Journal: Redox Biology

Article Title: Cannabidiol induces antioxidant pathways in keratinocytes by targeting BACH1

doi: 10.1016/j.redox.2019.101321

Figure Lengend Snippet: Multi-omic analysis of the response of keratinocytes to CBD. The transcriptomic and proteomic profiling of RNA and protein samples was carried out using RNA-Seq and LC-MS/MS, respectively. (A, B) Volcano plots showing the magnitude (log2 fold change) and significance (-log10 p value) of the changes in the transcriptomic and proteomic comparisons of CBD treated keratinocytes versus controls (n = 3 for RNA-Seq and n = 4 for proteomics). Every point represents a gene/protein and the colour indicates those surpassing the cut-off of an adjusted P value < 0.05 and an absolute fold change >2 (for genes) or > 1.5 (for proteins). For RNA-Seq, a small value (1e-300) was added to p values in order to avoid logarithms of zero at plotting. (C) Upset plot indicating the overlap between the sets of up or down regulated genes and proteins as a bar plot over a coincidence histogram. (D) Over-representation analysis results. The dot plot indicates with a point the significant over-representation of a given term, transcription factor or pathway in a group of up or down regulated genes/proteins (Fisher Exact Test adjusted P < 0.1). While the colour indicates the adjusted P value of the enrichment, the size of the point represents the enrichR combined score.

Article Snippet: Normal human epidermal keratinocytes (NHEK) and Keratinocyte growth medium were purchased to Innoprot SL (Biscaia, Spain).

Techniques: RNA Sequencing Assay, Liquid Chromatography with Mass Spectroscopy

Journal: Redox Biology

Article Title: Cannabidiol induces antioxidant pathways in keratinocytes by targeting BACH1

doi: 10.1016/j.redox.2019.101321

Figure Lengend Snippet: Validation of the NRF2 pathway as a target of CBD in keratinocytes. (A) Gene set enrichment analysis plot for the NRF2 signalling pathway transcriptomic changes. Black lines indicate the position of NRF2 genes in the pre-ranked gene list and the green line indicates the running enrichment score. (B) Magnitude of the changes for significantly up-regulated genes and proteins selected using the previously mentioned cut-offs in the CBD versus control comparison . C) Primary human keratinocytes were incubated with either DMSO or CBD (10 μM) for 24 h. The mRNA levels for HMOX1 ( upper panel ) and SQSTM1 ( p62) ( lower panel ) were quantified using real-time PCR. The data were normalized using HPRT1 as an internal control. Data represent means ± SD (n = 3) and are expressed relative to the DMSO sample. ***P ≤ 0.001 . D) HaCaT cells were incubated with either DMSO or increasing concentration of CBD for 16 h. The mRNA levels for HMOX1 ( upper panel ) and SQSTM1 ( p62) ( lower panel ) were quantified using real-time PCR as previously indicated (n = 3) *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001. E) HaCaT-ARE-Luc cells were treated with either SFN or CBD at the indicated concentrations for 6 h. Luciferase activity was measured in the cell lysates and expressed as RLU (x 10 4 ). Data represent means ± SD (n = 4) and are expressed relative to untreated cells. **P ≤ 0.01, ***P ≤ 0.001.

Article Snippet: Normal human epidermal keratinocytes (NHEK) and Keratinocyte growth medium were purchased to Innoprot SL (Biscaia, Spain).

Techniques: Incubation, Real-time Polymerase Chain Reaction, Concentration Assay, Luciferase, Activity Assay

Journal: Redox Biology

Article Title: Cannabidiol induces antioxidant pathways in keratinocytes by targeting BACH1

doi: 10.1016/j.redox.2019.101321

Figure Lengend Snippet: CBD treatment increases the keratinocyte layer in the epidermis and the expression of HMOX1. (A) Haematoxylin-eosin staining of 5 μm paraffin-embedded sections were analysed by bright field microscopy. (B) Representative images of HMOX1 immunohistochemistry from mouse skin after 5 days of treatment with vehicle, CBD 0.1% or 1% (C) Quantification of HO-1 stained area in mouse epidermis. *P < 0.05, ***P < 0.001 compared with control. Scale bars: 200 (left) and 100 μm (right). n = 15 for vehicle treated samples and n = 12 for CBD treated samples.

Article Snippet: Normal human epidermal keratinocytes (NHEK) and Keratinocyte growth medium were purchased to Innoprot SL (Biscaia, Spain).

Techniques: Expressing, Staining, Microscopy, Immunohistochemistry