Prenylated Stilbenoids Confer Sex-Dependent Cardioprotection in Rats

Introduction: Cardiovascular diseases (CVD) remain the leading cause of mortality and hypothyroidism is a major risk factor. While stilbenoids such as resveratrol possess cardioprotective properties, prenylated stilbenoids such as arachidin-3 (Ara-3) are considered more bioavailable. However, their in vivo therapeutic potential and the underlying molecular mechanisms remain unclear. It was hypothesized that Ara-rich extracts derived from elicited peanut hairy root cultures confer cardioprotection via transcriptional and anti-inflammatory effects. Methods: A well-established propylthiouracil-based diet-induced hypothyroid cardiomyopathic cardiac dysfunction (CCD) rat model was used. Young adult Sprague-Dawley rats (n=10; males=5; females=5) were divided into Control+Vehicle, CCD+Vehicle, CCD+resveratrol (1.5 mg/kg/day), CCD+Ara-3 (1.5 mg/kg/day), CCD+Extract-20 mg/kg/day (E-20) and CCD +Extract-40 mg/kg/day (E-40) groups for a treatment duration of 6 weeks. Morphometric, cardiac ultrasound, hematological, and plasma lipid parameters were assessed. Then mRNA sequencing and inflammatory cytokine profiling were performed. Statistical analyses were done using One-Way Analysis of Variance followed by Tukey’s Post Hoc analyses and T-tests using GraphPad Prism and tools such as R for RNA-seq analyses. Results: The prenylated stilbenoids were isolated from elicited peanut hairy roots and purified and analyzed using high-performance liquid chromatography. Based on organ morphometrics, hematological and lipid parameters, both Ara-rich extracts and Ara-3 were non-toxic and safe. Cardiac ultrasound echocardiography showed significant improvements in percent changes of left ventricular (LV) fractional shortening in females following treatment compared to CCD, i.e., 32.6% (p<0.01), 21.8%, 24.2%, 25.4% (p<0.05) in E-40, Ara-3, E-20, and resveratrol, respectively. In males, treatment compared to CCD improved by 15.8% (p<0.05), 12.2%, 7.8%, and 9% in Ex-40, Ara-3, E-20, and resveratrol, respectively. The transcriptome analyses identified significant differentially expressed mRNAs in all treatment groups in both males and females. Kyoto Encyclopedia of Genes and Genomes, KEGG analyses of CCD vs CCD+E-40 showed significant enrichments in oxidative phosphorylation, and ribosomal pathway genes in both males and females. Females also showed enrichment in Parkinson’s and Alzheimer’s pathways. In CCD vs CCD+Ara-3, KEGG enriched drug and xenobiotic metabolism by cytochrome P450. Gene Ontology, GO analyses in CCD vs CCD+E-40 showed significant enrichment in mitochondrial genes in females and protein and peptide synthesis genes in males. Furthermore, cardiac tissue inflammatory profiling showed significant improvements in targets including Jam-A and RANTES. Conclusion: TheE-40 and Ara-3 significantly improved cardiac functional, transcriptional, and inflammatory profile in cardiomyopathic cardiac dysfunction in vivo. This is the first study known to comprehensively demonstrate the safe and cardioprotective effects of prenylated stilbenoids. Wethank Arkansas Biosciences Institute for funding both Dr. Fabricio Medina-Bolivar and Dr. Viswanathan Rajagopalan on this project. This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.