沉默PGC⁃1α表达对人根尖牙乳头干细胞定向分化的影响  

Effect of silencing PGC⁃1αon the directional differentiation of human stem cells from apical papilla

作　　者：肖雅 于瑞阳 陈路遥 顾婷杰 方玉心 李泽汉 于金华[1] XIAO Ya;YU Ruiyang;CHEN Luyao;GU Tingjie;FANG Yuxin;LI Zehan;YU Jinhua(Department of Endodontics,The Af-filiated Stomatological Hospital of Nanjing Medical University,Jiangsu Province Key Laboratory of Oral Diseases,Jiangsu Province Engi-neering Research Center of Stomatological Translational Medicine,Nanjing 210029,China;School of Pediatrics,Nanjing Medical U-niversity,Nanjing 210029,China)

机构地区：[1]南京医科大学附属口腔医院牙体牙髓科,江苏省口腔疾病研究重点实验室,江苏省口腔转化医学工程研究中心,江苏南京210029 [2]南京医科大学儿科学院,江苏南京210029

出　　处：《口腔生物医学》2023年第3期146-150,共5页Oral Biomedicine

基　　金：国家自然科学基金(82170940);江苏省研究生实践创新计划(JX10513957);江苏省科教能力提升工程——江苏省研究型医院(YJXYYJSDW4);江苏省医学创新中心(CXZX202227)。

摘　　要：目的:探讨过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC⁃1α)在人根尖牙乳头干细胞(hSCAPs)定向分化中的作用。方法:hSCAPs矿化诱导0、3、7 d,实时荧光定量PCR检测PGC⁃1α的基因表达水平。转染小干扰RNA(siRNA)构建稳定低表达PGC⁃1α的hSCAPs,通过实时荧光定量PCR和Western blot检测PGC⁃1α的表达水平,采用碱性磷酸酶(ALP)和茜素红染色分别检测ALP活性和矿化水平的变化,Western blot和实时荧光定量PCR检测成牙/骨向分化相关蛋白和基因的变化。结果:PGC⁃1α的基因表达水平在hSCAPs定向分化过程中逐渐上调。与对照组相比,低表达PGC⁃1α的hSCAPs其ALP表达量下降,矿化结节减少,牙本质涎磷蛋白(DSPP)、ALP、Runt相关转录因子2(RUNX2)、成骨细胞特异性转录因子Osterix(OSX)等成牙/骨向分化相关指标表达水平下调(P<0.05)。结论:沉默PGC⁃1α表达可抑制hSCAPs定向分化。Objective:To explore the effects of silencing peroxisome proliferator⁃activated receptor⁃γcoactivator⁃1α(PGC⁃1α)on the directional differentiation of human stem cells from apical papilla(hSCAPs).Methods:Expression level of PGC⁃1αwas detected by qRT⁃PCR after hSCAPs were cultured in mineralized induction solution for 0,3 and 7 days.hSCAPs with low expression of PGC⁃1αwere constructed stably by siRNA transfection,and the knockdown efficiency of PGC⁃1αwas verified by qRT⁃PCR and Western blot.Further,the effects of silencing PGC⁃1αon the directional differentiation of hSCAPs were observed by alkaline phosphatase(ALP)and alizarin red staining,qRT⁃PCR and Western blot.Results:The expression level of PGC⁃1αwas gradually up⁃regulated during the di⁃rectional differentiation of hSCAPs.As compared with the control group,the mineralized nodules and expression level of ALP in hSCAPs silencing PGC⁃1αwere decreased,and the expression level of indicators related with osteo/odontogenic differentiation(DSPP,ALP,RUNX2,OSX)were downregulated(P<0.05).Conclusions:Silencing PGC⁃1αcan inhibit the directional differentiation ca⁃pacity of hSCAPs.

关 键 词：PGC⁃1α 根尖牙乳头干细胞 RNA干扰 成牙/骨向分化 

分 类 号：R780.2[医药卫生—口腔医学]