Ancient Chinese Herbal Formula Decoded: How Danggui Buxue Decoction Fights Chronic Kidney Disease

An 800-year-old Chinese herbal remedy has been put under the modern scientific microscope, and the results reveal a remarkably precise molecular strategy for treating one of the most common forms of chronic kidney disease. Danggui Buxue Decoction, a two-herb formula first documented during the Jin-Yuan period in Li Dongyuan’s classic text “Neiwaishang Bianhuo Lun,” has long been used in traditional Chinese medicine to tonify qi and blood. Now, a research team combining ultra-high-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS), network pharmacology, bioinformatics, and laboratory experiments has mapped out exactly how the formula combats chronic glomerulonephritis, a progressive inflammatory kidney condition that can advance to kidney fibrosis and, ultimately, kidney failure.

The study, published in the journal Immunity, Inflammation and Disease, began with a rigorous chemical inventory of the decoction, which combines Angelicae Sinensis Radix, the root of Angelica sinensis, with Astragali Radix, the root of Astragalus membranaceus, in a precise 1:5 ratio. Using a Vanquish ultra-high-performance liquid chromatograph coupled to a QExactive HFX mass spectrometer, the researchers separated compounds on a Waters HSS T3 column at 40 degrees Celsius with a formic acid-acetonitrile gradient and electrospray ionization in both positive and negative ion modes. The analysis identified 104 compounds, dominated by phenylpropanoids and polyketides, lipids and lipid-like molecules, and organoheterocyclic compounds. This chemical fingerprint became the foundation for everything that followed.

With the compound list in hand, the team turned to network pharmacology, a computational approach that treats a multi-component herbal formula as a network of interacting molecules rather than a single active ingredient. Predicting targets for the identified compounds through SwissTargetPrediction yielded 499 drug-related targets, while searching GeneCards and OMIM for chronic glomerulonephritis produced 2,821 disease targets after redundancy removal. The intersection contained 166 overlapping targets, suggesting a substantial molecular interface between the formula and the disease. Building a protein-protein interaction network in Cytoscape and ranking targets with the CytoHubba MCC algorithm highlighted ten core targets, including B-cell lymphoma-2 (Bcl-2) and Caspase-3, both central players in apoptosis, the genetically programmed form of cell death.

Enrichment analysis through the Metascape platform pointed decisively toward the NF-κB signaling pathway, a master regulator of inflammation. To test whether this computational prediction reflected real biology, the researchers mined two independent gene expression datasets from the Gene Expression Omnibus, GSE116626 with 74 chronic glomerulonephritis samples and GSE104066 with 70 samples. After batch correction and standardization, differential expression analysis identified 414 significantly upregulated genes and 379 downregulated genes, and KEGG pathway analysis showed these genes were also significantly enriched in the NF-κB signaling pathway. The convergence of network pharmacology and patient gene expression data on the same pathway gave the team strong grounds to pursue experimental validation.

Molecular docking provided a structural bridge between chemistry and biology. Five active ingredients with high network centrality, 3-Hexen-1-ol O-β-d-glucopyranoside, 4-hydroxybenzoic acid, caffeic acid, calycosin, and lucidal, were docked against Bcl-2, Caspase-3, and TLR4. All fifteen compound-target pairs showed binding energies below the -5 kcal/mol threshold generally considered indicative of strong binding. Calycosin bound Caspase-3 at -7.2 kcal/mol, while lucidal achieved -8.5 kcal/mol against Caspase-3 and -7.8 kcal/mol against Bcl-2, stabilized by multiple hydrogen bonds with key amino acid residues. These docking scores suggested the herbal compounds could physically engage the apoptotic and inflammatory machinery implicated in the disease.

The first line of experimental evidence came from cell studies using HBZY-1 rat glomerular mesangial cells, whose abnormal behavior is a central driver of chronic glomerulonephritis progression. Rather than applying crude herbal extracts, the team prepared drug-containing serum by administering DBD to rats at three doses, 1.89, 3.78, and 7.56 grams per kilogram, then collecting the resulting serum, an approach that captures only the compounds that actually enter the bloodstream after metabolism. When LPS-stimulated mesangial cells were treated with DBD-containing serum, the abnormally elevated cell viability dropped, secretion of the anti-inflammatory cytokines IL-4 and IL-10 recovered, and Transwell assays showed markedly reduced migration and invasion. Immunofluorescence confirmed that TLR4 and NF-κB protein expression, which LPS had pushed upward, fell significantly after treatment.

