Periplocymarin is an effective component of Periplocae Cortex, which was wildly used as an ingredient in Traditional Chinese Medicine. Our group previously reported that periplocymarin exerted cardiotonic role via promoting calcium influx. However, its exact role in the pathogenesis of myocardial fibrosis has not been elucidated yet. The present study was aimed at determining the potential effect and underlying mechanism of periplocymarin in isoproterenol (ISO)-induced myocardial fibrosis. C57BL/6 mice were subcutaneously injected with ISO (5 mg/kg/day) or saline for 1 week. The early-to-atrial wave ratio (E/A ratio) measured by echocardiography revealed that ISO-induced heart stiffness was remarkably reversed by administration of periplocymarin (5 mg/kg/day). Masson trichrome staining exhibited that treatment of periplocymarin reduced the excessive deposition of extracellular matrix (ECM). Further investigations employing real-time PCR and western blot demonstrated that periplocymarin suppressed the expression of fibrosis related genes (Col1a1, Col3a1, Acta2 and Tgfb1) and proteins (Collagen I, Collagen III, α-SMA and TGF-β1) induced by ISO. Metabolomics analysis demonstrated that periplocymarin ameliorated the disorders triggered by ISO and many of the differential metabolic substances were involved in amino acid, glucose and lipid metabolism. Further analysis using network pharmacology revealed that three key genes, namely NOS2, NOS3 and Ptgs2, may be the potential targets of periplocymarin and responsible for the disorders. Validation using heart tissues showed that the mRNA expression of NOS3 was decreased while Ptgs2 was increased upon ISO treatment, which were reversed by periplocymarin. Moreover, the expression of COX-2 (Ptgs2 encoded protein) was consistent with the aspect of Ptgs2 mRNA, while eNOS (NOS3 encoded protein) expression was unchanged. In vitro studies exhibited that periplocymarin exerts anti-fibrotic function via regulating at least eNOS and COX-2 in cardiomyocyte. Taken together, periplocymarin protects against myocardial fibrosis induced by β-adrenergic activation, the potential mechanism was that periplocymarin targeted on, at least eNOS and COX-2, to improve the metabolic processes of cardiomyocyte and thus attenuated the myocardial fibrosis. Our study highlighted that periplocymarin is a potential therapeutic agent for the prevention of myocardial fibrosis.

Background: Intestinal epithelial barrier (IEB) dysfunction is related to multiple gastrointestinal disorders, notably inflammatory bowel disease (IBD). Betulinic acid (BA), a compound derived from birch bark, has demonstrated potential therapeutic benefits in IBD. Nevertheless, the impact of BA on IEB function has not been fully elucidated. Methods: The current study aimed to explore the potential underlying mechanisms of BA in dextran sodium sulfate (DSS)-induced IBD in mice and co-culture models involving Caco-2/HT29-MTX-E12 cell monolayers or mouse intestinal organoids (IOs) in conjunction with macrophages stimulated by lipopolysaccharide (LPS). Results: In vivo, BA treatment significantly improved body weight and colon length, alleviated disease activity index (DAI) scores, and reduced colonic histopathological injury in IBD mice. In vitro, BA reduced the flux of FITC-dextran; increased the TEER; and decreased the production of IL-6, IL-1β, and TNF-α while increasing IL-10 mRNA levels. Additionally, BA enhanced IEB formation by upregulating ZO-1, occludin (OCLN), and claudin-1 (CLDN1). Molecular docking studies revealed significant docking scores and interactions between BA and PPAR-γ. Moreover, BA significantly upregulated PPAR-γ protein expression, decreased NF-κB and MLC2 phosphorylation, and reduced MLCK protein expression. However, this effect was reversed by GW9662, an effective PPAR-γ antagonist. Conclusions: The findings reveal that BA mitigates IBD by safeguarding the intestinal barrier against dysfunction. This effect may be attributed to its ability to suppress inflammation and enhance the expression of tight junction proteins by modulating the PPAR-γ/NF-κB signaling pathway.

Periplocymarin, which belongs to cardiac glycosides, is an effective component extracted from Periplocae Cortex. However, its cardiovascular effects remain unidentified. In the present study, injection of periplocymarin (5 mg/kg) through external jugular vein immediately increased the mean arterial pressure (MAP) in anesthetized C57BL/6 mice. Ex vivo experiments using mouse mesenteric artery rings were conducted to validate the role of periplocymarin on blood vessels. However, periplocymarin failed to induce vasoconstriction directly, and had no effects on vasoconstriction induced by phenylephrine (Phe) and angiotensin II (Ang II). In addition, vasodilatation induced by acetylcholine (Ach) was insusceptible to periplocymarin. Echocardiography was used to evaluate the effects of periplocymarin on cardiac function. The results showed that the injection of periplocymarin significantly increase the ejection fraction (EF) in mice without changing the heart rate. In vitro studies using isolated neonatal rat ventricular myocytes (NRVMs) revealed that periplocymarin transiently increased the intracellular Ca²⁺ concentration observed by confocal microscope. But in Ca²⁺-free buffer, this phenomenon vanished. Besides, inhibition of sodium potassium-activated adenosine triphosphatase (Na⁺-K⁺-ATPase) by digoxin significantly suppressed the increase of MAP and EF in mice, and the influx of Ca²⁺ in cardiomyocytes, mediated by periplocymarin. Collectively, these findings demonstrated that periplocymarin increased the contractility of myocardium by promoting the Ca²⁺ influx of cardiomyocytes via targeting on Na⁺-K⁺-ATPase, which indirectly led to the instantaneous rise of blood pressure.

