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DOI: 10.1055/a-0660-0441
Cryptotanshinone from Salvia miltiorrhiza Roots Reduces Cytokeratin CK1/10 Expression in Keratinocytes by Activation of Peptidyl-prolyl-cis-trans-isomerase FKBP1A
Publication History
received 13 April 2018
revised 02 July 2018
accepted 11 July 2018
Publication Date:
23 July 2018 (online)
Abstract
Cryptotanshinone (CTS) (1 µM) from the roots of Salvia miltiorrhiza exerts a strong influence on the terminal differentiation of human keratinocytes (HaCaT cell line, primary natural human keratinocytes) and downregulates the expression of differentiation-specific cytokeratins CK1 and CK10 on protein and gene level. Other differentiation specific proteins as involucrin, filaggrin, loricrin, and transglutaminase were not affected to a higher extent. CTS (1 µM) did not influence the cell viability and the proliferation of keratinocytes. Using a combination of drug affinity response target stability assay in combination with a proteomic approach and multivariate statistics for target elucidation, peptidyl-prolyl-cis-trans-isomerase FKBP1A (known target of inhibitors such as tacrolimus or rapamycin) was addressed as potential molecular target of CTS. The interaction of CTS with FKBP1A was additionally shown by thermal shift and enzymatic activity assays. Interestingly, CTS served as an activator of FKBP1A, which led to a reduced activity of the TGFβ receptor pathway and therefore to a diminished CK1 and CK10 expression. The combination of the FKBP1A activator CTS with the inhibitor tacrolimus neutralized the effects of both compounds. From these data, a potential dermatological use of CTS and CTS-containing plant extracts (e.g., hydroalcoholic extract from the roots of S. miltiorrhiza) for keratinopathic ichthyosis, a disease characterized by overexpression of CK1 and CK10, is discussed. This study displays an experimental strategy for combining phytochemical aspects on active natural products with systematic identification of molecular targets on gene, protein, and cell level.
Key words
CFKBP1A - cryptotanshinone - differentiation - keratinocytes - Salvia miltiorrhiza - Lamiaceae - TGFβ receptorSupporting Information
- Supporting Information
Influence of CTS at different concentrations on the cellular dehydrogenase activity of HaCaT keratinocytes (Fig. 1S). Influence of extract Ew and EE on the cellular dehydrogenase activity of HaCaT keratinocytes (Fig. 2S). Influence of CTS on the relative cell proliferation of NHEKs (Fig. 3S). Influence of CTS (1 µM) on the relative normalized expression of 14 genes in NHEK after 60 of incubation (Fig. 4S). Relative increase in cell layer of HaCaT cells within scratch assay over 24 h after treatment with CTS (1, 2 µM) and TGFβ (Fig. 5S). Biochemical functions of 14 genes, selected during differentiation gene expression array, and for which relevant fold changes were observed during cell contact with CTS (Table 1S). Overview of selected proteins, influenced by CTS in cell lysates obtained from HaCaT cells and with changed stability against proteolytic digest by thermolysin (Table 2S). Overview on the relative changes within the cell cycle of NHEK after incubation with CTS and TGFβ, related to the UCs (Table 3S).
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