敲低 MALAT1 通过靶向 miR-769-5p/HIF3A 轴抑制慢性牙周炎的进展。

Knockdown of MALAT1 Inhibits the Progression of Chronic Periodontitis via Targeting miR-769-5p/HIF3A Axis.

机构信息

Department of Stomatology, Zibo Central Hospital, No. 54, Gongqingtuan Road, Zhangdian District, Zibo City Shandong Province 255000, China.

出版信息

Biomed Res Int. 2021 Feb 1;2021:8899863. doi: 10.1155/2021/8899863. eCollection 2021.

PURPOSE

Chronic periodontitis (CP) is a long-lasting inflammatory disease that seriously affects oral health. This study is aimed at investigating the regulatory mechanism of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in CP.

METHODS

Primary human periodontal ligament cells (PDLCs) were treated with lipopolysaccharide (LPS) to establish a CP model. Quantitative real-time PCR (qRT-PCR) was used to measure the expression of MALAT1 and miR-769-5p in gingival tissues of patients with CP and LPS-treated PDLCs. Cell viability was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of inflammatory cytokines. The protein levels of caspase-3, Bax, Bcl-2, and hypoxia-inducible factor (HIF) 3A were determined by western blot assay. Dual-luciferase reporter (DLR) assay was applied to validate the target relationships between miR-769-5p and MALAT1/HIF3A.

RESULTS

The expression of MALAT1 and HIF3A was enhanced, and the expression of miR-769-5p was reduced in gingival tissues of patients with CP and LPS-treated PDLCs. MALAT1 knockdown promoted cell viability and inhibited inflammation and cell apoptosis in LPS-treated PDLCs. MALAT1 targeted miR-769-5p and negatively regulated miR-769-5p expression. miR-769-5p overexpression promoted cell viability and inhibited inflammation and cell apoptosis in LPS-treated PDLCs. Besides, miR-769-5p targeted HIF3A and negatively modulated HIF3A expression. Both miR-769-5p inhibition and HIF3A overexpression reversed the inhibitory effects of MALAT1 silencing on LPS-induced PDLC injury in vitro.

CONCLUSION

MALAT1 knockdown attenuated LPS-induced PDLC injury via regulating the miR-769-5p/HIF3A axis, which may supply a new target for CP treatment.

目的

慢性牙周炎（CP）是一种长期存在的炎症性疾病，严重影响口腔健康。本研究旨在探讨转移相关肺腺癌转录物 1（MALAT1）在 CP 中的调控机制。

方法

用脂多糖（LPS）处理原代人牙周膜细胞（PDLCs）建立 CP 模型。采用实时定量 PCR（qRT-PCR）检测 CP 患者牙龈组织和 LPS 处理的 PDLCs 中 MALAT1 和 miR-769-5p 的表达。采用 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐（MTT）法检测细胞活力。酶联免疫吸附试验（ELISA）法检测炎症细胞因子水平。Western blot 法检测 caspase-3、Bax、Bcl-2 和缺氧诱导因子（HIF）3A 的蛋白水平。双荧光素酶报告（DLR）试验验证 miR-769-5p 与 MALAT1/HIF3A 的靶标关系。

结果

CP 患者牙龈组织和 LPS 处理的 PDLCs 中 MALAT1 和 HIF3A 表达增强，miR-769-5p 表达降低。MALAT1 敲低促进 LPS 处理的 PDLC 细胞活力，抑制炎症和细胞凋亡。MALAT1 靶向 miR-769-5p 并负调控 miR-769-5p 表达。miR-769-5p 过表达促进 LPS 处理的 PDLC 细胞活力，抑制炎症和细胞凋亡。此外，miR-769-5p 靶向 HIF3A 并负调控 HIF3A 表达。miR-769-5p 抑制和 HIF3A 过表达逆转了 MALAT1 沉默对 LPS 诱导的 PDLC 损伤的抑制作用。