Indole-3-Acetic Acid: Promising Protective Agent Against Methotrexate-Induced Liver Injury via Modulation of TLR4/NF-κB/Caspase-3 Pathway

<b>Background and Purpose:</b> Methotrexate (MTX) is a widely used therapeutic agent for inflammatory and malignant diseases; however, its prolonged use is associated with hepatotoxicity through mechanisms that remain inadequately understood. This study aims to elucidate these mechanisms...

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Main Authors: Sumayya A. Alturaif, Ahlam Alhusaini, Wedad Sarawi, Iman Hasan, Juman Alsaab, Rehab Ali, Raeesa Mohammed, Sahar S. Alotaibi, Faris Almutairi, Shaikha Alsaif, Ebtesam Alsultan, Ebtesam Aljasas, Sary Alsanea
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Language:English
Published: MDPI AG 2025-06-01
Series:Pharmaceuticals
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Online Access:https://www.mdpi.com/1424-8247/18/6/828
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author Sumayya A. Alturaif
Ahlam Alhusaini
Wedad Sarawi
Iman Hasan
Juman Alsaab
Rehab Ali
Raeesa Mohammed
Sahar S. Alotaibi
Faris Almutairi
Shaikha Alsaif
Ebtesam Alsultan
Ebtesam Aljasas
Sary Alsanea
author_facet Sumayya A. Alturaif
Ahlam Alhusaini
Wedad Sarawi
Iman Hasan
Juman Alsaab
Rehab Ali
Raeesa Mohammed
Sahar S. Alotaibi
Faris Almutairi
Shaikha Alsaif
Ebtesam Alsultan
Ebtesam Aljasas
Sary Alsanea
author_sort Sumayya A. Alturaif
collection DOAJ
description <b>Background and Purpose:</b> Methotrexate (MTX) is a widely used therapeutic agent for inflammatory and malignant diseases; however, its prolonged use is associated with hepatotoxicity through mechanisms that remain inadequately understood. This study aims to elucidate these mechanisms and assess the hepatoprotective potential of indole-3-acetic acid (IAA). <b>Methods:</b> Rats were allocated into five groups: control (group 1), IAA-treated (group 2), MTX-treated (group 3), quercetin (QUR) + MTX (group 4), and IAA + MTX (group 5). Hepatic function was assessed through the evaluation of serum liver enzymes, oxidative stress, and inflammatory and apoptotic markers using biochemical, molecular, histopathological, and immunohistochemical analyses. <b>Results:</b> The MTX-treated group demonstrated a significant increase in hepatic oxidative stress, inflammation, and apoptotic markers. Co-administration of IAA or QUR with MTX markedly reduced malondialdehyde (MDA) levels, while enhancing glutathione (GSH) levels and superoxide dismutase (SOD) activity. Moreover, hepatic inflammatory markers, including TNF-α, IL-6, and IL-1β, were significantly decreased in the IAA- and QUR-treated groups. Immunohistochemical analysis further revealed a reduced expression of NF-κB, TLR4, and caspase-3 in hepatic tissues following QUR-MTX or IAA-MTX treatments. <b>Conclusions:</b> IAA exhibited hepatoprotective effects against MTX-induced liver injury, comparable to QUR, by modulating the TLR4/NF-κB/caspase-3 pathway. These findings highlight its potential clinical application in reducing MTX-associated hepatic complications.
