Kaempferol Eases Muscle Wasting in Mice With Cancer Cachexia

Cancer cachexia, the devastating wasting syndrome that strips skeletal muscle from people with advanced cancer and is estimated to contribute to roughly 20 to 30 percent of cancer-related deaths, has few effective treatments. A new study published in Biological Procedures Online reports a data-driven route to a potential candidate: the plant flavonoid kaempferol, identified through a combination of literature mining, network pharmacology and laboratory testing in tumor-bearing mice and muscle cells. The work is strictly preclinical, and its authors are careful to note that the muscle improvements they observed cannot yet be separated from the drug’s effect on tumor growth itself.

The team began with a bibliometric sweep of the Web of Science Core Collection and Scopus databases, searching from database inception to December 31, 2025 for studies combining cachexia or muscle wasting terms with traditional Chinese medicine terms. From 4,285 retrieved records, duplicate removal, title and abstract screening and full-text assessment left 142 eligible publications. Frequency statistics identified the four most studied formulas — Liujunzi decoction, Renshen Yangrong decoction, Shiquan Dabu decoction and Buzhong Yiqi decoction — and, after decomposing formulas into single herbs, five core herbs: Panax ginseng (82 records), Glycyrrhiza uralensis (70), Atractylodes macrocephala (65), Astragalus membranaceus (55) and Poria cocos (54).

Next came network pharmacology. Chemical constituents of the five core herbs were pulled from the TCMSP database, filtered by oral bioavailability and drug-likeness thresholds, and intersected with cachexia-related disease targets from GeneCards and OMIM. A protein–protein interaction network built in STRING yielded 19 hub genes, including AKT1, TNF, IL6 and STAT3, and enrichment analysis pointed to PI3K–Akt signaling, IL-17 signaling and other inflammation- and metabolism-related pathways. Quercetin actually connected to more hub genes (17) than kaempferol (7), but the authors applied three criteria rather than network connectivity alone. Kaempferol was present in three of the five core herbs — ginseng, astragalus and licorice — versus quercetin’s two, and its reported anti-inflammatory and lipid metabolism-regulating activities matched the enriched pathways. On that combined evidence, kaempferol was chosen for experimental evaluation.

The in vivo test used male BALB/c nude mice inoculated subcutaneously with human A549 lung cancer cells. Seven days later, once mean tumor volume reached about 100 cubic millimeters, tumor-bearing mice were randomly assigned to a model group or to low-, medium- or high-dose kaempferol groups receiving 25, 50 or 100 milligrams per kilogram per day by oral gavage. Over the 21-day experiment, the researchers tracked body weight, tumor dimensions, food intake and forelimb grip strength, then collected gastrocnemius muscle for histology, western blotting, immunofluorescence, electron microscopy and untargeted metabolomics.

The headline result concerned tumor burden and the wasting phenotype together. At the endpoint, mean tumor weight was 2.01 plus or minus 0.15 grams in the untreated model group but fell to 1.12 plus or minus 0.16 grams with high-dose kaempferol. That reduction was accompanied by significant improvements in daily food intake, which reached 3.62 plus or minus 0.08 grams, estimated tumor-free body weight, which reached 24.9 plus or minus 0.5 grams, gastrocnemius wet weight, which rose to 0.56 plus or minus 0.03 grams, and forelimb grip strength, which climbed to 81.9 plus or minus 2.8 grams, all compared with the model group at P < 0.05. Notably, significant gains in myofiber cross-sectional area and in the molecular markers described below appeared only at the high dose; the low and medium doses showed non-significant trends.

At the molecular level, the tumor-bearing mice showed elevated expression of Atrogin-1 and MuRF1, two muscle-enriched E3 ubiquitin ligases that drive ubiquitin–proteasome-mediated protein degradation during atrophy. High-dose kaempferol significantly reduced both the fluorescence signals and the protein levels of these ligases, consistent with the observed increases in muscle fiber size and grip strength. Serum concentrations of the inflammatory mediators TNF-α, IL-6 and prostaglandin E2 were also significantly lower after high-dose treatment, and skeletal muscle COX-2, an enzyme in the arachidonic acid to prostaglandin pathway, was reduced as well.

