Research Paper Volume 16, Issue 4 pp 3955—3972
Protective effects of triptolide against oxidative stress in retinal pigment epithelium cells via the PI3K/AKT/Nrf2 pathway: a network pharmacological method and experimental validation
- 1 Department of Ophthalmology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
- 2 Chongqing Key Laboratory of Ophthalmology, Chongqing Eye Institute, Chongqing 400016, China
Received: September 8, 2023 Accepted: January 11, 2024 Published: February 21, 2024
https://doi.org/10.18632/aging.205570How to Cite
Copyright: © 2024 Pan et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Purpose: Among aging adults, age-related macular degeneration (AMD), is a prevalent cause of blindness. Nevertheless, its progression may be halted by antioxidation in retinal pigment epithelium (RPE). The primary effective constituent of Tripterygium wilfordii Hook. F., triptolide (TP), has demonstrated anti-inflammatory, antiproliferative, and antioxidant properties. The mechanics of the protective effect of triptolide against the oxidative damage in retinal pigment epithelial (RPE) were assessed in this study.
Methods: ARPE-19 cells were pretreated with TP, and then exposed to sodium iodate (SI). First, cell viability was assessed using CCK-8. Subsequently, we measured indicators for cell oxidation including reactive oxygen species (ROS), catalase (CAT), superoxide dismutase (SOD), and malondialdehyde (MDA). Then, we used network pharmacological analysis and molecular docking to explore the signaling pathway of TP. Last, we used western blot, ELISA, and immunofluorescence assays to clarify the potential mechanistic pathways.
Results: The network pharmacology data suggested that TP may inhibit AMD by regulating the PI3K/Akt signaling pathway. Experimental results showed that the potential mechanism is that it regulates the PI3K/Akt pathway and promotes Nrf2 phosphorylation and activation, thereby raising the level of antioxidant factors (HO-1, NQO1) and reducing the generation of ROS, which inhibit oxidative damage.
Conclusion: Our findings suggested that the effect of TP on SI-exposed RPE cells principally relies on the regulation of oxidative stress through the PI3K/Akt/Nrf2 signaling pathway.
Abbreviations
GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; DHCA: Deep hypothermic circulatory arrest; DOMS: Delayed-onset muscle soreness; IV: Intravenous therapy; LD: Lethal dose; GST: Glutathione-S-transferase; NQO1: NADH Quinone oxidoreductase 1; HO-1: Heme oxygenase 1; CTD: Comparative Toxicogenomics Database; OMIM: Online Mendelian Inheritance in Man; RIPA: Radio immunoprecipitation assay; TBST: Tris-Buffered saline and tween.