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

doi:10.13701/j.cnki.kqyxyj.2022.10.006

口腔医学研究 ›› 2022, Vol. 38 ›› Issue (10): 930-935.DOI: 10.13701/j.cnki.kqyxyj.2022.10.006

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

王凯丽, 张巍巍, 李艳萍, 何丽娜, 张爽, 潘爽, 牛玉梅^*

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

收稿日期:2022-03-21 出版日期:2022-10-28 发布日期:2022-10-20
通讯作者: *牛玉梅,E-mail:yumeiniu@163.com
作者简介:王凯丽(1994~ ),女,山东人,医师,硕士,主要从事牙髓干细胞再生的相关研究。
基金资助:
国家自然科学基金(编号:81970924)

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

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

Received:2022-03-21 Online:2022-10-28 Published:2022-10-20

摘要/Abstract

摘要： 目的: 探讨沉默整合素α6(integrin α6,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。

关键词: 整合素α6, 牙髓干细胞, FOXO1信号通路, 成牙本质向分化

Abstract: Objective: To investigate whether silencing integrin α6 (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.

Key words: integrin α6, dental pulp stem cells, FOXO1 signaling pathway, odontogenic differentiation

王凯丽, 张巍巍, 李艳萍, 何丽娜, 张爽, 潘爽, 牛玉梅. 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[J]. 口腔医学研究, 2022, 38(10): 930-935.

WANG Kaili, ZHANG Weiwei, LI Yanping, HE Lina, ZHANG Shuang, PAN Shuang, NIU Yumei. Silencing ITGA6 Promotes Odontogenic Differentiation of hDPSCs Through Activating FOXO1 Signaling Pathway[J]. Journal of Oral Science Research, 2022, 38(10): 930-935.

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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

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