山柰酚对炎症状态下人根尖乳头干细胞成骨/成牙本质向分化的影响及相关机制研究  

The effect of kaempferol on the osteogenic/dentinogenic differentiation of stem cells from the human apical papilla under inflammation and its related mechanism

作　　者：赵璐[1] 殷婷 孙江伟 吴佩玲[1] ZHAO Lu;YIN Ting;SUN Jiang-wei;WU Pei-ling(Department of Stomatology,The Second Affiliated Hospital of Xinjiang Medical University,Xinjiang Urumqi 830063,China.)

机构地区：[1]新疆医科大学第二附属医院口腔科,新疆乌鲁木齐830063

出　　处：《临床口腔医学杂志》2024年第12期707-712,共6页Journal of Clinical Stomatology

基　　金：新疆维吾尔自治区自然科学基金项目(编号:2022D01C786)。

摘　　要：目的:研究山柰酚对炎症状态下人根尖乳头干细胞(stem cells from the apical papilla,SCAP)的成骨/成牙本质向分化的影响及Wnt/β-catenin信号通路的调控作用。方法:采用酶消化法结合组织块法获得SCAP,流式细胞术检测细胞表面标志物。成骨诱导2周后,设空白对照组,TNF-α组,TNF-α+XAV939组,TNF-α+山柰酚组,TNF-α+山柰酚组+XAV939组进行ALP的活性水平测定,茜素红染色及荧光定量检测,Western blot分析检测通路相关蛋白表达情况。结果:与TNF-α组相比较,TNF-α+0.1μmol/L山柰酚组ALP活性显著提高,而TNF-α+XAV939组降低(P<0.05)。与TNF-α+0.1μmol/L山柰酚组比较,TNF-α+0.1μmol/L山柰酚+XAV939组ALP活性表达抑制(P<0.05)。未加入山柰酚的TNF-α+XAV939组较TNF-α+0.1μmol/L山柰酚+XAV939组,茜素红染色明显变浅,矿化结节形成的数量小且稀疏。与TNF-α组相比较,TNF-α+0.1μmol/L山柰酚组DSPP、OPN、RUNX2、OCN表达水平均显著提高,而TNF-α+XAV939组均降低(P<0.05);与TNF-α+0.1μmol/L山柰酚组比较,TNF-α+0.1μmol/L山柰酚+XAV939组DSPP、OPN、RUNX2、OCN表达水平均降低(P<0.05)。TNF-α组成骨相关蛋白和通路相关蛋白β-catenin、p-GSK3β的表达水平均高于TNF-α+XAV939组(P<0.05);与TNF-α+0.1μmol/L山柰酚组比较,TNF-α+0.1μmol/L山柰酚+XAV939组β-catenin、p-GSK3β的表达水平均降低(P<0.05);GSK3β表达水平无明显差异。结论:浓度0.1μmol/L山柰酚能促进炎症状态下SCAP成骨/成牙本质向分化的潜能,Wnt/β-catenin信号通路在此过程中发挥重要作用。Objective:To study the effect of kaempferol on the osteogenic/dentinogenic differentiation of stem cells from the human apical papilla(SCAP)under inflammation and the regulation of Wnt/β-catenin signaling pathway.Methods:SCAP was obtained by enzymatic digestion combined with tissue block method,and cell surface markers were detected by flow cytometry.After 2 weeks of osteogenic induction,they were divided into blank control group,TNF-αgroup,TNF-α+XAV939 group,TNF-α+kaempferol group,TNF-α+kaempferol group+XAV939 group.Alkaline phosphatase(ALP)staining activity assay,alizarin red staining and real-time RT-PCR,Western blot analysis to detect the expression of pathway-related proteins.Results:Compared with the TNF-αgroup,ALP activity was significantly increased in the TNF-α+0.1μmol/L kaempferol group,decreased in the TNF-α+XAV939 group(P<0.05).The expression of ALP activity was inhibited in the TNF-α+0.1μmol/L kaempferol+XAV939 group compared with the TNF-α+0.1μmol/L kaempferol group(P<0.05).Alizarin red staining was significantly lighter and the number of mineralized nodules formed was small and sparse in the TNF-α+XAV939 group without added kaempferol compared with the TNF-α+0.1μmol/L kaempferol+XAV939 group.Compared with the TNF-αgroup,the expression levels of DSPP,OPN,RUNX2,and OCN were significantly increased in the TNF-α+0.1μmol/L kaempferol group,while they were all decreased in the TNF-α+XAV939 group(P<0.05).The expression levels of DSPP,OPN,RUNX2,and OCN were all reduced in the TNF-α+0.1μmol/L kaempferol+XAV939 group compared with the TNF-α+0.1μmol/L kaempferol group(P<0.05).The expression levels of osteogenesisrelated proteins and pathway-related proteinsβ-catenin and p-GSK3βwere higher in the TNF-αgroup than in the TNF-α+XAV939 group(P<0.05).Compared with the TNF-α+0.1μmol/L kaempferol group,the expression levels ofβ-catenin and p-GSK3βwere reduced in the TNF-α+0.1μmol/L kaempferol+XAV939 group(P<0.05).And there was no significant difference in GSK3βexpression level among

关 键 词：根尖乳头干细胞 肿瘤坏死因子-Α 山柰酚 WNT/Β-CATENIN 

分 类 号：Q813.1[生物学—生物工程]