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spelling doaj-art-d948669a304c441d8c63dcd789ff32f92025-08-20T03:29:48ZengMDPI AGPharmaceuticals1424-82472025-06-0118682810.3390/ph18060828Indole-3-Acetic Acid: Promising Protective Agent Against Methotrexate-Induced Liver Injury via Modulation of TLR4/NF-κB/Caspase-3 PathwaySumayya A. Alturaif0Ahlam Alhusaini1Wedad Sarawi2Iman Hasan3Juman Alsaab4Rehab Ali5Raeesa Mohammed6Sahar S. Alotaibi7Faris Almutairi8Shaikha Alsaif9Ebtesam Alsultan10Ebtesam Aljasas11Sary Alsanea12Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi ArabiaDepartment of Pharmacology and Toxicology, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi ArabiaDepartment of Pharmacology and Toxicology, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi ArabiaDepartment of Pharmacology and Toxicology, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi ArabiaDepartment of Pharmacology and Toxicology, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi ArabiaDepartment of Pharmacology and Toxicology, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi ArabiaDepartment of Anatomy, College of Medicine, King Saud University, P.O. Box 2925, Riyadh 11461, Saudi ArabiaDepartment of Pharmacology and Toxicology, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi ArabiaDepartment of Pharmacology and Toxicology, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi ArabiaDepartment of Training Unit, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi ArabiaDepartment of Pharmacognosy, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi ArabiaDepartment of Pharmacognosy, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi ArabiaDepartment of Pharmacology and Toxicology, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia<b>Background and Purpose:</b> Methotrexate (MTX) is a widely used therapeutic agent for inflammatory and malignant diseases; however, its prolonged use is associated with hepatotoxicity through mechanisms that remain inadequately understood. This study aims to elucidate these mechanisms and assess the hepatoprotective potential of indole-3-acetic acid (IAA). <b>Methods:</b> Rats were allocated into five groups: control (group 1), IAA-treated (group 2), MTX-treated (group 3), quercetin (QUR) + MTX (group 4), and IAA + MTX (group 5). Hepatic function was assessed through the evaluation of serum liver enzymes, oxidative stress, and inflammatory and apoptotic markers using biochemical, molecular, histopathological, and immunohistochemical analyses. <b>Results:</b> The MTX-treated group demonstrated a significant increase in hepatic oxidative stress, inflammation, and apoptotic markers. Co-administration of IAA or QUR with MTX markedly reduced malondialdehyde (MDA) levels, while enhancing glutathione (GSH) levels and superoxide dismutase (SOD) activity. Moreover, hepatic inflammatory markers, including TNF-α, IL-6, and IL-1β, were significantly decreased in the IAA- and QUR-treated groups. Immunohistochemical analysis further revealed a reduced expression of NF-κB, TLR4, and caspase-3 in hepatic tissues following QUR-MTX or IAA-MTX treatments. <b>Conclusions:</b> IAA exhibited hepatoprotective effects against MTX-induced liver injury, comparable to QUR, by modulating the TLR4/NF-κB/caspase-3 pathway. These findings highlight its potential clinical application in reducing MTX-associated hepatic complications.https://www.mdpi.com/1424-8247/18/6/828methotrexateindole 3-acetic acidoxidative stressinflammationapoptosis
spellingShingle Sumayya A. Alturaif
Ahlam Alhusaini
Wedad Sarawi
Iman Hasan
Juman Alsaab
Rehab Ali
Raeesa Mohammed
Sahar S. Alotaibi
Faris Almutairi
Shaikha Alsaif
Ebtesam Alsultan
Ebtesam Aljasas
Sary Alsanea
Indole-3-Acetic Acid: Promising Protective Agent Against Methotrexate-Induced Liver Injury via Modulation of TLR4/NF-κB/Caspase-3 Pathway
Pharmaceuticals
methotrexate
indole 3-acetic acid
oxidative stress
inflammation
apoptosis
title Indole-3-Acetic Acid: Promising Protective Agent Against Methotrexate-Induced Liver Injury via Modulation of TLR4/NF-κB/Caspase-3 Pathway
title_full Indole-3-Acetic Acid: Promising Protective Agent Against Methotrexate-Induced Liver Injury via Modulation of TLR4/NF-κB/Caspase-3 Pathway
title_fullStr Indole-3-Acetic Acid: Promising Protective Agent Against Methotrexate-Induced Liver Injury via Modulation of TLR4/NF-κB/Caspase-3 Pathway
title_full_unstemmed Indole-3-Acetic Acid: Promising Protective Agent Against Methotrexate-Induced Liver Injury via Modulation of TLR4/NF-κB/Caspase-3 Pathway
title_short Indole-3-Acetic Acid: Promising Protective Agent Against Methotrexate-Induced Liver Injury via Modulation of TLR4/NF-κB/Caspase-3 Pathway
title_sort indole 3 acetic acid promising protective agent against methotrexate induced liver injury via modulation of tlr4 nf κb caspase 3 pathway
topic methotrexate
indole 3-acetic acid
oxidative stress
inflammation
apoptosis
url https://www.mdpi.com/1424-8247/18/6/828
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