ETHNOPHARMACOLOGICAL RELEVANCE: Hongsheng Dan, a traditional Chinese medicine, has been utilized in China for centuries due to its therapeutic properties. Its primary active component, mercuric oxide (HgO), is known for its efficacy in extracting toxins, pus drainage, necrotic tissue removal, and tissue regeneration. Despite its longstanding clinical application, the precise therapeutic mechanisms and comprehensive safety profile of Hongsheng Dan warrant further scientific investigation.
AIMS OF THE STUDY: This study aims to elucidate the therapeutic mechanisms of Hongsheng Dan in the treatment of chronic, refractory skin ulcers and to evaluate its potential toxicity risks.
METHODS: A rat model of chronic refractory wound was successfully established to evaluate the therapeutic effects of Hongsheng Dan. Wound healing efficacy was systematically assessed through multiple approaches, including wound closure rate, histopathological examination, immunohistochemical and immunofluorescence staining, and Hyp content analysis. To elucidate the underlying mechanisms, qRT-PCR and WB analyses were performed. Additionally, H&E staining was conducted to examine pathological alterations in major organs, while AAS was employed to quantify mercury ion accumulation.
RESULTS: Hongsheng Dan demonstrated significant wound-healing efficacy in a rat model, with optimal therapeutic outcomes observed in the medium-dose treatment group. Histopathological analysis revealed substantial improvements in wound tissue architecture, characterized by diminished inflammatory cell infiltration and enhanced collagen deposition with robust granulation tissue formation. These morphological improvements were supported by immunohistochemical evidence showing significantly upregulated VEGF expression.At the molecular level, Hongsheng Dan treatment modulated key wound-healing mediators, suppressing IL-6 while promoting Hyp synthesis, thereby facilitating extracellular matrix remodeling. Gene expression analysis revealed coordinated regulation of critical healing pathways, with upregulation of VEGF, TGF-β1, and Smad3 mRNA, concurrent with downregulation of Notch1 signaling. WB analysis confirmed these findings at the protein level, showing increased VEGF and decreased Notch1 expression.Notably, while therapeutic doses of Hongsheng Dan caused no observable histopathological abnormalities in major organs, atomic absorption spectrometry detected significant mercury accumulation in renal tissue, suggesting potential nephrotoxic risk during extended therapeutic use.
CONCLUSION: This investigation adopted a dual-focused approach to evaluate both the therapeutic mechanisms and potential toxicity of Hongsheng Dan. Prior to the systematic research, we performed quality control to ascertain the mercury valence state and concentration. Following this, a chronic refractory wound model in SD rats was studied using an integrated methodological framework that combining histopathological, molecular biological, and toxicological analyses. Our findings demonstrated that Hongsheng Dan promotes chronic wound healing through multi-pathway synergistic mechanisms. Concurrent concomitant toxicity revealed mercury accumulation-related toxicity, providing crucial evidence for risk-benefit evaluation in clinical applications.
