Southwest Medical University, Luzhou, China.
The Department of Preventive Dentistry, The Affiliated Stomatological Hospital, Southwest Medical University, Luzhou, China.
J Periodontal Res. 2024 Oct;59(5):1031-1041. doi: 10.1111/jre.13276. Epub 2024 Jun 6.
The study aimed to investigate the role of miR-146a-5p in osteogenesis of hPDLSCs irradiated with low-energy red LEDs.
After irradiation with 5 J/cm red LED, miR-146a-5p expression was detected by real-time quantitative polymerase chain reaction (RT-qPCR), and osteogenic markers expression was determined by RT-qPCR and Western blotting. Alkaline phosphatase (ALP) activity was assessed by ALP staining, and mineralization was assessed by Alizarin Red staining, respectively. Lentiviral vectors were designed to regulate miR-146a-5p expression. Dual-luciferase reporter assay was performed to confirm the targeted relationship between miR-146a-5p and MAPK1. Short hairpin RNA (shRNA) was used to regulate MAPK1 expression.
RT-qPCR and western blotting revealed that 5 J/cm irradiation elevated the levels of the osteogenic markers osterix (OSX) and bone sialoprotein (BSP) in hPDLSCs. miR-146a-5p is downregulated in hPDLSCs under the low-energy red LED light irradiation. miR-146a-5p underexpression markedly promoted the osteogenic potential of hPDLSCs. miR-146a-5p targeted MAPK1. 5 J/cm red LED irradiation rescued the inhibitory effects of upregulated miR-146a-5p on osteogenic differentiation, and the positive influence of red LED irradiation could be reversed by downregulated MAPK1.
These findings confirm that miR-146a-5p is involved in the effect of LED irradiation on the osteogenic differentiation of hPDLSCs by targeting MAPK1. Red LED irradiation may be a potential clinical adjunct therapy for periodontal regeneration.
本研究旨在探讨 miR-146a-5p 在低能量红光 LED 照射下人牙周膜干细胞成骨中的作用。
用 5J/cm 红光 LED 照射后,通过实时定量聚合酶链反应(RT-qPCR)检测 miR-146a-5p 的表达,通过 RT-qPCR 和 Western blot 检测成骨标志物的表达。通过碱性磷酸酶(ALP)染色评估 ALP 活性,通过茜素红染色分别评估矿化。设计慢病毒载体来调节 miR-146a-5p 的表达。通过双荧光素酶报告基因实验证实 miR-146a-5p 与 MAPK1 之间的靶向关系。使用短发夹 RNA(shRNA)调节 MAPK1 的表达。
RT-qPCR 和 Western blot 显示,5J/cm 照射可提高 hPDLSCs 中成骨标志物osterix(OSX)和骨涎蛋白(BSP)的水平。低能量红光 LED 照射下 hPDLSCs 中 miR-146a-5p 下调。miR-146a-5p 低表达显著促进 hPDLSCs 的成骨潜能。miR-146a-5p 靶向 MAPK1。5J/cm 红光 LED 照射可挽救上调 miR-146a-5p 对成骨分化的抑制作用,下调 MAPK1 可逆转红光 LED 照射的正向影响。
这些发现证实,miR-146a-5p 通过靶向 MAPK1 参与 LED 照射对 hPDLSCs 成骨分化的影响。红光 LED 照射可能是牙周再生的一种潜在临床辅助治疗方法。