The in vivo experiments used a classic adriamycin-induced rat model of chronic glomerulonephritis, established through two tail-vein injections and confirmed by 24-hour urinary protein exceeding 50 milligrams per kilogram. Rats receiving DBD for 30 days showed striking improvements across the board: 24-hour urine protein, serum creatinine, and blood urea nitrogen all fell significantly, while body weight recovered and the kidney index normalized. Pro-inflammatory TNF-α and IL-6 levels declined, and the previously suppressed IL-4 and IL-10 rose. Histological staining with hematoxylin and eosin, Masson, and PAS revealed that the adriamycin-induced damage, including glomerular atrophy, basement membrane thickening, collagen fiber deposition, and inflammatory cell infiltration, was substantially reversed in the treated animals.

The mechanistic heart of the study lies in its analysis of apoptosis and the TLR4/NF-κB axis. In the diseased rats, apoptotic activity was paradoxically suppressed: Cleaved Caspase-3 staining was weak, TUNEL-positive cells were scarce, and mitochondrial membrane potential measured by JC-1 staining indicated inhibited apoptosis. Western blotting showed downregulation of the pro-apoptotic proteins Bax, cytochrome C, Caspase-9, and Caspase-3, alongside upregulation of the anti-apoptotic Bcl-2. DBD treatment reversed this pattern, restoring pro-apoptotic signaling and promoting the controlled cell death needed to clear damaged glomerular cells. Simultaneously, RT-qPCR, Western blotting, immunohistochemistry, and immunofluorescence all demonstrated that DBD downregulated TLR4, MyD88, IκBα, and NF-κB, damping the inflammatory cascade that LPS and adriamycin normally ignite.

This dual action makes biological sense in light of the disease mechanism. When TLR4 on mesangial cell surfaces is activated, it recruits MyD88, promotes IκB phosphorylation and degradation, and triggers NF-κB nuclear translocation, where the transcription factor drives expression of inflammatory cytokines such as TNF-α and IL-1β and of the anti-apoptotic protein Bcl-2. By suppressing this pathway, DBD reduces inflammatory cytokine release; by simultaneously promoting Bax-mediated cytochrome C release and Caspase-9 and Caspase-3 activation, it restores the apoptotic balance whose disruption drives both pathological mesangial accumulation and loss of kidney filtering cells. The authors note that apoptosis is a double-edged sword, but their data indicate that reactivating it in this context slows disease progression rather than accelerating tissue damage.

The findings carry practical significance beyond the laboratory. Current clinical management of chronic glomerulonephritis relies on angiotensin-converting enzyme inhibitors, corticosteroids, and immunosuppressants, all of which carry limitations including variable drug sensitivity, symptom recurrence, and long-term adverse effects such as hyperkalemia, dry cough, increased infection risk, and digestive reactions. A multi-component, multi-target herbal formula that modulates both inflammation and apoptosis could offer a gentler complementary strategy, and the study’s systematic workflow, from chemical profiling through network prediction to animal and cellular validation, provides a template for scientifically grounding other traditional formulas. The researchers conclude that DBD exerts its therapeutic effects on chronic glomerulonephritis by regulating the TLR4/NF-κB signaling pathway, promoting apoptosis, and alleviating inflammation, laying an experimental foundation for explaining its clinical efficacy in modern pharmacological terms.

Subject of Research: Mechanism of action of Danggui Buxue Decoction in treating chronic glomerulonephritis via the TLR4/NF-κB signaling pathway

Article Title: Exploring the Mechanism of Action of Danggui Buxue Decoction in Treating Chronic Glomerulonephritis Based on UPLC‐MS/MS and Network Pharmacology

Article References: Liu, Q., Wang, Y., Wang, X., Yang, H., Pan, Z., & Guo, D. (2026). Exploring the Mechanism of Action of Danggui Buxue Decoction in Treating Chronic Glomerulonephritis Based on UPLC‐MS/MS and Network Pharmacology. Immunity, Inflammation and Disease, 14(9), Article e70497. https://doi.org/10.1002/iid3.70497

Image Credits: AI Generated

DOI: 10.1002/iid3.70497

Keywords: Danggui Buxue Decoction, chronic glomerulonephritis, network pharmacology, UPLC-MS/MS, TLR4/NF-κB pathway, apoptosis, traditional Chinese medicine, kidney disease, HBZY-1 mesangial cells, molecular docking, inflammation, astragalus