Shikonin promotes hypertrophic scar repair by autophagy of hypertrophic scar-derived fibroblasts

Abstract

To investigate the Shikonin (SHI) induce autophagy of hypertrophic scar-derived fibroblasts (HSFs) and the mechanism of which in repairing hypertrophic scar. Methods: This study showed that SHI induced autophagy from HSFs and repaired skin scars through the AMPK/mTOR pathway. Alamar Blue and Sirius red were used to identify cell activity and collagen.Electron microscopy, label-free quantitative proteomic analysis, fluorescence and other methods were used to identify autophagy. The differences in the expression of autophagy and AMPK/mTOR pathway-related proteins after SHI treatment were quantitatively analyzed by Western blots. A quantitative real-time polymerase chain reaction assay was used to detect the expression of LC3, AMPK and ULK after adding chloroquine (CQ) autophagy inhibitor. Results: After treatment with SHI for 24 hours, it was found that the viability of HSFs was significantly reduced, the protein expression of LC3-II/LC3-I and Beclin1 increased, while the protein expression of P62 decreased. The expression of phosphorylated AMPK increased and expression of phosphorylated mTOR decreased. After the use of CQ, the cell autophagy caused by SHI was blocked. The key genes LC3 and P62 were then reexamined by immunohistochemistry using a porcine full-thickness burn hypertrophic scar model, and the results verified that SHI could induce autophagy in vivo. Conclusion: These findings suggested that SHI promoted autophagy of HSFs cells, and the potential mechanism may be related to the AMPK/mTOR signal pathway, which provided new insights for the treatment of hypertrophic scars.

Integrative Analysis of Lactylome and Proteome of Hypertrophic Scar To Identify Pathways or Proteins Associated with Disease Development

“…58 The promotion of autophagy in HSF cells by shikonin may be attributed to the AMPK/mTOR signal pathway, offering novel insights into hypertrophic scar treatment. 59 In this study, the integrative analysis of the lactylome and proteome revealed that hypertrophic scar tissues exhibited a significant upregulation of 14 lactylation sites. Our particular focus was on the lactylated GATM at K102.…”

“…It is well-established that hypoxia, leading to the induction of hypoxia-inducible factor-1 alpha (HIF-1alpha), contributes to scar formation during the healing of cutaneous wounds . The promotion of autophagy in HSF cells by shikonin may be attributed to the AMPK/mTOR signal pathway, offering novel insights into hypertrophic scar treatment …”

Pharmacotherapy for Keloids and Hypertrophic Scars

“…Shikonin has a role in inducing ROS, suppressing the release of exosomes, and inducing apoptosis [309]. Shikonin inhibits the expression of p63 (a type II integral membrane protein), cytokeratin 10, α-SMA, TGF-β1, and collagen I, which play important roles in HTS formation, suggesting that shikonin has potential utility as a novel scar therapy [310]. Also, shikonin promotes HTS repair via the autophagy of HTS-derived fibroblasts, where the potential mechanism may be related to the AMP-activated protein kinase/mammalian target of rapamycin (mTOR) signal pathway [311].…”

“…Orally administered saireito can reduce postoperative edema after blepharoptosis surgery by suppressing TGF-β1-induced Smad 2/3 phosphorylation [254,314]. Shikonin inhibits the expression of p63, cytokeratin 10, α-SMA, TGF-β1, and collagen I [310].…”