Study on the potential medical beauty efficacy screening of acetyl hexapeptide-8 and the mechanism of regulating collagen regeneration

Acetyl hexapeptide-8 (AH-8) is a synthetic polypeptide that can promote collagen regeneration; however, the underlying mechanism of this promoting effect and other potential medical beauty efficacy of AH-8 remain unknown. This study aims to explore the effect of AH-8 in promoting collagen regeneration and its underlying mechanism. In this study, various zebrafish efficacy evaluation models were constructed to systematically evaluate the potential medical efficacy of AH-8. At the same time, the promoting effect and mechanism of AH-8 on collagen regeneration were explored from the messenger ribonucleic acid (mRNA) and protein level, respectively. AH-8 promoted the expression of collagen and elastin genes, reduced locomotor activity in zebrafish under both basal conditions and in chemically-induced behavioral models, and decreased the expression of inflammatory factors. These findings suggest that AH-8 has the potential to regulate collagen regeneration, alleviate neural excitation, reduce stinging sensations, and exert soothing and anti-inflammatory effects. RNA sequencing analysis via high-throughput sequencing revealed that differentially expressed genes between the model group and the AH-8-treated group were enriched in pathways related to organismal aging regulation (e.g., the TGF- β signaling pathway), and these genes were upregulated after AH-8 treatment. Integrated data from transcriptomics, qPCR, ELISA, and CRISPR-dCas9-mediated knockdown consistently demonstrated that AH-8 concentration-dependently upregulated the expression of collagen genes ( col1a1a, col1a1b, col1a2 ) and the elastin gene ( eln1 ). Corresponding concentration-dependent changes in key pathway genes ( tgfb3, tgfrb1b, smad3b, smad4a/b ) aligned with the transcriptome sequencing results. AH-8 exhibits anti-wrinkle, nerve-soothing, sting-relieving, mild and non-irritating, skin-brightening, moisturizing, immediate anti-wrinkle, soothing, and anti-inflammatory efficacy. AH-8 probably promotes collagen regeneration by regulating key genes, such as col1a1a, col1a1b, col1a2, eln1, and smad4b, within the TGF - β /Smad2/3 signaling pathway, and by influencing the expression of related factors. • AH-8 dermal absorption in zebrafish confirmed by fluorescence, boosting collagen, locomotion, and easing tingling. • AH-8 exhibits anti-inflammatory activity by reducing neutrophil migration/aggregation in a concentration-dependent manner. • AH-8 suppresses inflammation and enhances collagen regeneration via TGF- β /Smad2/3 pathway. • This study provides a theoretical foundation for the application of AH-8 as an anti-aging component.