Untargeted metabolomics of gastrocnemius tissue, performed on six animals per group using an ultra-high-performance liquid chromatography system coupled to a high-resolution Orbitrap mass spectrometer, added a metabolic dimension. Differential metabolites screened at VIP > 1, absolute log2 fold change > 1 and P < 0.05 mapped to glycerophospholipid metabolism, arachidonic acid metabolism, linoleic acid metabolism, several amino acid pathways and autophagy-related pathways. Compared with the model group, high-dose kaempferol was associated with lower arachidonic acid abundance and higher abundance of selected phosphatidylcholine and phosphatidylethanolamine species, branched-chain amino acids, L-arginine and L-carnitine — metabolites relevant to muscle membrane organization, mitochondrial metabolism and functional maintenance.

Autophagy findings were more equivocal, and the authors handled them with unusual candor. The tumor-bearing mice showed higher LC3B-II/LC3B-I ratios, higher p62 levels and more autophagic vacuole-like structures on transmission electron microscopy, all of which decreased with high-dose kaempferol. But LC3B-II reflects autophagosome abundance at a single time point and can rise either from enhanced formation or impaired degradation, while p62 is regulated by autophagic degradation, transcription, proteasomal activity and cellular stress. The non-parallel changes in these markers after prostaglandin E2 supplementation in the cell experiments underscore the interpretive difficulty. Without dynamic flux assays or lysosomal inhibition experiments, the direction of autophagic flux remains unresolved.

To examine muscle-cell responses without the confounding influence of changing tumor mass, the team used C2C12 myotubes exposed to A549-conditioned medium. Conditioned medium reduced Myh4 mRNA and increased Atrogin-1 and MuRF1 mRNA; 25 micromolar kaempferol reversed these shifts. Adding 1 micromolar prostaglandin E2 worsened the atrophy gene pattern and raised the LC3B-II/LC3B-I ratio, and kaempferol co-treatment shifted the endpoints back. Kaempferol also reduced cytoplasmic vacuolation quantified by electron microscopy, both with and without supplemental prostaglandin E2. This pattern is consistent with a link between prostaglandin E2-related signaling and kaempferol’s molecular effects, though prostaglandin E2 was tested only in the presence of conditioned medium, without pathway inhibition or receptor-specific intervention.

The limitations matter for anyone hoping to translate these results. Because kaempferol shrank tumors and improved food intake simultaneously, the muscle improvements may partly reflect indirect systemic benefits rather than direct muscle protection; tumor burden-matched, pair-feeding and tumor-independent wasting models are needed to disentangle the two. The focused histological, molecular and ultrastructural analyses used only three biological replicates per group and should be regarded as exploratory. Adipose tissue wasting and muscle fiber-type composition were not assessed, and the cell work lacked myotube diameter measurements. The authors position kaempferol not as a ready therapy but as a candidate warranting further investigation, with the priorities being to clarify prostaglandin E2-related signaling, measure autophagic flux directly and separate muscle-specific actions from the systemic consequences of a smaller tumor.

Subject of Research: Identification and preclinical evaluation of kaempferol as a traditional Chinese medicine-derived candidate compound for cancer cachexia-associated skeletal muscle wasting

Article Title: Identification of Kaempferol for Skeletal Muscle Wasting in Cancer Cachexia Through Bibliometric Analysis and Network Pharmacology: Metabolomic and Experimental Evaluation

Article References: Zhu, C., Chen, S., Qu, Y., He, M., Nie, R., Liu, Z., Hu, Y., Huang, J., Shen, C., Chen, Z., Wen, J., Wang, L., & Sun, Y. (2026). Identification of Kaempferol for Skeletal Muscle Wasting in Cancer Cachexia Through Bibliometric Analysis and Network Pharmacology: Metabolomic and Experimental Evaluation. Biological Procedures Online, 28(1), Article 42. https://doi.org/10.1186/s12575-026-00361-1

Image Credits: AI Generated

DOI: 10.1186/s12575-026-00361-1

Keywords: cancer cachexia, kaempferol, skeletal muscle wasting, network pharmacology, bibliometric analysis, traditional Chinese medicine, metabolomics, A549 tumor-bearing mice, C2C12 myotubes, autophagy, prostaglandin E2, preclinical study