miR-21对牙周膜干细胞炎症状态及成骨分化的影响

doi:10.3969/j.issn.1674-8603.2020.02.005

摘要/Abstract

摘要： 目的：研究miR-21对牙周膜干细胞（PDLSCs）炎症状态及成骨分化的影响。方法：收集牙周炎患者和健康人牙龈组织，实时荧光定量PCR（qRT-PCR）检测miR-21的表达。采用10?ng/mL肿瘤坏死因子-α（TNF-α）刺激PDLSCs，模拟体外炎症微环境，qRT-PCR检测miR-21的表达。分别采用miR-21模拟物和miR-21抑制剂对miR-21进行过表达和敲低，qRT-PCR检测miR-21对TNF-α诱导的PDLSCs炎性因子IL-6、IL-1β分泌的影响。通过qRT-PCR和碱性磷酸酶（ALP）染色检测miR-21在PDLSCs成骨分化中的调控作用。结果：qRT-PCR结果显示miR-21在牙周炎患者牙龈组织中表达升高（P＜0.05）；TNF-α刺激后PDLSCs中miR-21表达上调（P＜0.01）。过表达miR-21后，PDLSCs中IL-6、IL-1β表达量降低（P＜0.05），敲低miR-21促进IL-6、IL-1β的表达（P＜0.05）。PDLSCs成骨诱导过程中，miR-21表达量显著升高（P＜0.01）。过表达miR-21后，成骨相关基因RUNX2和OCN表达上调，ALP染色增强（P＜0.01）；敲低miR-21后成骨相关基因RUNX2和OCN表达下调，ALP染色减弱（P＜0.05）。在TNF-α刺激下，PDLSCs成骨能力减弱，过表达miR-21后PDLSCs成骨相关基因RUNX2表达上调，ALP染色增强（P＜0.05）。结论：miR-21抑制TNF-α诱导的PDLSCs炎性因子的表达，促进炎症微环境中PDLSCs的成骨分化。

关键词: miR-21, 牙周膜干细胞, 炎症反应, 成骨向分化, miR-21, 牙周膜干细胞, 炎症反应, 成骨向分化

Abstract: Objective：To explore the role of miR-21 in inflammatory response and osteogenic differentiation ofperiodontal ligament stem cell (PDLSCs). Methods：Gingiva samples from periodontitis patients and healthy people were collected and real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed to detect the expression of miR-21. 10 ng/mLtumor necrosis factor (TNF-α) was used to stimulate PDLSCs, mimicking inflammatory environment in vitro. QRT-PCR was performed to detect the expression of miR-21. MiR-21 mimics and inhibitors were utilized respectively to overexpress and knockdown miR-21 in PDLSCs. QRT-PCR and alkaline phosphatase (ALP) staining were performed to confirm the role of miR-21 on the osteogenic differentiation of PDLSCs. Results：The expression of miR-21 was upregulated in the gingiva tissues from periodontitis patients(P＜0.05). The miR-21 expression in PDLSCs was also upregulated under TNF-α stimulation (P＜0.01). Overexpression of miR-21 down-regulated the expression of cytokines IL-6, IL-1βinduced by TNF-α in PDLSCs(P＜0.05). Conversely, knocking down miR-21 promoted the expression of those genes(P＜0.05). During the osteogenesis induction of PDLSCs, the expression of miR-21 was significantly increased (P＜0.01). Overexpression of miR-21 enhanced and knocking down miR-21 alleviated the expression of osteogenic related genes RUNX2, OCNand the ALP staining (P＜0.05). In the inflammatory microenvironment, miR-21 overexpression could alleviate the inhibition effect of TNF-α stimulation on osteogenesis of PDLSCs (P＜0.05).Conclusions：MiR-21 alleviates the expression of cytokines induced by TNF-α in PDLSCs and promotes osteogenic differentiation of PDLSCs in the inflammatory microenvironment.

Key words: miR-21, periodontal ligament stem cells, miR-21, periodontal ligament stem cells, inflammation response, inflammation response, osteogenic differentiation, osteogenic differentiation

杨乔林刘朋郑云飞李巍然. miR-21对牙周膜干细胞炎症状态及成骨分化的影响[J]. 口腔生物医学, 2020, 11(2): 86-91.

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