沉默整合素α6通过激活FOXO1信号通路促进人牙髓干细胞的成牙本质向分化  

Silencing ITGA6 Promotes Odontogenic Differentiation of hDPSCs Through Activating FOXO1 Signaling Pathway

作　　者：王凯丽[1] 张巍巍[1] 李艳萍 何丽娜[1] 张爽 潘爽 牛玉梅[1] WANG Kaili;ZHANG Weiwei;LI Yanping;HE Lina;ZHANG Shuang;PAN Shuang;NIU Yumei(Department of Endodontics,The First Affiliated Hospital of Harbin Medical University,School of Stomatology,Harbin Medical University,Harbin 150001,China)

机构地区：[1]哈尔滨医科大学附属第一医院,哈尔滨医科大学口腔医学院牙体牙髓病科,黑龙江哈尔滨150001

出　　处：《口腔医学研究》2022年第10期930-935,共6页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:81970924)。

摘　　要：目的:探讨沉默整合素α6(integrinα6,ITGA6)是否通过激活FOXO信号通路影响人牙髓干细胞(human dental pulp stem cells,hDPSCs)的成牙本质向分化。方法:利用课题组前期构建的ITGA6沉默慢病毒干扰hDPSCs,通过Real-time PCR和Western blot验证干扰效率,同时检测空载组(sh-NC)和ITGA6沉默组(sh-ITGA6)的FOXO信号通路活性。然后用AS1842856抑制FOXO1,经矿化诱导7 d后进行Western blot和碱性磷酸酶(alkaline phosphatase,ALP)染色来观察hDPSCs的成牙本质向分化情况。结果:Real-time PCR和Western blot结果显示构建的ITGA6沉默慢病毒能有效干扰hDPSCs中ITGA6的表达;与sh-NC组比,sh-ITGA6组FOXO1在mRNA水平表达上调(P<0.05),同时Western blot结果显示其在胞核表达量增加(P<0.05);Western blot检测成牙本质向分化标记物显示,sh-ITGA6组牙本质涎磷蛋白(dentin sialophosphoprotein,DSPP)和牙本质基质蛋白-1(dentin matrix protein-1,DMP-1)表达上调(P<0.05),经AS1842856处理sh-ITGA6后,DSPP和DMP-1表达下降(P<0.05);ALP染色结果显示,sh-ITGA6组ALP染色明显加深,经AS1842856处理sh-ITGA6后,ALP染色变浅。结论:沉默ITGA6通过激活FOXO1信号通路促进hDPSCs的成牙本质向分化。Objective:To investigate whether silencing integrinα6(ITGA6)affects odontogenic differentiation of human dental pulp stem cells(hDPSCs)through activating FOXO signaling pathway.Methods:hDPSCs were transfected with ITGA6 silencing lentiviruses constructed by our research group,real-time PCR and Western blot were applied to evaluate the interference efficiency of ITGA6 silencing lentiviruses.Meanwhile,FOXO signaling pathway activity was detected in empty vector group(sh-NC group)and ITGA6 silencing group(sh-ITGA6 group).FOXO1 was inhibited by AS1842856.Western blot and alkaline phosphatase(ALP)staining were performed after 7 days of mineralization induction to observe the odontogenic differentiation of hDPSCs.Results:ITGA6 silencing lentiviruses effectively inhibited the expression of ITGA6 in hDPSCs.Compared with sh-NC group,the mRNA expression of FOXO1 was upregulated(P<0.05)and Western blot results showed that the expression of FOXO1 in nuclear was increased(P<0.05)in sh-ITGA6 group.The odontogenic differentiation markers DSPP(dental sialophosphoprotein)and DMP-1(dental matrix protein-1)were upregulated in sh-ITGA6 group assessed by Western blot,while the expression of DSPP and DMP-1 were decreased after sh-ITGA6 treated by AS1842856(P<0.05).ALP staining results showed that the ALP staining in sh-ITGA6 group was significantly deeper,however,after treated sh-ITGA6 with AS1842856,ALP staining became shallower.Conclusion:Silencing ITGA6 promoted odontogenic differentiation of hDPSCs through activating FOXO1 signaling pathway.

关 键 词：整合素Α6 牙髓干细胞 FOXO1信号通路 成牙本质向分化 

分 类 号：R780.2[医药卫生—口腔医学]