ADSCs,取第3代ADSCs随机分为空白对照组(NG组)、阴性对照组(siRNA-NC组)、沉默FGF-2表达组(TGF-2-siRNA组)、通路激活剂组。实时荧光定量PCR法检测各组ADSCs中FGF-2表达水平;CCK-8法检测各组ADSCs增殖情况;采用茜素红染色观察各组ADSCs成牙本质诱导分化情况;采用蛋白免疫印迹法、酶联免疫吸附法(ELISA)法检测神经菌毛素1(NRP1)、牙本质涎蛋白(DSPP)、牙本质基质蛋白1(DMP1)及无翅型MMTV整合位点家族成员蛋白/β-连环素(Wnt/β-catenin)通路蛋白表达水平。【结果】成功分离并鉴定ADSCs;与NG组、siRNA-NC组比较,FGF-2-siRNA组、通路激活剂组ADSCs中FGF-2mRNA、NRP1、DSPP、DMP1、β-catenin蛋白及Wnt水平均降低(P<0.05),ADSCs增殖抑制率升高(P<0.05);与FGF-2-siRNA组比较,通路激活剂组ADSCs中FGF-2mRNA、NRP1、DSPP、DMP1、β-catenin蛋白及Wnt水平均升高(P<0.05),ADSCs增殖抑制率降低(P<0.05)。【结论】FGF-2可影响Wnt/β-catenin通路活化,可能与大鼠ADSCs增殖及向成牙本质样细胞分化有关,可为临床治疗牙齿缺损提供新的思路。【Objective】Objective to investigate the effect of basic fibroblast growth factor-2(FGF-2)on the differentiation of rat adipose derived stem cells(ADSCs)into odontoblast like cells and its mechanism.【Methods】ADSCs were isolated and identified from bilateral inguinal adipose tissue of rats.The third generation ADSCs were randomly divided into blank control group(NG group),negative control group(siRNA-NC group),FGF-2 silencing group(TGF-2-siRNA group)and pathway activator group.The expression level of FGF-2 in ADSCs was detected by real-time fluorescent quantitative PCR;The proliferation of ADSCs in each group was detected by CCK-8 method;Alizarin red staining was used to observe the odontogenic differentiation of ADSCs in each group;Western blotting and enzyme-linked immunosorbent assay(ELISA)were used to detect neuropilin 1(NRP1),dentin sialoprotein(DSPP),dentin matrix protein 1(DMP1)and wingless MMTV integration site family members/β-catenin(Wnt/β-catenin)pathway protein level.【Results】ADSCs were successfully isolated and identified;Compared with the NG group and the siRNA NC group;the expression levels of FGF-2 mRNA,NRP1,DSPP,DMP1,Wnt andβ-catenin protein of the ADSCs in the FGF-2-siRNA group and the pathway activator group decreased(P<0.05),the proliferation inhibition rate of ADSCs increased(P<0.05);Compared with the FGF-2-siRNA group,the expression of FGF-2 mRNA,NRP1,DSPP,DMP1,β-catenin protein and Wnt increased(P<0.05);the proliferation inhibition rate of ADSCs decreased(P<0.05).【Conclusion】FGF-2 can affect Wnt/β-catenin activation pathway.This may be related to the proliferation of rat ADSCs and their differentiation into odontoblast like cells,which may provide a new idea for the treatment of tooth defects.

关 键 词：成纤维细胞生长因子2 干细胞 脂细胞 大鼠 成牙质细胞 

分 类 号：R587[医药卫生—内分泌]