PMID- 38241841 OWN - NLM STAT- MEDLINE DCOM- 20240213 LR - 20240213 IS - 1878-1705 (Electronic) IS - 1567-5769 (Linking) VI - 128 DP - 2024 Feb 15 TI - MicroRNA-18a regulates the Pyroptosis, Apoptosis, and Necroptosis (PANoptosis) of osteoblasts induced by tumor necrosis factor-alpha via hypoxia-inducible factor-1alpha. PG - 111453 LID - S1567-5769(23)01780-0 [pii] LID - 10.1016/j.intimp.2023.111453 [doi] AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) is involved in inflammatory responses and promotes cell death and the inhibition of osteogenic differentiation. MicroRNA (miRNA) plays a crucial role in the infected bone diseases, however, the biological role of miRNAs in inflammation-induced impaired osteogenic differentiation remains unclear. This study aimed to explore the role of miRNA-18a-5p (miR-18a) in regulating PANoptosis and osteogenic differentiation in an inflammatory environment via hypoxia-inducible factor-1alpha (HIF1-alpha). METHODS: The expression of miR-18a in MC3T3-E1 cells was analyzed using quantitative reverse transcription-polymerase chain reaction in an inflammatory environment induced by TNF-alpha. The expression of HIF1-alpha and NLRP3 in LV-miR-18a or sh-miR-18a cells was analyzed using western blotting. Fluorescence imaging for cell death, flow cytometry, and alkaline phosphatase activity analysis were used to analyze the role of miR-18a in TNF-alpha-induced PANoptosis and the inhibition of osteogenic differentiation. An animal model of infectious bone defect was established to validate the regulatory role of miR-18a in an inflammatory environment. RESULTS: The expression of miRNA-18a in the MC3T3-E1 cell line was significantly lower under TNF-alpha stimulation than in the normal environment. miR-18a significantly inhibited the expression of HIF1-alpha and NLRP3, and inhibition of HIF1-alpha expression further inhibited NLRP3 expression. Furthermore, inhibition of miR-18a expression promoted the TNF-alpha-induced PANoptosis and inhibition of osteogenic differentiation, whereas miR-18a overexpression and the inhibition of both HIF1-alpha and NLRP3 reduced the effects of TNF-alpha. These findings are consistent with those of the animal experiments. CONCLUSION: miRNA-18a negatively affects HIF1-alpha/NLRP3 expression, inhibits inflammation-induced PANoptosis, and impairs osteogenic differentiation. Thus, it is a potential therapeutic candidate for developing anti-inflammatory strategies for infected bone diseases. CI - Copyright (c) 2023 The Author(s). Published by Elsevier B.V. All rights reserved. FAU - Zhang, Wei AU - Zhang W AD - Department of Orthopaedics, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong, China; The Second School of Clinical Medicine, Southern Medical University. Electronic address: zhangweinlfd@163.com. FAU - Xia, Chang-Liang AU - Xia CL AD - Department of Orthopaedics, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong, China. Electronic address: 986199774@qq.com. FAU - Qu, Yu-Dun AU - Qu YD AD - Department of Orthopaedics, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong, China; The Second School of Clinical Medicine, Southern Medical University. Electronic address: Yudun2685923488@163.com. FAU - Li, Jia-Xuan AU - Li JX AD - Department of Orthopaedics, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong, China. Electronic address: Antares_jiaxuan@163.com. FAU - Liu, Jia-Bao AU - Liu JB AD - Department of Orthopaedics, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong, China. Electronic address: 1492548853@qq.com. FAU - Ou, Shuan-Ji AU - Ou SJ AD - Department of Orthopaedics, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong, China. Electronic address: oushuanji@163.com. FAU - Yang, Yang AU - Yang Y AD - Department of Orthopaedics, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong, China. Electronic address: erjunda@163.com. FAU - Qi, Yong AU - Qi Y AD - Department of Orthopaedics, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong, China; The Second School of Clinical Medicine, Southern Medical University. Electronic address: gd2hqy@163.com. FAU - Xu, Chang-Peng AU - Xu CP AD - Department of Orthopaedics, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong, China; The Second School of Clinical Medicine, Southern Medical University. Electronic address: 731643426@qq.com. LA - eng PT - Journal Article DEP - 20240118 PL - Netherlands TA - Int Immunopharmacol JT - International immunopharmacology JID - 100965259 RN - 0 (Hif1a protein, mouse) RN - 0 (Hypoxia-Inducible Factor 1, alpha Subunit) RN - 0 (MicroRNAs) RN - 0 (NLR Family, Pyrin Domain-Containing 3 Protein) RN - 0 (Tumor Necrosis Factor-alpha) SB - IM MH - Animals MH - Apoptosis MH - *Bone Diseases/metabolism MH - Cell Differentiation MH - Hypoxia-Inducible Factor 1, alpha Subunit/metabolism MH - Inflammation/metabolism MH - *MicroRNAs/genetics MH - Necroptosis MH - NLR Family, Pyrin Domain-Containing 3 Protein/metabolism MH - Osteoblasts/metabolism MH - Osteogenesis MH - Pyroptosis MH - Tumor Necrosis Factor-alpha/metabolism MH - Mice OTO - NOTNLM OT - Hypoxia-inducible factor-1alpha OT - Osteogenic differentiation OT - PANoptosis OT - Tumor necrosis factor-alpha OT - miRNA-18a COIS- Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. EDAT- 2024/01/20 05:42 MHDA- 2024/02/08 06:42 CRDT- 2024/01/19 18:07 PHST- 2023/08/01 00:00 [received] PHST- 2023/12/17 00:00 [revised] PHST- 2023/12/23 00:00 [accepted] PHST- 2024/02/08 06:42 [medline] PHST- 2024/01/20 05:42 [pubmed] PHST- 2024/01/19 18:07 [entrez] AID - S1567-5769(23)01780-0 [pii] AID - 10.1016/j.intimp.2023.111453 [doi] PST - ppublish SO - Int Immunopharmacol. 2024 Feb 15;128:111453. doi: 10.1016/j.intimp.2023.111453. Epub 2024 Jan 18.