PMID- 37886757 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20231028 IS - 2405-8440 (Print) IS - 2405-8440 (Electronic) IS - 2405-8440 (Linking) VI - 9 IP - 10 DP - 2023 Oct TI - AGEs induce high expression of Dll4 via endoplasmic reticulum stress PERK signaling-mediated internal ribosomal entry site mechanism in macrophages. PG - e21170 LID - 10.1016/j.heliyon.2023.e21170 [doi] LID - e21170 AB - BACKGROUND AND AIM: Advanced glycation end products (AGEs)- exposed macrophages was characterized by Delta-like ligand 4 (Dll4) high expressed and has been shown to participate in diabetes-related atherosclerosis. This study was aimed to investigate the translational regulatory mechanism of Dll4 high expression in macrophages exposed to AGEs. METHODS: Human Dll4 5' untranslated region (5'UTR) sequence was cloned and inserted into a bicistronic reporter plasmid. Human THP-1 macrophages transfected with the bicistronic reporter plasmids were exposed to AGEs. Dual-luciferase assay was used to detect internal ribosome entry site (IRES) activity contained in Dll4 5'UTR. Small interference RNA transfection was used to knock-down specific gene expression. Localization of protein was analyzed. RESULTS: AGEs exposure significantly induced IRES activity in Dll4 5' UTR in human macrophages. Internal potential promoter and ribosome read-through mechanisms were excluded. Inhibition of endoplasmic reticulum stress and specific silencing of protein kinase R-like endoplasmic reticulum kinase (PERK)/eukaryotic initiation factor 2alpha (eIF2alpha) signaling pathway activation reduced IRES activity in Dll4 5' UTR in human macrophages. Dll4 5' UTR IRES activity was also inhibited by targeted silencing of heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1). Moreover, specific inhibition of PERK/eIF2alpha signaling pathway led to deactivation of hnRNPA1, resulting to reduction of AGEs- induced Dll4 5' UTR IRES activity in human macrophages. CONCLUSIONS: AGEs induced Dll4 5' UTR IRES activity in human macrophages which was dependent on endoplasmic reticulum stress PERK/eIF2alpha signaling pathway. hnRNPA1 acted the role as an ITAF was also indispensable for AGEs-induced Dll4 5'UTR IRES activity in human macrophages. CI - (c) 2023 The Authors. FAU - Ma, Yanpeng AU - Ma Y AD - Department of Cardiology, Shaanxi Provincial People's Hospital, Xi'an, 710068, China. AD - Atherosclerosis Integrated Chinese and Western Medicine Key Research Laboratory, Research Office of Shaanxi Administration of Traditional Chinese Medicine, Xi'an, 710003, China. AD - Affiliated Shaanxi Provincial People's Hospital, Northwestern Polytechnical University, Xi'an, 710068, China. FAU - Zheng, Shixiang AU - Zheng S AD - Department of Critical Medicine, Fujian Medical University Union Hospital, Fuzhou, 350000, China. FAU - Wang, Xiqiang AU - Wang X AD - Department of Cardiology, Shaanxi Provincial People's Hospital, Xi'an, 710068, China. AD - Atherosclerosis Integrated Chinese and Western Medicine Key Research Laboratory, Research Office of Shaanxi Administration of Traditional Chinese Medicine, Xi'an, 710003, China. AD - Affiliated Shaanxi Provincial People's Hospital, Northwestern Polytechnical University, Xi'an, 710068, China. FAU - Zhu, Ling AU - Zhu L AD - Department of Cardiology, Shaanxi Provincial People's Hospital, Xi'an, 710068, China. AD - Atherosclerosis Integrated Chinese and Western Medicine Key Research Laboratory, Research Office of Shaanxi Administration of Traditional Chinese Medicine, Xi'an, 710003, China. AD - Affiliated Shaanxi Provincial People's Hospital, Northwestern Polytechnical University, Xi'an, 710068, China. FAU - Wang, Junkui AU - Wang J AD - Department of Cardiology, Shaanxi Provincial People's Hospital, Xi'an, 710068, China. AD - Atherosclerosis Integrated Chinese and Western Medicine Key Research Laboratory, Research Office of Shaanxi Administration of Traditional Chinese Medicine, Xi'an, 710003, China. AD - Affiliated Shaanxi Provincial People's Hospital, Northwestern Polytechnical University, Xi'an, 710068, China. FAU - Pan, Shuo AU - Pan S AD - Department of Cardiology, Shaanxi Provincial People's Hospital, Xi'an, 710068, China. AD - Atherosclerosis Integrated Chinese and Western Medicine Key Research Laboratory, Research Office of Shaanxi Administration of Traditional Chinese Medicine, Xi'an, 710003, China. AD - Affiliated Shaanxi Provincial People's Hospital, Northwestern Polytechnical University, Xi'an, 710068, China. FAU - Zhang, Yong AU - Zhang Y AD - Department of Cardiology, Shaanxi Provincial People's Hospital, Xi'an, 710068, China. AD - Atherosclerosis Integrated Chinese and Western Medicine Key Research Laboratory, Research Office of Shaanxi Administration of Traditional Chinese Medicine, Xi'an, 710003, China. AD - Affiliated Shaanxi Provincial People's Hospital, Northwestern Polytechnical University, Xi'an, 710068, China. FAU - Liu, Zhongwei AU - Liu Z AD - Department of Cardiology, Shaanxi Provincial People's Hospital, Xi'an, 710068, China. AD - Atherosclerosis Integrated Chinese and Western Medicine Key Research Laboratory, Research Office of Shaanxi Administration of Traditional Chinese Medicine, Xi'an, 710003, China. AD - Affiliated Shaanxi Provincial People's Hospital, Northwestern Polytechnical University, Xi'an, 710068, China. LA - eng PT - Journal Article DEP - 20231018 PL - England TA - Heliyon JT - Heliyon JID - 101672560 PMC - PMC10597754 OTO - NOTNLM OT - Advanced glycation end products OT - Delta-like 4 ligand OT - Internal ribosome entry site OT - Macrophages COIS- 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- 2023/10/27 06:42 MHDA- 2023/10/27 06:43 PMCR- 2023/10/18 CRDT- 2023/10/27 04:51 PHST- 2023/05/17 00:00 [received] PHST- 2023/10/14 00:00 [revised] PHST- 2023/10/17 00:00 [accepted] PHST- 2023/10/27 06:43 [medline] PHST- 2023/10/27 06:42 [pubmed] PHST- 2023/10/27 04:51 [entrez] PHST- 2023/10/18 00:00 [pmc-release] AID - S2405-8440(23)08378-0 [pii] AID - e21170 [pii] AID - 10.1016/j.heliyon.2023.e21170 [doi] PST - epublish SO - Heliyon. 2023 Oct 18;9(10):e21170. doi: 10.1016/j.heliyon.2023.e21170. eCollection 2023 Oct.