Omeprazole exacerbates intervertebral disc degeneration through Caspase-3 mediated apoptosis of nucleus pulposus cells: a Mendelian randomization, network toxicology, and in vitro experimental study

Abstract Objective To investigate the causal correlation and toxicological mechanisms of omeprazole in intervertebral disc degeneration (IVDD), alongside a particular emphasis on Caspase-3 (CASP3) mediated apoptosis of nucleus pulposus cells (NPCs). Methods Mendelian randomization (MR): GWAS data wa...

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Main Authors: Yuchao Jia, Haifan Zhao, Shengbo Huang, Baoshan Xu
Format: Article
Language:English
Published: BMC 2025-05-01
Series:Journal of Orthopaedic Surgery and Research
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Online Access:https://doi.org/10.1186/s13018-025-05863-4
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author Yuchao Jia
Haifan Zhao
Shengbo Huang
Baoshan Xu
author_facet Yuchao Jia
Haifan Zhao
Shengbo Huang
Baoshan Xu
author_sort Yuchao Jia
collection DOAJ
description Abstract Objective To investigate the causal correlation and toxicological mechanisms of omeprazole in intervertebral disc degeneration (IVDD), alongside a particular emphasis on Caspase-3 (CASP3) mediated apoptosis of nucleus pulposus cells (NPCs). Methods Mendelian randomization (MR): GWAS data was employed to assess causal associations between proton pump inhibitors (PPIs) and IVDD. Network toxicology: Shared omeprazole-IVDD targets were identified using STRING, SwissTargetPrediction, and GeneCards databases. Functional enrichment analysis: Biological pathways were explored by employing Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Molecular docking: Omeprazole-CASP3 binding affinity was assessed by employing AutoDock Vina. Experimental validation: Rat NPCs were subjected to CCK-8 assay viability, flow cytometry apoptosis, Western blot, and immunofluorescence. Results MR analysis suggested omeprazole substantially augmented IVDD risk (OR = 1.058, 95% CI = 1.004–1.115, P = 0.034), with no association observed for esomeprazole or lansoprazole. Network toxicology identified 11 overlapping targets, with CASP3 as the hub gene. Molecular docking revealed strong omeprazole-CASP3 binding (free energy: − 6.725 kcal/mol) via hydrogen bonds, π—π stacking, and π—S interactions. Enrichment analysis highlighted the response to reactive oxygen species, caveolae, endopeptidase activity, and IL-17 signaling pathway as key pathways. As revealed by in vitro experiments, omeprazole dose-dependently lessened NPCs viability (300 µM) and heightened apoptosis (28.99% apoptosis rate). Western blot showed significant upregulation of Cleaved-CASP3/pro-CASP3 ratios (P < 0.001), and immunofluorescence demonstrated CASP3 nuclear translocation in omeprazole-treated NPCs. Conclusions This study found that taking omeprazole may exacerbate IVDD, and its potential mechanism is through CASP3 leading to apoptosis of NPCs. These findings advocate cautious long-term omeprazole use in clinical practice and suggest alternative PPIs.
