Dongfeng Stomatological Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, China.
Comput Math Methods Med. 2022 Apr 25;2022:4634925. doi: 10.1155/2022/4634925. eCollection 2022.
miR-141-3p has been demonstrated to be both anti-inflammatory and osteoprotective. This study is aimed at investigating the effect of miR-141-3p on osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) stimulated by Porphyromonas gingivalis lipopolysaccharide (PgLPS) and its mechanism.
PgLPS was used to induce an inflammatory environment, and overexpression of miR-141-3p was done to assess its effect on hPDLSCs in an inflammatory environment. The level of miR-141-3p and EZH2 in hPDLSCs from each treatment group was detected via qRT-PCR, and the inflammatory factors IL-6 and IL-8 in the supernatant of each group were detected by ELISA. ALP staining and alizarin red staining were used to assess the effect of miR-141-3p on the osteogenic differentiation ability of hPDLSCs, and also, western blot was used to detect expression of osteogenic differentiation-related proteins. Further, dual-luciferase reporter assay examined whether miR-141-3p targeted EZH2.
PgLPS led to a significant decrease of miR-141-3p in hPDLSCs. Overexpression of miR-141-3p could enhance ALP activity and alizarin red staining intensity and increase Runx2, OPN and OCN protein expression levels in PgLPS-treated hPDLSCs. Additionally, miR-141-3p could reduce IL-6 and IL-8. miR-141-3p could target and negatively regulate EZH2, and overexpression of EZH2 reversed the promoting effect of miR-141-3p on osteogenic differentiation.
miR-141-3p can attenuate PgLPS-induced inhibition of osteogenic differentiation and inflammation in hPDLSCs by negatively regulating EZH2.
miR-141-3p 具有抗炎和护骨作用。本研究旨在探讨 miR-141-3p 对牙龈卟啉单胞菌脂多糖(PgLPS)刺激下人牙周膜干细胞(hPDLSCs)成骨分化的影响及其机制。
用 PgLPS 诱导炎症环境,过表达 miR-141-3p 以评估其在炎症环境下对 hPDLSCs 的影响。通过 qRT-PCR 检测各组 hPDLSCs 中 miR-141-3p 和 EZH2 的水平,ELISA 检测各组上清液中炎症因子 IL-6 和 IL-8 的水平。ALP 染色和茜素红染色评估 miR-141-3p 对 hPDLSCs 成骨分化能力的影响,Western blot 检测成骨分化相关蛋白的表达。进一步通过双荧光素酶报告实验检测 miR-141-3p 是否靶向 EZH2。
PgLPS 导致 hPDLSCs 中 miR-141-3p 显著降低。过表达 miR-141-3p 可增强 PgLPS 处理的 hPDLSCs 中的 ALP 活性和茜素红染色强度,并增加 Runx2、OPN 和 OCN 蛋白表达水平。此外,miR-141-3p 可降低 IL-6 和 IL-8。miR-141-3p 可靶向并负调控 EZH2,过表达 EZH2 逆转了 miR-141-3p 对成骨分化的促进作用。
miR-141-3p 通过负调控 EZH2 减轻 PgLPS 诱导的 hPDLSCs 成骨分化和炎症抑制。