Network-Pharmacology-Guided Validation of Phyllanthus niruri Nephroprotection Against Doxorubicin: An In Vitro–In Vivo Translational Study

Abstract

Background: Doxorubicin is a cornerstone anthracycline chemotherapeutic whose clinical utility is constrained by dose-limiting nephrotoxicity, and no approved pharmacological prevention currently exists. Phyllanthus niruri L. (family Phyllanthaceae), known in Indonesia as meniran, is a phytotherapeutic herb whose constituents (phyllanthin, hypophyllanthin, quercetin, rutin, gallic acid, and corilagin) target the oxidative-stress and inflammatory pathways implicated in doxorubicin renal injury.

Objective: To test whether standardised P. niruri ethanolic extract attenuates doxorubicin-induced nephrotoxicity through a mechanistically coherent multi-target pathway, using a tiered translational design.

Methods: Network-pharmacology discovery using SwissTargetPrediction, the Traditional Chinese Medicine Systems Pharmacology database (TCMSP), GeneCards, and Online Mendelian Inheritance in Man (OMIM) intersected six P. niruri active compounds with 842 nephrotoxicity-associated genes. In vitro validation used HK-2 human proximal tubular cells exposed to doxorubicin (2 µM, 24 h) with or without standardised P. niruri 70% ethanolic extract (25, 50, and 100 µg/mL). In vivo validation used 48 male Sprague–Dawley rats (eight per arm) given a single intraperitoneal dose of doxorubicin (15 mg/kg) and oral P. niruri at 100, 200, or 400 mg/kg/day for 14 days.

Results: Six hub targets (TNF, IL6, NFKB1, CASP3, BAX, and BCL2) and four enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways (TNF signalling, MAPK signalling, PI3K/Akt signalling, and apoptosis) were identified. The P. niruri extract produced a dose-dependent rescue of HK-2 viability (84.6% at 100 µg/mL versus 41.8% in doxorubicin-only controls; p < 0.001). At 400 mg/kg, it reduced serum creatinine, blood urea nitrogen, urinary kidney injury molecule-1 (KIM-1), and neutrophil gelatinase-associated lipocalin (NGAL) by 55–70%, with concordant restoration of glutathione and superoxide dismutase. The network-pharmacology hub targets showed transcriptionally coherent modulation.

Conclusion: Standardised P. niruri ethanolic extract is a mechanism-anchored herbal candidate for the adjunctive prevention of doxorubicin-induced kidney injury, warranting clinical translation.