Exploration of the Effect and Mechanism of Fructus Lycii, Rehmanniae Radix Praeparata, and Paeonia lactiflora in the Treatment of AMD Based on Network Pharmacology and in vitro Experimental Verification

Abstract

Purpose The aim of this study was to observe the mechanism of Fructus Lycii (FL), Rehmanniae Radix Praeparata (RRP) and Paeonia lactiflora (PL) in treating age-related macular degeneration (AMD) based on network pharmacology and biological experiments. Methods Bioactive compounds, potential targets of FL, RRP and PL, and genes related to AMD, were acquired from public databases. Functional and pathway enrichment analyses of the core targets were conducted by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Subsequently, the finding was further verified with cell experiments. The MTT assay and flow cytometric analysis were used to assess cell viability and apoptosis. The production of reactive oxygen species (ROS) was analyzed by DCFH-DA staining; the activity of antioxidant enzymes was chemically measured with assay kits. The expression of key proteins was evaluated by Western blot analysis. Results Fifty-nine active compounds, 182 potential targets, and 2536 AMD-related human genes were identified. A total of 103 key targets of the three herbs on AMD were identified by protein–protein interaction (PPI) analysis. The abovementioned targets were correlated with nuclear receptor activity, oxidative stress, and apoptosis pathways according to the GO and KEGG analyses. MTT assay and flow cytometry demonstrated that pretreatment of ARPE-19 cells with the three herbs significantly increased cell viability and decreased apoptosis induced by H 2 O 2 . The three herbs might reduce the intracellular ROS levels and increase the SOD and CAT activities after H 2 O 2 . Furthermore, the three herbs significantly inhibited oxidative stress via increasing the expression of Nrf2, HO-1 and NQO1. Conclusion The combined results of network pharmacology and validation experiments showed that FL, RRP and PL reduce oxidative stress and apoptosis in RPE cells to exert its effect in the treatment of AMD.

The synergistic effect of Angelica sinensis (Oliv.) Diels and Rehmannia glutinosa (Gaertn.) DC . on antioxidant activity and protective ability against cell injury

“…SHSY-5Y cells pretreated with trans-ferulic acid were able to alleviate cell death caused by H 2 O 2 -induced toxicity as well as decreased ROS level and apoptotic cells (Gay et al, 2018). The pretreatment of ARPE-19 cells with SDH significantly increased cell viability, decreased intracellular ROS levels, and induced cell apoptosis by H 2 O 2 (Chen et al, 2021).…”

“…SHSY‐5Y cells pretreated with trans‐ferulic acid were able to alleviate cell death caused by H 2 O 2 ‐induced toxicity as well as decreased ROS level and apoptotic cells (Gay et al, 2018). The pretreatment of ARPE‐19 cells with SDH significantly increased cell viability, decreased intracellular ROS levels, and induced cell apoptosis by H 2 O 2 (Chen et al, 2021). Catalpol, an active component of SDH, had neuroprotective properties and improvement in learning and memory disorders by elevating brain‐derived neurotrophic factor and attenuating neuronal apoptosis and energy metabolism failure (Yuan et al, 2018).…”

Poria acid alleviates oxidative damage in mice with dry age-related macular degeneration by activating the Nrf2/HO-1 signaling pathway

“…Nrf2 can be found in the cytoplasm as a dimer formed with Kelch-like ECH-associated protein 1 (Keap1), however, when subjected to stimuli such as oxidative stress, it will dissociate from Keap1 and transferred to the nucleus where it binds to ARE, and regulate the expression of ARE-mediated phase II detoxi cation and antioxidant enzymes, including heme oxygenase-1 (HO-1), glutamate-Cysteine ligase (GCL) and NAD(P)H: quinone oxidoreductase (NQO1) (9), among which, HO-1 showed to be an important antioxidant enzyme that reduces cell damage (10). The upregulation of Nrf2 protects retinal cells from oxidative damage and rescues the function of the retina (11)(12)(13) Recent studies have demonstrated, the Nrf2/HO-1 signaling pathway can be activated by abundant bioactive compounds from natural products to regulate the traumatic and stressed status of the retina (14).…”

Ming-Mu-Di-Huang-Pill Activates SQSTM1 via AMPK-Mediated Autophagic KEAP1 Degradation and Protects RPE Cells from Oxidative Damage

“…According to TCM theory, the MMDH pill contains 11 herbs, possessing the functions of nourishing the kidney, calming the liver, and improving eyesight. Our previous study indicated that Fructus lycii, Rehmanniae Radix Praeparata, and Paeonia lactiflora (the main three herbs of the MMDH pill) reduce H 2 O 2 -induced oxidative stress and apoptosis in RPE cells [ 16 ].…”