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miR-141-3p 通过调控 EZH2 减轻牙龈卟啉单胞菌脂多糖诱导的人牙周膜干细胞炎症反应和抑制成骨分化。

miR-141-3p Regulates EZH2 to Attenuate Porphyromonas gingivalis Lipopolysaccharide-Caused Inflammation and Inhibition of Osteogenic Differentiation in Human Periodontal Ligament Stem Cells.

机构信息

Dongfeng Stomatological Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, China.

出版信息

Comput Math Methods Med. 2022 Apr 25;2022:4634925. doi: 10.1155/2022/4634925. eCollection 2022.

Abstract

OBJECTIVE

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.

METHODS

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.

RESULTS

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.

CONCLUSION

miR-141-3p can attenuate PgLPS-induced inhibition of osteogenic differentiation and inflammation in hPDLSCs by negatively regulating EZH2.

摘要

目的

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。

方法

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。

结果

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。

结论

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6cd1/9061008/66a9f0093f47/CMMM2022-4634925.001.jpg

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