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spelling doaj-art-b2c0de3fde8e463eaec203eab2d8f5ab2025-08-20T02:11:22ZengBMCJournal of Orthopaedic Surgery and Research1749-799X2025-05-0120111110.1186/s13018-025-05863-4Omeprazole exacerbates intervertebral disc degeneration through Caspase-3 mediated apoptosis of nucleus pulposus cells: a Mendelian randomization, network toxicology, and in vitro experimental studyYuchao Jia0Haifan Zhao1Shengbo Huang2Baoshan Xu3Department of Minimally Invasive Spine Surgery, Tianjin HospitalDepartment of Minimally Invasive Spine Surgery, Tianjin HospitalDepartment of Minimally Invasive Spine Surgery, Tianjin HospitalDepartment of Minimally Invasive Spine Surgery, Tianjin HospitalAbstract Objective To investigate the causal correlation and toxicological mechanisms of omeprazole in intervertebral disc degeneration (IVDD), alongside a particular emphasis on Caspase-3 (CASP3) mediated apoptosis of nucleus pulposus cells (NPCs). Methods Mendelian randomization (MR): GWAS data was employed to assess causal associations between proton pump inhibitors (PPIs) and IVDD. Network toxicology: Shared omeprazole-IVDD targets were identified using STRING, SwissTargetPrediction, and GeneCards databases. Functional enrichment analysis: Biological pathways were explored by employing Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Molecular docking: Omeprazole-CASP3 binding affinity was assessed by employing AutoDock Vina. Experimental validation: Rat NPCs were subjected to CCK-8 assay viability, flow cytometry apoptosis, Western blot, and immunofluorescence. Results MR analysis suggested omeprazole substantially augmented IVDD risk (OR = 1.058, 95% CI = 1.004–1.115, P = 0.034), with no association observed for esomeprazole or lansoprazole. Network toxicology identified 11 overlapping targets, with CASP3 as the hub gene. Molecular docking revealed strong omeprazole-CASP3 binding (free energy: − 6.725 kcal/mol) via hydrogen bonds, π—π stacking, and π—S interactions. Enrichment analysis highlighted the response to reactive oxygen species, caveolae, endopeptidase activity, and IL-17 signaling pathway as key pathways. As revealed by in vitro experiments, omeprazole dose-dependently lessened NPCs viability (300 µM) and heightened apoptosis (28.99% apoptosis rate). Western blot showed significant upregulation of Cleaved-CASP3/pro-CASP3 ratios (P < 0.001), and immunofluorescence demonstrated CASP3 nuclear translocation in omeprazole-treated NPCs. Conclusions This study found that taking omeprazole may exacerbate IVDD, and its potential mechanism is through CASP3 leading to apoptosis of NPCs. These findings advocate cautious long-term omeprazole use in clinical practice and suggest alternative PPIs.https://doi.org/10.1186/s13018-025-05863-4OmeprazoleIntervertebral Disc DegenerationMendelian RandomizationNetwork ToxicologyCaspase-3
spellingShingle Yuchao Jia
Haifan Zhao
Shengbo Huang
Baoshan Xu
Omeprazole exacerbates intervertebral disc degeneration through Caspase-3 mediated apoptosis of nucleus pulposus cells: a Mendelian randomization, network toxicology, and in vitro experimental study
Journal of Orthopaedic Surgery and Research
Omeprazole
Intervertebral Disc Degeneration
Mendelian Randomization
Network Toxicology
Caspase-3
title Omeprazole exacerbates intervertebral disc degeneration through Caspase-3 mediated apoptosis of nucleus pulposus cells: a Mendelian randomization, network toxicology, and in vitro experimental study
title_full Omeprazole exacerbates intervertebral disc degeneration through Caspase-3 mediated apoptosis of nucleus pulposus cells: a Mendelian randomization, network toxicology, and in vitro experimental study
title_fullStr Omeprazole exacerbates intervertebral disc degeneration through Caspase-3 mediated apoptosis of nucleus pulposus cells: a Mendelian randomization, network toxicology, and in vitro experimental study
title_full_unstemmed Omeprazole exacerbates intervertebral disc degeneration through Caspase-3 mediated apoptosis of nucleus pulposus cells: a Mendelian randomization, network toxicology, and in vitro experimental study
title_short Omeprazole exacerbates intervertebral disc degeneration through Caspase-3 mediated apoptosis of nucleus pulposus cells: a Mendelian randomization, network toxicology, and in vitro experimental study
title_sort omeprazole exacerbates intervertebral disc degeneration through caspase 3 mediated apoptosis of nucleus pulposus cells a mendelian randomization network toxicology and in vitro experimental study
topic Omeprazole
Intervertebral Disc Degeneration
Mendelian Randomization
Network Toxicology
Caspase-3
url https://doi.org/10.1186/s13018-025-05863-4
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