Network pharmacology and experimental validation to elucidate the pharmacological mechanisms of OATF against kidney stones

IntroductionOrthosiphon aristatus (Blume) Miq. (OA) is widely used in folk medicine to treat kidney stones (KS). Its total flavonoids (OATF) are the primary active constituents responsible for its therapeutic effects. However, the exact mechanism of action (MOA) remains unclear. This study aimed to...

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Main Authors: Sisi Dai, Qi Fang, Hong-Yan Li, Rui Sun, Hui-Yong Zhang, Wei Wu
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Pharmacology
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Online Access:https://www.frontiersin.org/articles/10.3389/fphar.2025.1575270/full
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author Sisi Dai
Sisi Dai
Qi Fang
Hong-Yan Li
Rui Sun
Hui-Yong Zhang
Wei Wu
Wei Wu
author_facet Sisi Dai
Sisi Dai
Qi Fang
Hong-Yan Li
Rui Sun
Hui-Yong Zhang
Wei Wu
Wei Wu
author_sort Sisi Dai
collection DOAJ
description IntroductionOrthosiphon aristatus (Blume) Miq. (OA) is widely used in folk medicine to treat kidney stones (KS). Its total flavonoids (OATF) are the primary active constituents responsible for its therapeutic effects. However, the exact mechanism of action (MOA) remains unclear. This study aimed to investigate the pharmacological activity of OATF against KS and elucidate its underlying MOA.MethodsNetwork pharmacology and molecular docking were utilized to predict the potential targets and pathways of OATF. An animal model of calcium oxalate crystal deposition was created using intraperitoneal injections of ethylene glycol (EG) and ammonium chloride (AC), alongside a model using human renal tubular epithelial cells (HK-2) induced by supersaturated oxalate (Ox) to investigate the pharmacological mechanisms of OATF against oxidative stress and apoptosis. The effects of OATF on crystal deposition and renal damage were assessed using hematoxylin-eosin (H&E) and periodic acid-Schiff (PAS) staining. Renal tubular damage and apoptosis were evaluated via TUNEL staining. The MOA was explored using Western blotting analyses.ResultsNetwork pharmacological analysis identified the EGFR/PI3K/AKT pathway as a key mechanism in KS. In vitro experimental results demonstrated that OATF effectively protected HK-2 cells from oxidative stress, inhibited calcium oxalate crystal adhesion, and reduced apoptosis. In vivo, OATF significantly decreased serum creatinine (SCR), serum calcium (Ca), serum phosphorus (P), and blood urea nitrogen (BUN) levels in CaOx-induced Sprague-Dawley (SD) rats, indicating its protective effects against KS.ConclusionOATF effectively inhibited kidney stone formation and mitigated renal injury by attenuating oxidative stress and apoptosis through activation of the EGFR/PI3K/AKT pathway. These findings highlight OATF’s therapeutic potential for KS management and provide a scientific basis for its traditional use in herbal medicine.
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spelling doaj-art-ca94e839a24b4832b3f81f8eeff2c89e2025-08-20T02:15:24ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122025-05-011610.3389/fphar.2025.15752701575270Network pharmacology and experimental validation to elucidate the pharmacological mechanisms of OATF against kidney stonesSisi Dai0Sisi Dai1Qi Fang2Hong-Yan Li3Rui Sun4Hui-Yong Zhang5Wei Wu6Wei Wu7School of Life Sciences, Zhuhai College of Science and Technology, Zhuhai, ChinaCollege of Life Sciences, Jilin University, Changchun, ChinaKey Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications, Liaoning University of Traditional Chinese Medicine, Shenyang, ChinaKey Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications, Liaoning University of Traditional Chinese Medicine, Shenyang, ChinaKey Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications, Liaoning University of Traditional Chinese Medicine, Shenyang, ChinaDepartment of Traditional Chinese Medicine, The Forth Affiliated Hospital of China Medical University, Shenyang, ChinaSchool of Life Sciences, Zhuhai College of Science and Technology, Zhuhai, ChinaSchool of Basic Medical Sciences, Peking University Health Science Center, Beijing, ChinaIntroductionOrthosiphon aristatus (Blume) Miq. (OA) is widely used in folk medicine to treat kidney stones (KS). Its total flavonoids (OATF) are the primary active constituents responsible for its therapeutic effects. However, the exact mechanism of action (MOA) remains unclear. This study aimed to investigate the pharmacological activity of OATF against KS and elucidate its underlying MOA.MethodsNetwork pharmacology and molecular docking were utilized to predict the potential targets and pathways of OATF. An animal model of calcium oxalate crystal deposition was created using intraperitoneal injections of ethylene glycol (EG) and ammonium chloride (AC), alongside a model using human renal tubular epithelial cells (HK-2) induced by supersaturated oxalate (Ox) to investigate the pharmacological mechanisms of OATF against oxidative stress and apoptosis. The effects of OATF on crystal deposition and renal damage were assessed using hematoxylin-eosin (H&E) and periodic acid-Schiff (PAS) staining. Renal tubular damage and apoptosis were evaluated via TUNEL staining. The MOA was explored using Western blotting analyses.ResultsNetwork pharmacological analysis identified the EGFR/PI3K/AKT pathway as a key mechanism in KS. In vitro experimental results demonstrated that OATF effectively protected HK-2 cells from oxidative stress, inhibited calcium oxalate crystal adhesion, and reduced apoptosis. In vivo, OATF significantly decreased serum creatinine (SCR), serum calcium (Ca), serum phosphorus (P), and blood urea nitrogen (BUN) levels in CaOx-induced Sprague-Dawley (SD) rats, indicating its protective effects against KS.ConclusionOATF effectively inhibited kidney stone formation and mitigated renal injury by attenuating oxidative stress and apoptosis through activation of the EGFR/PI3K/AKT pathway. These findings highlight OATF’s therapeutic potential for KS management and provide a scientific basis for its traditional use in herbal medicine.https://www.frontiersin.org/articles/10.3389/fphar.2025.1575270/fullOrthosiphon aristatuskidney stoneapoptosisnetwork pharmacologyEGFR/PI3K/Akt
spellingShingle Sisi Dai
Sisi Dai
Qi Fang
Hong-Yan Li
Rui Sun
Hui-Yong Zhang
Wei Wu
Wei Wu
Network pharmacology and experimental validation to elucidate the pharmacological mechanisms of OATF against kidney stones
Frontiers in Pharmacology
Orthosiphon aristatus
kidney stone
apoptosis
network pharmacology
EGFR/PI3K/Akt
title Network pharmacology and experimental validation to elucidate the pharmacological mechanisms of OATF against kidney stones
title_full Network pharmacology and experimental validation to elucidate the pharmacological mechanisms of OATF against kidney stones
title_fullStr Network pharmacology and experimental validation to elucidate the pharmacological mechanisms of OATF against kidney stones
title_full_unstemmed Network pharmacology and experimental validation to elucidate the pharmacological mechanisms of OATF against kidney stones
title_short Network pharmacology and experimental validation to elucidate the pharmacological mechanisms of OATF against kidney stones
title_sort network pharmacology and experimental validation to elucidate the pharmacological mechanisms of oatf against kidney stones
topic Orthosiphon aristatus
kidney stone
apoptosis
network pharmacology
EGFR/PI3K/Akt
url https://www.frontiersin.org/articles/10.3389/fphar.2025.1575270/full
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