目的:本研究旨在探讨miR-146a-5p在低能量红色LED照射的hPDLSCs成骨过程中的作用。
方法:用5J/cm2红色LED照射后,实时定量聚合酶链反应(RT-qPCR)检测miR-146a-5p表达,通过RT-qPCR和Western印迹测定成骨标志物的表达。碱性磷酸酶(ALP)活性通过ALP染色评估,通过茜素红染色评估矿化,分别。设计慢病毒载体以调节miR-146a-5p表达。进行双荧光素酶报告基因测定以确认miR-146a-5p和MAPK1之间的靶向关系。短发夹RNA(shRNA)用于调节MAPK1表达。
结果:RT-qPCR和蛋白质印迹显示,5J/cm2的照射提高了hPDLSCs中成骨标志物osterix(OSX)和骨唾液蛋白(BSP)的水平。miR-146a-5p在低能量红色LED光照射下在hPDLSCs中下调。miR-146a-5p低表达显著促进hPDLSCs的成骨潜能。miR-146a-5p靶向MAPK1。5J/cm2的红色LED照射挽救了上调的miR-146a-5p对成骨分化的抑制作用,下调MAPK1可以逆转红色LED照射的积极影响。
结论: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潜在临床辅助疗法。
OBJECTIVE: The study aimed to investigate the role of miR-146a-5p in osteogenesis of hPDLSCs irradiated with low-energy red LEDs.
METHODS: After irradiation with 5 J/cm2 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.
RESULTS: RT-qPCR and western blotting revealed that 5 J/cm2 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/cm2 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.
CONCLUSIONS: 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.