骨髓间充质干细胞外泌体调控STAT3通路促进牙周膜干细胞成骨分化

口腔生物医学 ›› 2022, Vol. 13 ›› Issue (1): 35-39.

骨髓间充质干细胞外泌体调控STAT3通路促进牙周膜干细胞成骨分化

徐秋¹,邱新毓¹,仇珺¹,高洁²,张浩²

1. 军事口腔医学国家重点实验室国家口腔疾病临床医学研究中心陕西省口腔医学重点实验室
2. 军事口腔医学国家重点实验室国家口腔疾病临床医学研究中心陕西省口腔医学重点实验室第四军医大学口腔医学院口腔正畸学教研室

收稿日期:2021-12-13 修回日期:2022-03-08 出版日期:2022-03-25 发布日期:2022-03-21
通讯作者: 张浩 E-mail:120904448@qq.com
基金资助:
国家自然科学青年科学基金项目;陕西省自然科学基金一般项目-面上项目

Bone marrow mesenchymal stem cell exosomes promote osteogenic differentiation of periodontal ligament stem cells via regulating STAT3 pathway

Received:2021-12-13 Revised:2022-03-08 Online:2022-03-25 Published:2022-03-21
Contact: hao Zhang HaoZhang E-mail:120904448@qq.com

摘要/Abstract

摘要： 目的：研究骨髓间充质干细胞（BMMSCs）外泌体调控信号转导与转录激活因子3（STAT3）通路促进牙周膜干细胞（PDLSCs）成骨分化的作用及机制。方法：原代培养BMMSCs和PDLSCs，取第3代BMMSCs细胞分离外泌体，取第3代PDLSCs细胞进行成骨诱导并分组，外泌体组加入15 μg/mL重悬于磷酸盐缓冲液（PBS）的外泌体，PBS组加入等体积PBS，拮抗剂组加入15 μg/mL外泌体和50 μmol/L以二甲基亚砜（DMSO）为溶剂的STAT3拮抗剂AG490，外泌体+DMSO组加入15 μg/mL外泌体和等体积DMSO，DMSO组加入等体积PBS及DMSO。诱导7 d后，Western blot检测碱性磷酸酶（ALP）、骨钙素（OCN）、Ⅰ型胶原（Col-Ⅰ）、磷酸化酪氨酸激酶2（p-JAK2）、磷酸化STAT3（p-STAT3）的表达；诱导21 d后进行茜素红染色，检测光密度（OD）值。结果：PDLSCs中ALP、OCN、Col-Ⅰ、p-JAK2及p-STAT3的表达和茜素红染色OD值，外泌体组高于PBS组，拮抗剂组低于外泌体+DMSO组（P<0.05）。结论：BMMSCs外泌体通过激活JAK2/STAT3通路促进PDLSCs成骨分化。

Abstract: Objective:To study the effect and mechanism of bone marrow mesenchymal stem cells (BMMSCs) exosomes on promoting osteogenic differentiation of periodontal ligament stem cells (PDLSCs) via regulating signal transducer and activator of transcription 3 (STAT3) pathway. Methods:BMMSCs and PDLSCs were primarily cultured, and the exosomes were isolated from the third generation BMMSCs. The third generation PDLSCs were taken for osteogenic induction and grouped, the exosomes group was treated with 15 μg/mL exosomes and phosphate buffer (PBS), the PBS group was treated with equal volume of PBS, and the antagonist group was treated with 15 μg/mL exosomes and 50 μmol/L STAT3 antagonist AG490 in dimethyl sulfoxide (DMSO) as solvent, (exosome＋DMSO) group was treated with 15 μg/mL exosomes and equal volume of DMSO, DMSO group was treated with equal volume of DMSO and phosphate buffer. The level of A540 was detected. The expression levels of alkaline phosphatase (ALP), osteocalcin (OCN), type I collagen (Col-Ⅰ), phosphorylated janus kinase 2 (p-JAK2) and phosphorylated-STAT3 (p-STAT3) were detected 7 days after induction. Alizarin red staining was performed 21 days after induction, and optical density (OD) was detected. Results:The OD level and the expression levels of ALP, OCN, Col-Ⅰ, p-JAK2 and p-STAT3 of PDLSCs in exosome group were higher than those in PBS group (P<0.05). The results of antagonist group were lower than those in (exosomes＋DMSO) group (P<0.05). Conclusions:BMMSCs exosomes promote the osteogenic differentiation of PDLSCs via activating JAK2/STAT3 pathway.

徐秋邱新毓仇珺高洁张浩. 骨髓间充质干细胞外泌体调控STAT3通路促进牙周膜干细胞成骨分化[J]. 口腔生物医学, 2022, 13(1): 35-39.

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