Cryptotanshinone

Cryptotanshinone serves as the principal bioactive constituent of Salvia miltiorrhiza Bunge, possesses a wide range of pharmacological activities. Salvia miltiorrhiza Bunge, a long-standing therapeutic agent in traditional Chinese medicine (TCM) practice, is renowned for its efficacy in enhancing blood circulation and alleviating blood stasis and infarction, thereby treating cardiovascular and cerebrovascular diseases.

Platelet activation, when excessive or aberrant, poses a significant risk, catalyzing the onset of various thrombotic disorders. Thus, this investigation is meticulously designed to assess the antiplatelet pharmacological activity of cryptotanshinone, delving into its mechanisms of action that operate independently of the P2Y12 receptor.

We employed a combination of isolated human platelet functional analysis, network pharmacology, molecular docking, and animal experiments to explore the P2Y12 receptor-independent antiplatelet targets and the biological mechanisms by which cryptotanshinone improves thrombosis.

Utilizing the ADP-hydrolyzing enzyme apyrase, we isolated the direct effects of cryptotanshinone on platelet function. The findings reveal that cryptotanshinone can effectively inhibit platelet activation in a manner that is independent of the P2Y12 receptor, all the while maintaining normal tail bleeding times in murine models and not exacerbating mesenteric thrombosis. These effects appear to be mediated through intricate signaling pathways, including PI3K-AKT, MAPK, and STAT3.

This study compellingly confirms the capacity of cryptotanshinone to suppress platelet function independently of the P2Y12 receptor, establishing a robust theoretical foundation for innovative strategies in thrombosis prevention.

Previous studies have shown that testosterone activates the GPRC6A-Duox1 axis, resulting in the production of H₂O₂ which leads to the apoptosis of keratinocytes and ultimately hair loss. Here, we elucidated a molecular mechanism by which the non-genomic action of testosterone regulates cellular redox status in androgenetic alopecia (AGA). Building upon this molecular understanding, we conducted a high-throughput screening assay of Nox inhibitors from a natural compounds library. This screening identified diterpenoid compounds, specifically Tanshinone I, Tanshinone IIA, Tanshinone IIB, and Cryptotanshinone, derived from Salviae Miltiorrhizae Radix. The IC₅₀ values for Nox isozymes were found to be 2.6-12.9 μM for Tanshinone I, 1.9-7.2 μM for Tanshinone IIA, 5.2-11.9 μM for Tanshinone IIB, and 2.1-7.9 μM for Cryptotanshinone. Furthermore, 3D computational docking analysis confirmed the structural basis by which Tanshinone compounds inhibit Nox activity. These compounds were observed to substitute for NADPH at the π-π bond site between NADPH and FAD, leading to the suppression of Nox activity. Notably, Tanshinone I and Tanshinone IIA effectively inhibited Nox activity heightened by testosterone, consequently reducing the production of intracellular H₂O₂ and preventing cell apoptosis. In an animal study involving the application of testosterone to the back skin of 8-week-old C57BL/6J mice to inhibit hair growth, subsequent treatment with Tanshinone I or Tanshinone IIA alongside testosterone resulted in a substantial increase in hair follicle length compared to testosterone treatment alone. These findings underscore the potential efficacy of Tanshinone I and Tanshinone IIA as therapeutic agents for AGA by inhibiting Nox activity.