BACKGROUND: Chronic Kidney Disease (CKD) is a progressive condition characterized by a decline in kidney function and structural damage, often leading to severe health outcomes. The kidneys are crucial in maintaining homeostasis, including fluid and electrolyte balance, acid-base regulation, and waste excretion. CKD affects millions worldwide, with many individuals unaware of their condition due to subtle early symptoms. Current treatments primarily delay disease progression, highlighting the need for innovative therapeutic strategies. Programmed cell death (PCD), involving mechanisms such as apoptosis and necroptosis, significantly contributes to CKD progression.
PURPOSE: Traditional Chinese Medicine (TCM), particularly Zhenwu Decoction, offers a multi-target approach that may alleviate CKD symptoms. The current study was designed to elucidate the mechanisms by which Zhenwu Decoction affects CKD progression and to validate its potential as a novel therapeutic option using bioinformatics and network pharmacology.
MATERIALS AND METHODS: Zhenwu Decoction, composed of aconite, Poria, Atractylodes macrocephala, white peony root, and ginger, was prepared by dissolving the granules in distilled water. The decoction was administered to rats at a specified dosage based on body weight. Aconite and the pharmaceutical telmisartan were also prepared and administered to evaluate their effects comparatively. The study utilized 40 Sprague-Dawley rats, divided equally by sex, to model doxorubicin-induced nephropathy. Rats were housed under controlled conditions, with daily monitoring of their health and behavior. Rats received a single tail vein injection of doxorubicin to induce nephropathy, with a control group receiving saline. Successful modeling was confirmed by observing physical symptoms and elevated urine protein levels. Rats were treated with Zhenwu Decoction, aconite, or telmisartan. The effects of these treatments on nephropathy symptoms and kidney function were assessed. Kidney tissues were analyzed using Hematoxylin and Eosin (HE) staining for morphology, Masson's Trichrome staining for fibrosis, and PASM staining for structural visualization. The study included analyzing the expression of the rapamycin target protein mTOR, with protein extraction performed on kidney samples post-treatment.
RESULTS: Our results show that Zhenwu Decoction (ZWD) effectively treats CKD by reducing symptoms, proteinuria, and kidney damage. Its efficacy stems from a multi-targeted approach, simultaneously modulating inflammation, fibrosis, and programmed cell death (PCD). ZWD significantly reduces fibrotic markers like TIMP3 and Col-IV, influencing ECM-receptor interaction and TGF-β signaling. It also modulates inflammation through the NF-$\kappa$B pathway and mTOR/autophagy axis, downregulating pro-inflammatory cytokines while increasing anti-inflammatory ones. Furthermore, ZWD regulates PCD by decreasing necroptosis markers and inhibiting ferroptosis via ALOX12 reduction, while upregulating anti-apoptotic PARP2 for enhanced DNA repair. This comprehensive action across various pathways provides superior renal protection. Despite the documented efficacy of Zhenwu Decoction (ZWD) in clinical practice, a significant gap remains in systematically integrating bioinformatics predictions with experimental validation to elucidate its multi-target mechanisms against programmed cell death (PCD) in CKD. This study was therefore designed to bridge this gap by employing a comprehensive network pharmacology approach alongside in vivo validation to definitively map ZWD's reno-protective actions onto specific inflammatory, fibrotic, and PCD pathways, thereby providing a mechanistic foundation for its therapeutic application.
CONCLUSIONS: Our present study shows the potential of Zhenwu Decoction in the management of chronic kidney disease using a bioinformatics and network pharmacology strategy to target kidney function and programmed cell death. Its active components advanced our knowledge of TCM's multi-target effects by improving kidney health in a rat model of doxorubicin-induced nephropathy.