Background: Chronic glomerulonephritis is a prevalent renal disorder in clinical practice. A metabolic imbalance in copper, a vital trace element in the human body, is potentially linked to a myriad of diseases. Consequently, this study aimed to elucidate the role of copper metabolism in the onset and progression of chronic glomerulonephritis. Methods: Public microarray datasets gene expression omnibus series(GSE) 66494, GSE32591, and GSE116626 were procured from the gene expression omnibus database, encompassing clinical information on chronic glomerulonephritis and normal kidney tissues. Differential gene expression analysis was conducted, followed by the intersection of the differentially expressed genes from the 3 datasets with those related to copper metabolism. Furthermore, STRING and Cytoscape were used to construct a protein–protein interaction (PPI) network; PPI hub genes were identified via the cytoHubba plugin, and gene set enrichment analysis was performed. Tissues were subsequently collected from chronic glomerulonephritis patients at our hospital; hematoxylin and eosin, periodic acid-silver methenamine, and Masson staining were conducted; and transmission electron microscopy was performed. Pivotal genes were selected for real time quantitative polymerase chain reaction (PCR) (RT-qPCR) validation, and serum copper ion levels were measured. Additionally, CIBERSORT analysis was employed to assess immune cell infiltration and its correlation with pivotal genes. Results: Differential gene expression analysis revealed 25 differentially expressed copper metabolism-related genes (CMRGs), comprising 11 upregulated genes and 14 downregulated genes. PPI network construction and gene correlation analysis ultimately identified aconitase (ACO1) and superoxide dismutase (SOD2) as pivotal genes for RT-qPCR validation. The results demonstrated that ACO1 was expressed at low levels in lupus nephritis tissues, whereas SOD2 was highly expressed in immunoglobulin A nephropathy and diabetic nephropathy tissues. Moreover, immune infiltration analysis revealed that diverse immune cell types, including T cells, B cells, and macrophages, are intricately associated with the onset and progression of chronic glomerulonephritis in patient tissues. Differential and correlation analyses of CMRGs revealed that T cells, plasma cells, and monocyte-macrophages are involved in the biological processes of copper metabolism. Conclusion: This study suggests that the CMRGs ACO1 and SOD2 may modulate the inflammatory response and immune microenvironment in chronic glomerulonephritis by regulating immune cell activation and function, thereby facilitating disease progression.

Background: Intestinal ischemia–reperfusion injury (IIRI) is driven by oxidative stress, excessive inflammation, Endoplasmic Reticulum Stress (ERS), and disruption of the intestinal epithelial barrier. Corilagin, a natural ellagitannin with antioxidant and anti-inflammatory activities, has shown protective effects against IIRI; however, its cross-species and multitarget mechanisms remain unclear. Methods: This study investigated the protective mechanisms of Corilagin against IIRI by integrating Corilagin-related targets, IIRI-associated genes, and an ERS gene set. Differential expression analyses of GSE37013 and GSE96733 were combined with an ensemble machine-learning framework comprising 15 algorithms to prioritize core genes. Molecular docking and 100-ns all-atom molecular dynamics simulations using the AlphaFold-predicted full-length human NRF2 structure were performed to assess the predicted Corilagin-NRF2 interaction. A mouse IIRI model was used to validate selected molecular and histological findings by histology, RT-qPCR, Western blotting, and immunohistochemistry. In IEC-6 cells subjected to OGD/R, NRF2 immunofluorescence and siRNA-mediated knockdown of Nrf2 or Abcg2 were used to further evaluate the involvement of NRF2 and ABCG2 in Corilagin-mediated protection. Results: Intersection analysis identified 472 genes shared among Corilagin-related targets, IIRI-related genes, and ERS-associated genes. Enrichment analysis highlighted stress- and injury-related pathways, including PI3K-Akt signaling. Ensemble machine learning prioritized 12 core genes across human and mouse datasets, with ABCG2, IL10, and PRKCZ emerging as prominent candidates. Docking and 100-ns MD simulations supported persistent Corilagin retention within the predicted NRF2 binding region, while post-relaxation MM/GBSA analysis indicated favorable interaction energetics. In vivo , Corilagin pretreatment reduced inflammation and oxidative stress-related changes, restored apical ABCG2 and tight-junction protein expression, improved mucosal morphology, and altered CHOP expression. In IEC-6 cells, Nrf2 or Abcg2 knockdown weakened Corilagin-mediated preservation of Occludin, Claudin-1, and ZO-1 under OGD/R, supporting NRF2–ABCG2 involvement in this protective response. Conclusion: Corilagin may alleviate IIRI by modulating ERS-related molecular changes and preserving the intestinal epithelial barrier with NRF2 and ABCG2 contributing to a candidate regulatory module involved in this protective response. These findings provide computational and experimental support for further investigation of Corilagin as a potential intervention for IIRI and illustrate the value of combining multi-source data integration with ensemble machine learning to characterize the multitarget pharmacology of natural products.